Abstract: A method for culturing Langerhans islets to obtain an amount sufficient for transplant and autotransplant is disclosed. The islets are cultured in a culture serum (rat/human) medium which is supplemented with radical scavengers, growth factors, a matrix material, nerve growth factor, cell migrating/scattering factors and anti-integrin &bgr;1 antibody at proper the time during the culturing process. The medium is supplemented with radical scavengers and growth factors for the first time and then further supplemented with matrix material, radical scavengers, nerve growth factor and the growth factors around 12-24 hours after culturing. Thereafter, the medium is supplemented with growth factors, cell migrating/scattering factors and anti-integrin &bgr;1 antibody at 4-5 days into the culturing process. The culturing process is conducted for an extended period of time, so that any latent red blood cells are eliminated from the islet culture.

Abstract: The invention relates to the use of a secretor variant cell-line expressing the alpha moiety of human IgE binding protein to determine the allergic status of a given individual. Moreover, the cell-line is also used to provide an assay system for determining the allergenicity of substances and for subsequently providing therapeutic compostions which render said substances ineffective. In addition, the invention also relates to the use of said cell-line to determine the IgE independent irritancy of substances and compositions effective for attenuating the effects of said substances.

Abstract: The present invention provides an isolated population of non-mouse embryonic stem (ES) cells and methods of obtaining these ES cells. In one aspect, the target ES cells are obtained by co-culturing embryo cells from a target animal with non-target ES cells, such as mouse ES cells. In one embodiment, rat ES cells are isolated from the co-culture using positive or negative selectable markers.

Abstract: The orphan seven transmembrane domain receptor, APJ, can function as a coreceptor for cellular infection by the HIV virus. The establishment of cell lines that coexpress CD4 and APJ provide valuable tools for continuing research on HIV infection and the development of anti-HIV therapeutics.

Abstract: A polynucleotide encoding a secreted form of wild type Renilla luciferase. Also provided is a polynucleotide encoding a secreted modified form of wild type Renilla luciferase. Additionally, the polypeptides encoded by the polynucleotides of the present invention and uses of the polynucleotides and polypeptides of the present invention in biological assays. Also, a stable mammalian packaging cell line which produces retroviruses carrying a polynucleotide encoding a secreted Renilla luciferase.

Abstract: Rattus norvegicus AXOR12-RAT polypeptides and polynucleotides and method for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for screening for compounds that either agonize or antagonize Rattus norvegicus AXOR12-RAT. Such compounds are expected to be useful in treatment of human diseases, including, but not limited to: infections such as bacterial, fungal, protozoan and viral infections, particularly infections caused by HIV-1 or HIV-2; pain; cancers; diabetes, obesity; anorexia; bulimia; asthma; Parkinson's disease; acute heart failure; hypotension; hypertension; urinary retention; osteoporosis; angina pectoris; myocardial infarction; stroke; ulcers; asthma; allergies; benign prostatic hypertrophy; migraine; vomiting; psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, depression, delirium, dementia, and severe mental retardation; and dyskinesias, such as Huntington's disease or Gilles dela Tourett's syndrome.

Abstract: The JeT promoter is a recombinant promoter with transcriptional activity comparable to a number of strong mammalian promoters. The promoter consists of five key elements: (1) a TATA box; (2) a transcription initiation site (Inr); (3) a CAT consensus sequence in conjunction with (4) a CArG element and finally, (5) four Sp1 transcription binding sites (GGGCGG) arranged in two tandems.

Abstract: The present invention provides:

Abstract: The present invention features transgenic rodent models for HIV, wherein the transgenic rodent or transgenic rodent cell has incorporated into its genome genes encoding a human CD4 receptor and a human chemokine receptor. In particular the invention relates to transgenic rats, or mice characterized by being susceptible to infection with HIV, capable of expressing HIV structural genes, or HIV replication. The transgenic rodent or rodent cell of this invention is useful for studying the molecular basis of HIV infection, replication and pathogenicity, as well as for the testing of agents for new antiviral or vaccine strategies.

Abstract: rDNA promoter constructs useful in plasmids and vectors directing transcription of RNAs in a Pol I-specific manner constructed of four elements in a serial array: a first nucleotide sequence, capable of hybridizing under stringent conditions to an rDNA promoter element; a second nucleotide sequence, capable of hybridizing under stringent conditions to an internal ribosome entry signal (IRES); a third nucleotide sequence containing a coding region interest; a fourth nucleotide sequence containing a polyadenylation (polyA) signal sequence; a fifth rDNA enhancer element may be positioned upstream of the serial array. Also recombinant permissive cells and genetically engineered stable cell lines that contain the subject constructs.

Abstract: Multipotent neuroepithelial stem cells and lineage-restricted oligodendrocyte-astrocyte precursor cells are described. The neuroepithelial stem cells are capable of self-renewal and of differentiation into neurons, astrocytes, and oligodendrocytes. The oligodendrocyte-astrocyte precursor cells are derived from neuroepithelial stem cells, are capable of self-renewal, and can differentiate into oligodendrocytes and astrocytes, but not neurons. Methods of generating, isolating, and culturing such neuroepithelial stem cells and oligodendrocyte-astrocyte precursor cells are also disclosed.

Abstract: The transgenic animal of this invention can be used as an animal model for renal disease, bone disease, joint disease, pulmonary disease, hyperlipidemia, arteriosclerosis, cardiac disease, diabetes, obesity, digestive organ disease, infectious disease, allergic disease, endocrine disease, dementia or cancer, or complications thereof; and provides a nonhuman transgenic mammal for the unraveling of the mechanisms of said diseases, explorations for the development of therapeutic modalities for the diseases, and the screening of candidate therapeutic drugs.

Abstract: The present invention provides an isolated nucleic acid encoding a mammalian capping enzyme. The present invention further provides an isolated nucleic acid encoding a mammalian (Guanine-7-) methyltransferase enzyme. The present invention also provides an isolated mammalian capping enzyme protein or subunit thereof. In addition the present invention provides an isolated mammalian (Guanine-7-) methyltransferase enzyme protein or portion thereof. The present invention further provides methods for catalyzing formation of RNA 5′-terminal GpppN cap complex and for coupled transcription, translation and formation of RNA 5′-terminal GpppN cap complex and methylated RNA 5′-terminal GpppN cap complex. Kits thereto are also provided.

Abstract: The present invention provides a general method for identifying genes and producing immunological reagents which encode cell surface antigens of human origin. Specifically, this invention provides a method for preparing a hybridoma cell line which produces an antibody which specifically recognizes and binds to a cell surface antigen associated with a neoplastic, human cell. This invention also provides a method for preparing a monoclonal and a polyclonal antibody which specifically recognizes and binds to a cell surface antigen associated with a neoplastic, human cell. Using the antibodies produced, this invention further provides a method for diagnosing in a subject a neoplastic condition, a method for treating a neoplastic condition, and a method for imaging a neoplastic, human cell.

Abstract: The present invention provides, inter alia, a PC12 cell quiescently infected with a neurotropic virus. Preferred quiescently-infected neural cells are those wherein the neurotropic virus is a neurotropic herpes virus. Methods related to these and other materials are also provided.

Abstract: Carboxymethylation of proteins is a highly conserved means of regulation in eukaryotic cells. The protein phosphatase 2A (PP2A) catalytic (C) subunit is reversibly methylated at its carboxy-terminus by specific methylesterase. Carboxymethylation affects PP2A activity and varies during the cell cycle. The present disclosure provides the coding sequence of a methylesterase, herein named Protein Phosphatase Methylesterase-1 (PME-1). PME-1 is highly conserved from yeast to human and contains a motif found in lipases, which motif has a catalytic triad-activated serine as the active site nucleophile. Recombinant PME-1 polypeptide produced in bacteria demethylates PP2A C subunit in vitro and okadaic acid, a known inhibitor of the PP2A methylesterase, inhibited this reaction. PME-1 represents the first mammalian protein phosphatase methylesterase cloned to date.

Abstract: Methods of treating individuals suspected of suffering from diseases, conditions or disorders of the Central Nervous System which comprise implanting stable, homogeneous post-mitotic human neurons into the individual's brain are disclosed. Methods of treating individuals suspected of suffering from injuries, diseases, conditions or disorders characterized by nerve damage which comprise implanting stable, homogeneous post-mitotic human neurons at or near a site of said nerve damage are also disclosed.

Abstract: The DNA encoding the cell surface receptor for thrombin has been cloned and sequenced. The availability of this DNA permits the recombinant production of thrombin receptor which can be produced at cell surfaces and is useful in assay systems both for the detection of thrombin and for the evaluation of candidate thrombin agonists and antagonists. Further, the elucidation of the structure of the thrombin receptor permits the design of agonist and antagonist compounds which are useful diagnostically and therapeutically. The availability of the thrombin receptor also permits production of antibodies specifically immunoreactive with the receptor per se or with specific regions thereof which are also useful diagnostically or therapeutically.

Abstract: This invention provides a CREF-Trans 6 cell line. The invention further provides CREF-Trans 6 cell lines containing DNA from an established cancer cell line or a primary tumor. This invention also provides the use of said cell lines to obtain DNA encoding a cell surface antigen associated with a neoplastic human cell line.

Abstract: The invention features immortalized retina-derived (retinal endothelial or retinal epithelial pigmentary) cell lines capable of being implanted in the retina and of carrying a therapeutic substance to the eye and to the central nervous system. Such lines can also be used as a model for studying blood central nervous system interfaces. These lines are derived from primary retinal cultures selected from the group consisting of primary retinal endothelial cells and primary retinal epithelial cells, comprise a nucleic acid fragment containing at least one immortalizing fragment of a heat-sensitive viral oncogene, which nucleic fragment is optionally associated with at least one selection gene, and have the morphological characteristics and at least the expression characteristics of the surface antigens of corresponding primary cultures.

Abstract: The replication machinery of polio virus is used to express heterologous gene products, such as chloramphenicol acetyl transferase, in mammalian cells. Detectable expression following DNA transfection demonstrated that a polio replicon containing a foreign gene in the P1 region transcribed from an inducible promoter can be exported from the nucleus to the cytoplasm. The proteins in the P2/P3 region of the RNA can be translated and thereby render the RNA capable of replication. A stable cell line harbouring the polio replicon in the genome results in constitutive expression of chloramphenicol acetyl transferase or other heterologous gene product.

Abstract: Mammalian stem cells are obtained and maintained in vitro whose genome has at least one nucleic acid construct encoding an antibiotic resistance gene operatively linked to a promoter specific for mammalian stem cells. The preferential expression of the antibiotic resistance gene in the stem cells results in the preferential survival of the stem cells in the presence of antibiotic.

Abstract: Immortalized cell lines of retinal origin (retinal endothelial and retinal pigmentary epithelial origin) which are capable of being implanted in the retina and of conveying a substance of therapeutic interest into the eye and the central nervous system. Such lines can also serve as a model for studying the blood/central nervous system interfaces.These lines are derived from primary cultures of retinal cells selected from the group comprising the primary retinal endothelial cells and the primary retinal epithelial cells, comprise a nucleic acid fragment containing at least one immortalizing fragment of a heat-sensitive viral oncogene, which nucleic acid fragment may be associated with at least one selection gene, and exhibit the morphological characteristics and at least the surface antigen expression characteristics of the corresponding primary culture cells.

Abstract: Ligands which bind to the eck receptor are disclosed. More particularly, polypeptides which bind specifically to the eck receptor (eck receptor binding proteins or EBPs) and DNA sequences encoding said polypeptides are disclosed. Methods of treatment using eck receptor ligands and soluble eck receptor and disclosed, as are pharmaceutical compositions containing same. A rapid and sensitive method for the detection of receptor binding activity in crude samples is provided.

Abstract: A composition and method for inhibiting the growth and metastasis of breast tumors, including a tumor immunogen derived from breast tumor cells treated with an antisense oligonucleotide complementary to a gene or mRNA for the receptor for insulin-like growth factor type 1.

Abstract: The 5'-flanking region and core regulatory domains that underlie neuronal specific expression of the human .gamma.-aminobutyric acid type A (GABA.sub.A) receptor .beta.1 subunit gene are identified herein. Sequence analysis, mapping of transcriptional initiation sites, and transfection of reporter gene constructs into primary cultures demonstrate that neuronal and region specific activity resides in a TATA-less minimal promoter of 186 bp, comprising an initiator, the major transcriptional start site, a presumptive TFIID binding site, and an enhancer. Enhancer sequence contained within a 26 bp region at the 5'-end of the minimal promoter is essential for activity but not for tissue specificity. Moreover, .beta.1 promoter activity is subject to autologous inhibition, indicating that GABA-induced receptor mRNA downregulation results from an inhibition of gene transcription.

Abstract: The present invention reveals in vitro cultures of region-specific, terminally differentiated, mature neurons derived from cultures of mammalian multipotential CNS stem cells and an in vitro procedure by which the differentiated neurons may be generated. The procedure involves the culturing of multipotential CNS stem cells from a specific region in a chemically defined serum-free culture medium containing a growth factor; replacing the medium with growth factor-free medium; harvesting the stem cells by trypsinization; plating the stem cells at a density of between 100,000 to 250,000 cells per square centimeter; and culturing the stem cells in a glutamic acid-free chemically defined serum-free culture medium.

Abstract: Method for producing a population of mammalian glial cells comprising contacting at least one mammalian neural stem cell with a culture medium containing a neuregulin and detecting the differentiation of stem cell to a population of glial cells.

Abstract: The present invention provides liver parenchymal cells having a clonal growth ability, which possesses at least one of the cell biological properties such as: presence of peroxysome; being positive to hepatocyte-markers; being partially positive to neoplastic hepatocyte-markers or immature hepatocyte-markers; being positive to antibodies against the surface antigens of ovall cells; and being partially positive to bile duct cell-markers. The present invention also provides a method for obtaining such cells and a method for subculturing such cells.With the liver parenchymal cells above, it will be possible to research in detail the process of development and differentiation, the mechanisms of growth and functional expression of hepatic cells, and to open up a new way to clalification of mechanisms or hepatoma and various other diseases and to development of therapeutic method against these disseases.

Abstract: Method for producing a population of mammalian neurons and/or smooth muscle cells comprising contacting at least one mammalian neural stem cell with a culture medium containing one or more growth factors from the TGF-.beta. super family and detecting the differentiation of stem cell to a population of neurons or smooth muscle cells.

Abstract: A method for identifying anti-convulsant compounds is presented. The binding of test compounds with a receptor obtained from rat forebrain is determined. The receptor has a Kd of about 40 nM and a molecular weight of about 130 kD. Two pyran compounds that bind to the receptor are also disclosed.

Abstract: A myeloma cell-line transformed with a vector including a gene coding for a eukariotic polypeptide and a non-immunolglobulin promoter such that expression occurs of the gene coding for the eukariotic polypeptide, directed by the non-immunoglobulin promoter. The promoter may be a viral promoter, such as an SV40 promoter, a Rous sarcoma virus long terminal repeat or a Moloney murine leukemia long terminal repeat, or a non-viral promoter such as the mouse metallothionein promoter. Rat and mouse host myeloma cell-lines such as the rat YB/2/3.0 Ag20 hybridoma, the mouse SP-20 Ag hybridoma and the mouse NSO hybridoma are employed. The production of tissue plasminogen activator (tPA) is exemplified.

Abstract: Leukemia inhibitory factor or LIF as an addition to culture media promotes the development of embryos to the implantation stage. Growth in the presence of LIF increases the percentage of embryos that reach the implantation stage than growth without LIF. LIF is isolatable from several mammalian species such as murine, sheep, pig, cow, horse or donkey.

Abstract: An isolated and stable clone of mesangial cells which exhibit characteristics of diabetic cells when grown in normal glucose medium. The cells demonstrate an increase in glucose transport over normal mesangial cells and that provides a model for simulating diabetes in mesangial cells. Further, a stable, permanent clone of mesangial cells which underexpress GLUT1 mRNA and protein and which can be used therapeutically has been isolated.

Abstract: Method for the preparation and conservation of cellular cultures or parts of cells, whereby the cellular cultures or parts of cells, after having been subjected to a step of starvation (12), undergo a process of conservation (24) without a significant loss of vitality or function and can therefore be supplied in the ready state for biological dosages.

Abstract: Substrates containing a copolymer of sequentially alternating lysine and alanine residues are provided. The use of those substrates for promoting neuron survival and axon growth are also provided.

Abstract: The invention relates to the field of biology and, in particular, to the field of cellular biology. The invention concerns a novel glucose-sensitive cell line designated .beta. cell line (INS-I) expressing glucokinase and the glucose carrier Glut 2 at levels comparable with those of normal .beta. cells but which is, in addition, incapable of IGF-II expression-dependent proliferation because of genetic manipulation. The invention also concerns a method for the production of said novel cell line, its aggregation in the form of a pseudoislet, its immobilization in a biocompatible hydrogel and its hardening by means of a hardening solution. Application in insulin-secreting .beta.-cell transplants.

Abstract: Transformed cell lines containing a reporter gene operatively linked to a genetic control element that is responsive to growth factor-stimulated cell proliferation and/or oncogene-mediated neoplastic transformation are provided. Also provided are methods for using such transformed cell lines to screen for growth factor antagonists and/or antineoplastic agents.

Abstract: A gene activation/inactivation and site-specific integration system has been developed for mammalian cells. The invention system is based on the recombination of transfected sequences by FLP, a recombinase derived from Saccharomyces. In several cell lines, FLP has been shown to rapidly and precisely recombine copies of its specific target sequence. For example, a chromosomally integrated, silent .beta.-galactosidase reporter gene was activated for expression by FLP-mediated removal of intervening sequences to generate clones of marked cells. Alternatively, the reverse reaction can be used to target transfected DNA to specific chromosomal sites. These results demonstrate that FLP can be used, for example, to mosaically activate or inactivate transgenes for a variety of therapeutic purposes, as well as for analysis of vertebriate development.

Abstract: The present invention relates, in general, to a gene transfer system and, in particular, to an adenoviral vector system suitable for use in gene transfer and gene therapy.

Abstract: The present invention relates to two groups of suicide hybrid genes in which genes from one group specifically activate the pyrimidine nucleobase analog 5-fluorocytosine and genes from the other group activate the pyrimidine nucleoside analog azidothymidine to derivatives toxic for mammalian cells.The present invention further relates to methods for the selective killing of transfected tumor cells or immune cells using a single suicide hybrid gene or the combination of two suicide hybrid genes selected for a complementarity in their antimetabolic action. The present invention also relates to eukaryotic vectors comprising two expression suicide gene units, the first permitting the sensitization of tumor cells to 5-fluorocytosine and the second permitting the sensitization of HIV-infected cells to Azidothymidine in a synergistic fashion.

Abstract: Disclosed are (1) nucleic acid and amino acid sequences for a novel morphogenic protein; (2) methods for producing and expressing the protein in a biologically active form; and (3) methods for utilizing the protein to induce tissue morphogenesis in a mammal, including methods for increasing a progenitor cell population in a mammal, methods for stimulating progenitor cells to differentiate and maintain their differentiated phenotype in vivo or in vitro, methods for inducing tissue-specific growth in vivo and methods for the replacement of diseased or damaged tissue in vivo.

Abstract: The invention includes mammalian multipotent neural stem cells and their progeny and methods for the isolation and clonal propagation of such cells. At the clonal level the stem cells are capable of self regeneration and asymmetrical division. Lineage restriction is demonstrated within developing clones which are sensitive to the local environment. The invention also includes such cells which are transfected with foreign nucleic acid, e.g., to produce an immortalized neural stem cell, and immortalized cell lines which are capable of subsequent disimmortalization. The invention further includes transplantation assays which allow for the identification of mammalian multipotent neural stem cells from various tissues and methods for transplanting mammalian neural stem cells and/or neural or glial progenitors into mammals. A novel method for detecting antibodies to neural cell surface markers is disclosed as well as a monoclonal antibody to mouse LNGFR.

Abstract: A preventive or therapeutic agent for Alzheimer's disease which comprises a substance exhibiting an inhibitory action to tau-protein kinase I as an effective component is provided. A pharmaceutical composition comprising said agent and a method of inhibiting neuronal cell death in the brain are also provided.

Abstract: This invention relates to the identification and molecular characterization of NAD:arginine ADP-ribosyltransferases. Sequences from the rabbit skeletal muscle NAD:arginine ADP-ribosyltransferase and the human NAD:arginine ADP-ribosyltransferase are provided herein. Recombinant protein is expressed from a recombinant gene vector containing at least 15 continuous bases of genes encoding these sequences.

Abstract: The present invention provides methods for reducing a pathology in a subject characterized by a deleterious accumulation of TGF-.beta.-induced extracellular matrix in a tissue by introducing a nucleic acid encoding a TGF.beta.-specific inhibitory agent or active fragment thereof into a cell in the subject. In one embodiment, the nucleic acid encoding the TGF-.beta. specific inhibitory agent is introduced into a cell in vivo by injection, for example, in skeletal muscle. In another embodiment, the nucleic acid encoding the TGF-.beta. specific inhibitory agent is transfected into a cell ex vivo to obtain a cell expressing the agent, and the cell is then administered into the subject to be treated. TGF-.beta. specific inhibitory agents of the present invention include, but are not limited to, members of the decoring family of proteoglycans such as decorin, biglycan, fibromodulin and lumican or an antibody specific for TGF-.beta..

Abstract: A novel method and test kit for mutagenesis testing applicable to an organism or a cell.

Abstract: The invention relates to virally-encoded nucleic acid sequences derived from a region 5' to the transcriptional start site of the virus which contain at least one direct repeat of a fragment of a hormone-responsive element. In a preferred embodiment, the virus is murine mammary tumor virus, and the hormone-responsive element is a glucocorticoid responsive element.

Abstract: Oligomers which inhibit expression of a collagen gene are described. It is believed that each oligomer, when introduced into a cell, forms a transcription-inhibiting complex composed of the oligomer and the collagen-gene promoter region. The oligomer, preferably a phosphorothioate deoxyoligonucleotide, preferably binds in the antiparallel orientation to the polypurine strand of a polypurine-polypyrimidine region of the promoter region of a mammalian .alpha.1(I) collagen gene.

Abstract: The present invention is directed to isolated nucleic acid molecules encoding gamma glutamyl hydrolase (GH). Expression vectors and host cells comprising the nucleic acid molecules are also provided, as well as methods for increasing or decreasing the expression of GH in host cells. The invention further provides a method of screening a substance for the ability of the substance to modify GH function, and a method for isolating other GH molecules. DNA oligomers and antibodies specific for GH are provided, each of which can be used to detect GH in a sample. Methods for decreasing deleterious side effects of antifolate treatment, increasing the levels of GH in cells of a patient, increasing the effectiveness of antifolate treatment, and monitoring progression of a tumor are further provided.