Abstract: Polynucleotide sequences are provided for the diagnosis of the presence of retroviral infection in a human host associated with lymphadenopathy syndrome and/or acquired immune deficiency syndrome, for expression of polypeptides and use of the polypeptides to prepare antibodies, where both the polypeptides and antibodies may be employed as diagnostic reagents or in therapy, e.g., vaccines and passive immunization. The sequences provide detection of the viral infectious agents associated with the indicated syndromes and can be used for expression of antigenic polypeptides.

Abstract: This invention provides isolated nucleic acid molecules encoding human 5-HT.sub.1D receptors, isolated proteins which are human 5-HT.sub.1D receptors, vectors comprising isolated nucleic acid molecules encoding human 5-HT.sub.1D receptors, mammalian cells comprising such vectors, antibodies directed to the human 5-HT.sub.1D receptors, nucleic acid probes useful for detecting nucleic acid encoding human 5-HT.sub.1D receptors, antisense oligonucleotides complementary to any sequences of a nucleic acid molecule which encodes a human 5-HT.sub.1D receptor, pharmaceutical compounds related to human 5-HT.sub.1D receptors, and nonhuman transgenic animals which express DNA a normal or a mutant human 5-HT.sub.1D receptor. This invention further provides methods for determining ligand binding, detecting expression, drug screening, and treatment involving the human 5-HT.sub.1D receptor.

Abstract: Basement membrane proteins are isolated as functioning proteins in relatively large amounts from human or animal tissues in aqueous solution in the presence of a chelating agent. It is possible to use these proteins to obtain highly specific antibodies which are used for the immunological determination of these proteins.

Abstract: An enzyme complex having collenagenolytic activity, comprising a mixture of collegenolytic proteases from crabs is disclosed. The proteases of the complex have the ability to cleave bovine lens capsule collagen type IV, have molecular weights from about 23,000 to 36,000 daltons, and possess a collagenolytic activity of more than about 3.300 Mandl units/mg at a pH of 7.5. The complex is isolated in aqueous solutions without the use of organic solvents from the hepatopancreas of crabs. The crabs from which the complex can be isolated are of the paralithodes and chinocetes species of crabs.

Abstract: An assay method is provided to quickly detect the presence of Listeria strains in samples, characterized by the use of antibodies to selectively capture the peptidoglycan and teichoic acid components of the listeriae bacterial cell wall.

Abstract: The present invention relates to (1) an enzyme-linked immunosorbent assay (ELISA) for detection in milk of antibodies of any isotype which are specific for Staphylococcus aureus proteins in molecular weights ranging from 14,000 to 26,000 daltons, (2) a process for production and purification of the proteins, (3) a method of performing the ELISA utilizing the proteins (4) use of the ELISA for detection of intramammary infection by S. aureus, (5) preparation of monoclonal and polyclonal antibodies against the selected fractions, (6) use of such antibodies of purification of infection specfic antigens, and (7) use of the antibodies in an ELISA for antibodies produced in an individual in response to infection by S. aureus.

Abstract: A kit for testing materno-fetal immunoincompatibility in women, particularly in women whose fetuses are in danger, comprises two containers containing an identical amoung of HLA-D antigen-containing material and one container containing male serum as control. In the test, maternal serum of the patient is added to one container of antigen while the control serum is mixed with the other container of antigen. The amount of expressed HLA-D antigen remaining in each case is established by conventional HLA-D directed monoclonal antibody immunoassay techniques such as conventional radio-immunoassay. The maternal serum assay divided by the control serum assay gives an immunoincompatibility quotient (IQ). The IQ is from 40 to 50% in healthy pregnant women and is about 70% in pathological cases. The test provides reliable results since the antigenic portion is non-living and therefore does not offer the inconstances of living test reagents.

Abstract: The present invention relates to methods and compositons for isolation of nucleic acids from cells. In particular aspects, this invention relates to the use of chaotropic compositions, such as guanidine hydrochloride or guanidinium isothiocyanate, in combination with polyanionic compositions, such as those containing sulfated polysaccharides (i.e., heparin or heparitin sulfate), for the isolation of nucleic acids (RNA or DNA). This method involves disrupting and lysing cells using a nucleic acid releasing composition containing a chaotropic component for the release of nucleic acids from the cell (guanidine hydrochloride or guanidinium isothiocyanate). The released nucleic acids are collected by ethanol precipitation and resuspended before exposure to a polyanion-containing protein dissociating composition which promotes the dissociation of nucleic acid associated proteins from the resuspended nucleic acids.

Abstract: Peptides and proteins related to an epitope comprising an outer membrane protein of Haemophilus influenzae are described. The peptides and proteins can be prepared by methods including novel and improved methods of puiification from H. influenzae cultures, and by recombinant DNA and chemical synthetic techniques. Additionally, recombinant vectors containing nucleotide sequences encoding PBOMP-1 and PBOMP-2 related peptides and proteins are also described. Recombinant vectors include plasmid DNA and viral DNA such as human viruses, animal viruses, insect viruses and bacteriophages that direct the expression of the PBOMP-1 and PBOMP-2 related peptides and proteins in appropriate host cells. The peptides, proteins and viruses both "live" and "inactivated" are used as immunogens in vaccine formulations to protect against H. influenzae infections. The peptides and proteins are also used as reagents in immunoassays as well as to prepare immunoglobulins for passive immunization.

Abstract: The subject invention concerns a novel in vitro process for identifying and quantifying native antigens on potentially pathogenic group B streptococci bacteria present in a clinical specimen. The invention process is made possible by the discovery of novel bacterial markers denoted .gamma. and .delta. epitopes which are expressed by a variety of group B streptococcal strains.

Abstract: A substantially pure capsular exopolysaccharide adhesin of coagulase-negative staphyhlococcal strains, and a general method to prepare such adhesins, are described. Vaccines composed of such adhesins, and uses of such adhesins to produce polyclonal and monoclonal antibodies against such adhesins, are also disclosed. The adhesins are useful in coating polymeric medical materials to prevent colonization by coagulase-negative staphylococcal strains, and as a probe in selecting desirable polymeric medical materials. Such adhesin antibodies are useful in vivo to prevent infection by nosocomial coagulase-negative staphylococcal strains, in assays for the detection of such bacteria, in assays for the estimation of such adhesins in complex mixtures, and as an affinity chromatography matrix.

Abstract: This invention relates to an improvement in nucleic acid hybridization technology. Nucleic acids bind to complementary partners in a predictable manner such that the detection of complementary partners is possible. The acceleration of the binding process is desired objective and will find broad application in a variety of industrial, medical, and research uses. In particular this invention relates to the acceleration of nucleic acid hybridization by heterogeneous nuclear ribonucleoproteins [hnRNPs].

Abstract: Composition for the prevention and treatment of autoimmune diseases are provided which comprise as an active ingredient membrane material shed from autoimmune T lymphocytes, or activated T lymphocytes which are treated by a pressure application and releases process. There is also provided processes for obtaining such active materials and for preparing pharmaceutical compositions containing them.

Abstract: The present invention is directed to a novel method, composition and device for removing dissolved oxygen from solutions containing alcohols and/or acids. By removing oxygen from various products, the present invention is an effective antioxidant for beverages and food products, as well as for industrial and commercial solutions containing alcohols and/or acids.

Abstract: A process is disclosed for recovering a cell wall protein, particularly a Fc-receptor, from streptococcal bacteria. In the process, streptococcal bacteria are treated with at least one proteolytic enzyme to solubilize the cell wall protein. A process is also disclosed for recovering Fc-receptor type III (protein G) having selective IgG-binding capability (i.e., no albumin-binding capability) from streptococcal bacteria by pretreating these bacteria with enzymes prior to said enzymatic solubilization.

Abstract: An improved hollow fiber bioreactor system is disclosed that includes an arrangement for removing the plasticizer that is typically found within the hollow fibers of hollow fiber cartridges so as to prevent this material from contaminating the nutrient fluid. The plasticizer is removed without jeopardizing sterility of the bioreactor.

Abstract: An immunogenicity increasing carrier substance for immunogenic determinants is composed of particles derived from gram-negative cells substantially devoid or depleted of their natural immunogenic determinants, more particularly prepared by stripping the O-antigen and core region off the lipid A by chemical cleavage (acid hydrolysis). These socalled "naked bacteria" are chemically treated with a linking reagent to provide covalently bonded intermediate linking moieties while preserving the adjuvant effect of the particles. The linking moieties have functional groups. These serve for the covalent bonding of selected haptens or antigens comprising the desired immunogenic determinants against which antibodies are to be elicited in living antibody-producing cells.

Abstract: A vessel containing a polymeric acid is used in a method and test kit for extracting a bacterial (e.g., streptococcal) antigen from a test sample, for example, preliminary to an immunoassay. To extract the antigen from bacteria in the sample, a precursor reagent is applied to the vessel acid and incubated with the sample. The kit includes a vial containing the acid and another vial containing the precursor. The kit is produced by including the acid polymer in the vial or vessel, e.g., as a pellet or coating.

Abstract: The present invention relates to (1) an enzyme-linked immunosorbent assay (ELISA) for detection in milk of antibodies of any isotype which are specific for Staphylococcus aureus proteins in molecular weights ranging from 18,000 to 26,000 daltons, (2) a process for production and purification of said proteins, (3) a method of performing said ELISA utilizing said proteins and (4) use of said ELISA for detection of intramammary infection by S. aureus.

Abstract: A process for the separation from other cellular materials of heat agglomeration resistant water soluble nitrogen containing organic compounds such as plasmids, RNA's, mitochondrial DNA's, viral DNA's, chloroplast DNA's, other episomal DNA's and certain proteins. The process comprises heating cellular materials in a solution of lysing agent to lyse the desired cells and to agglomerate water soluble nitrogen containing compounds such as certain chromosomal DNA's which are not resistant to agglomeration; centrifuging the resulting product to remove water soluble agglomerated materials; separating the supernatant liquid and precipitating the water soluble agglomeration resistant organic compounds with a water soluble precipitant. The process also includes separating the agglomeration resistant water soluble nitrogen containing compounds from each other by means of exclusion chromotography.

Abstract: A method of testing a fluid sample for the presence of antibodies against a micro-organism, which comprises contacting the sample with fixed cells or fragments of cells infected with the micro-organism, and determining the presence of antibody bound to the cell-associated antigens, in which the determination is by virtue of a color change visible to the naked-eye at the site of the antibodies. For use in a testing method of this type, a test component comprises upper and lower layers, in which the upper layer has an array of apertures through which discrete areas on the lower layer are exposed, and in which the lower layer carries, in some at least of the discrete areas, fixed cells or cell fragments infected by a micro-organism.

Abstract: An antigen which, as its major immunizing component, comprises a determinant of an adhesin polypeptide or an immunogenically active subsequence thereof or a precursor therefor which is convertible to an immunogenically active form, antibodies against which determinant react with the adhesion polypeptide produced by pathogenic adhesin-forming bacteria which adhere to mammalian tissue, antibodies against such antigen, and DNA expressing, as a principal gene product thereof, such antigen.

Abstract: A 60-like antigens isolated in a one-step reaction from the bacterial cytoplasma of Mycobacteria are useful for the production of diagnostic tests.

Abstract: Substances that are toxic to yeast and which cause cessation of fermentation during alcoholic fermentation are adsorbed by microorganism cell walls added to a medium being fermented. The cell walls are from a gram-positive microorganism such as yeast, and are obtained by boiling or autolysis of the microorganism followed by washing material recovered. The cell walls can be added before or during fermentation, and may be added to a previously fermented medium followed by inoculating with new yeast. The toxic substances may be certain fatty acids and their ethyl esters, pesticide residues and substances secreted by certain microorganisms. Preferably, the cell walls are added when making wine, and the medium may contain Botrytis cinerea.

Abstract: An improved assay for measuring the quantity of Beta-lactam antibiotics, particularly valuable for measuring penicillin in milk. The assay includes the covalent-attachment of a penicillin to a penicillin-binding-protein bound to a matrix, the highly selective attachment of an antibody to such a conjugate, and the detection and measurement of the quantity of resulting antibody-complex material as a direct measure of the amount of penicillin in the milk being tested.

Abstract: An improved assay for an ATL virus antibody in a specimen, in which an ATL virus antigen is added to the test specimen and used as a comparative specimen. This improved method selectively assays ATL virus antibodies, and, thus, is useful in preventing and treating adult T cell leukemia.

Abstract: The present invention is a method for expressing functional polypeptides in Streptomyces using a recombinant DNA expression vector comprising a novel transcriptional- and translational- activating sequence. The novel activating sequence can be synthesized by conventional methods and used in Streptomyces expression vectors. One such vector, plasmid pFJ350, expresses and confers hygromycin resistance in Streptomyces host cells.

Abstract: Industrially useful method for the purification of HBs antigen produced by a recombinant organism being capable of producing HBs antigen which is prepared by means of DNA recombination technique, which comprises subjecting an HBs antigen-containing material produced by a recombinant organism to an adsorption chromatography with a silica, optionally followed by a gel filtration and further an adsorption chromatography with a hydroxyapatite, and then eluting the HBs antigen, preferably, with a buffer having a pH 9 or more which is incorporated with urea. The purification method can give a highly pure HBs antigen suitable for the preparation of hepatitis B vaccine in a large scale from recombinant organisms prepared by DNA recombination technique.

Abstract: There are provided a crystalline and single rod products derivable from the Pili of Type 1 and Type 2 Neisseria gonorrhoese organisms. There are provided methods of growing said organisms to produce the maximum yield of Pili and procedures for purifying said Pili to produce said crystalline material. There are further provided methods of utilizing said Pili to determine the presence, in a system infectable by N. gonorrhoese organisms, of antibodies to the Pili of said organisms, and methods of serotyping said Pili. There is also provided a mode of utilizing said crystalline material to provide a substantial degree of immunization infection by N. gonorrhoese in mammalian systems susceptible to such infection.

Abstract: A method for detecting or removing a substance in a medium is presented. Magnetic material, particularly magnetic bacteria or magnetic particles contained therein, are treated to render them receptive to binding or attachment to the substance sought to be detected or removed. Following binding or attachment to the treated magnetic material, the medium is subjected to a magnetic field, which results in removal of the magnetic material and the substance bound or attached thereto.

Abstract: A method of specimen treatment preparatory to conducting an immunoassay is disclosed whereby a microbial protein is solubilized by a detergent at elevated temperatures and in the presence of an alkali or alkaline earth metal ion. At elevated temperatures, the detergent is soluble. However, at lower temperatures, the presence of the metal ion renders the detergent insoluble so that it is prevented from interacting in the immunoassay procedure. A specific application is in the solubilization of the principal outer membrane protein of Chlamydia trachomatis.

Abstract: In the colony hybridization technique, bacterial colonies are replica-plated onto filter paper, and their DNA is hybridized with labeled DNA containing a specific sequence. A one-hundred fold increase in sensitivity is obtained by subjecting the colonies to a stream of steam in the presence of alkali for about three minutes prior to hybridization.

Abstract: A reactor system and method for synthesizing or degrading polynucleotides and other linear polymers includes a tubular reactor connected to a reagent manifold. The polynucleotide is immobilized on a loosely packed solid-phase support material in the tubular reactor, and reagents are sequentially introduced into the tubular reactor. After each reagent is introduced, the tubular reactor is isolated from the reagent manifold and the reagent agitated by alternately pressurizing the opposite ends of the tubular reactor. The method provides rapid and efficient synthesis of polynucleotides. By connecting two or more tubular reactors to the reagent manifold, a plurality of polynucleotides having different sequences may be synthesized simultaneously.

Abstract: There is disclosed a filter comprising fibrin in gel form, the gel having substantially uniform pore sizes, and the filter comprising means for retaining the shape of at least one surface of the gel against deformation when contacted by a flowing medium.

Abstract: There is disclosed a filter comprising fibrin in gel form, the gel having substantially uniform pore sizes, and the filter comprising means for retaining the shape of at least one surface of the gel against deformation when contacted by a flowing medium.

Abstract: Gene products of plasmid DNA, such as proteins, are prepared in high yields by cultivating bacteria carrying a plasmid which shows a controlled constant plasmid copy number at one temperture and a much higher or totally uncontrolled copy number at a different temperature. The plasmid may be prepared by recombinant DNA technique using a cloning vector showing the temperature dependent plasmid copy number pattern.

Abstract: A reactor system and method for synthesizing or degrading polynucleotides and other linear polymers includes a tubular reactor connected to a reagent manifold. The polynucleotide is immobilized on a loosely packed solid-phase support material in the tubular reactor, and reagents are sequentially introduced into the tubular reactor. After each reagent is introduced, the tubular reactor is isolated from the reagent manifold and the reagent agitated by alternately pressurizing the opposite ends of the tubular reactor. The method provides rapid and efficient synthesis of polynucleotides.

Abstract: A material and method for promoting the growth of anaerobic bacteria which includes a nutrient media containing a hydrogen donor and sterile membrane fragments of bacteria having an electron transfer system which reduces oxygen to water. Dissolved oxygen in the medium is removed by adding the sterile membrane fragments to the nutrient medium and holding the medium at a temperature of about 10.degree. to about 60.degree. C. until the dissolved oxygen is removed.

Abstract: Method and compositions are provided for replication and expression of exogenous genes in microorganisms. Plasmids or virus DNA are cleaved to provide linear DNA having ligatable termini, which are bound to a gene having complementary termini, to provide a biologically functional replicon with a desired phenotypical property. The replicon is inserted into a microorganism cell by transformation. Isolation of the transformants provides cells for replication and expression of the DNA molecules present in the modified plasmid. The method provides a convenient and efficient way to introduce genetic capability into microorganisms for the production of nucleic acids are proteins, such as medically or commercially useful enzymes, which may have direct usefulness, or may find expression in the production of drugs, such as hormones, antibiotics, or the like, fixation of nitrogen, fermentation, utilization of specific feedstocks, or the like.

Abstract: A method of inserting deoxyribonucleic acid or fragments thereof into a living cell, which comprises; encapsulating the DNA or fragment in a lipid vesicle and bringing the vesicle in contact with said cell, whereby insertion occurs.

Abstract: A biologically pure culture of a bacterium of the genus Aquaspirillum, designated MS-1, has been found to contain chains (so-called magnetosomes) of single domain magnetite particles. The magnetite particles are roughly cubic and are about 500 A on a side and each of the chains contains approximately 290 magnetite particles. These magnetite particles can be recovered from the bacterium and usefully employed in magnetic recording devices and the like.

Abstract: PhoA mutant E. coli SB44, prepared by mutation of E. coli HB101 with an N-nitroso compound, can be used to identify and isolate recombinant plasmids into which a phoA gene has been incorporated. These plasmids can be used to transform bacteria which can be cloned and incubated to provide alkaline phosphatase in high yield. Moreover, these plasmid vectors can be modified in various ways so that the N-terminal amino acid sequence of phoA is followed in reading phase by the DNA coding for some other protein. In turn, these new plasmids can be used to transform bacteria which can be cloned and incubated to produce fusion proteins comprising the desired other protein in high yield and outside of the cell membrane in the periplasmatic space.

Abstract: An apparatus for the stepwise synthesis of polynucleotides in which the polynucleotide chains are extended in stepwise fashion from a modified form of polymer support to which the first unit is linked comprises a reaction column containing the polymer supported product and acting as the reaction vessel, and a series of reaction bottles all connected to the reaction column by means of a fluid flow conduit to which the vessels make connection via two-way valves arranged in series. The farthest upstream vessel of the series contains reaction solvent, used for washing purposes and the like. The most downstream of the reaction vessels contain nucleotide reagents.

Abstract: DNA comprising the naturally occurring nucleotide sequence coding for amino acids 44-90 of .beta.-lipotropin and including the entire coding region for .beta.-endorphin with the exception of the C-terminal glutamine was modified, transferred to an expression transfer vector, and expressed as a fusion protein. The fusion protein was further modified in vitro to yield mature .beta.-endorphin. .beta.-endorphin was purified from a bacterial lysate. The structure and biological activity of the resulting product was proven by immunological assay, and by two independent assays designed to demonstrate biological activity.

Abstract: Novel chemical compounds, recombinant plasmids pUC1019 and pUC-1020, which are obtained by covalent linkage of ca. 4.2 kb BclI restriction endonuclease fragment of the Streptomyces espinosus plasmid pUC6 into the BamHI endonuclease site of the E. coli plasmid pBR322. Plasmid pUC1024 is obtained by restructuring plasmid pUC1019. These plasmids are useful as cloning vehicles in recombinant DNA work. For example, using DNA methodology, a desired gene, for example, the insulin gene, can be inserted into the plasmids and the resulting plasmids can then be transformed into a suitable host microbe which, upon culturing, produces the desired insulin.

Abstract: An E. coli strain producing metabolically active but non-reproductive, anucleated live cells, which contain the K99 surface antigen, is obtained by transferring the K99 plasmid from a K99+ entero pathogenic E. coli strain into an E. coli strain which generates progeny of cells that lack the ability to multiply. From the resulting E. coli strain a "live" vaccine is prepared. Separation and purification of the anucleated live cells from growing cells is the basis for the "live" and chemically and/or physically unmodified vaccine. The vaccine induces the production of antibodies against growing and infective enteropathogenic K99+ E. coli in cattle and is, thus effective against coliform enteritis.

Abstract: A method is disclosed for measuring the concentration of nucleotides and the number of viable cells in a biological sample containing somatic and/or microbial cells by treatment of a sample of cells with surface active agents to selectively release the nucleotides without rupture of the cell membrane or walls, and measuring by means of a bioluminescent assay technique the concentration of nucleotides selectively released and thereby determining the number of viable cells in the sample.

Abstract: A method and a DNA linker are described for synthesizing relatively long double-stranded deoxyribonucleic acid sequences of defined composition. Short complementary single strand segments of oligonucleotides comprising part of the full sequence desired are synthesized using known procedures. Overlapping single strand segments are annealed forming double-stranded fragments which are inserted in cloning vectors and cloned in an appropriate host, both purifying the DNA fragments and amplifying the amount thereof. An adjacent fragment is then similarly synthesized in quantity and such fragments are inserted adjacent to the first synthetic introduced fragments in the cloning vectors, followed by cloning in an appropriate host. The procedure continues until the entire desired sequence has been formed, at which time it may be excised or cloned directly in the vectors upon which it was made.

Abstract: A method for constructing strains which produce aminoacids comprising combining of a DNA chromosome fragment of a donor microorganism containing genes controlling the synthesis of a selected aminoacid and having a mutation destroying the negative regulation of the synthesis of this aminoacid with a vector DNA molecule to form a hybrid DNA molecule. Use is made of a vector DNA molecule capable of providing amplification of the hybrid DNA molecule. The resulting hybrid DNA molecule is used for transforming cells of the recipient strain having the mutation blocking the synthesis of the selected aminoacid in this strain and the mutation partly blocking the related step of metabolism of this aminoacid to yield the strain capable of increased productivity of the selected aminoacid.

Abstract: A novel chemical compound, essentially pure plasmid pUC6, which is obtainable from a biologically pure culture of the microorganism Streptomyces espinosus biotype 23724a, NRRL 11439. The pUC6 plasmid is useful as a cloning vehicle in recombinant DNA work. For example, using DNA methodology, a desired gene, for example, the insulin gene, can be inserted into pUC6 and the resulting plasmid can then be transformed into a suitable host microbe which, upon culturing, produces the desired insulin.