Abstract: The present invention relates to novel mammalian amino acid transporter proteins and the genes that encode such proteins. The invention is directed toward the isolation, characterization and pharmacological use of the human amino acid transporter proteins EAAT1, EAAT2, EAAT3 and ASCT1. The invention specifically provides isolated complementary DNA copies of mRNA corresponding to each of these transporter genes. Also provided are recombinant expression constructs capable of expressing each of the amino acid transporter genes of the invention in cultures of transformed prokaryotic and eukaryotic cells, as well as such cultures of transformed cells that synthesize the human amino acid transporter proteins encoded therein. The invention also provides methods for screening in vitro compounds having transport-modulating properties using preparations of transporter proteins from such cultures of cells transformed with recombinant expression constructs.

Abstract: The present invention provides methods and reagents for assessing lipoprotein metabolism. The methods provided are applicable for use on multiple samples in a clinical laboratory, thus obviating the need for sophisticated instrumentation, such as flow cytometry. Selected cell types in a test sample are uniformly labelled with a detectable reporter substance, so that they may later be enumerated. Pre-determined lipoprotein receptors associated with the cells of interest are labelled with a receptor-selective marker, thereby to determine the number of lipoprotein receptors per cell in the test sample. Selected cells of interest are conveniently separated from the test sample by binding thereto a specific binding substance attached to a solid support. The specific binding substance binds specifically with a characteristic determinant of the cell subset of interest. The remainder of the test sample may be washed away or discarded.

Abstract: The present invention is directed to screening for an agent that inhibits mononuclear phagocyte-plaque component complex formation (hereinafter "complex formation"). The methods include the steps of contacting a mononuclear phagocyte with a plaque component to stimulate complex formation and adding an agent suspected of inhibiting complex formation, measuring complex formation, and comparing complex formation to a measured control, wherein the reduction of complex formation compared to the control results in detection of an agent that inhibits complex formation. The mononuclear phagocytes may be from mammalian brain. The plaque component may be coupled to a solid support.

Abstract: A method for sequential chemiluminescent detection of two differently labeled analytes on a single blot is described. In the method, a uniquely labeled DNA is detected with a horseradish peroxidase (HRP) substrate followed by the detection of another uniquely labeled DNA with a second different enzyme substrate which also inhibits the chemiluminescence generated by HRP. The sequential detection method described herein eliminates the need to strip and reprobe Southern, Northern and Western blots. Potential applications of this method include forensic DNA fingerprinting where more than one probe is used for probing a Southern blot, multiplex DNA sequencing of more than one template, detection of gene rearrangements, mutations and gene linkage.

Abstract: Methods for simultaneously selecting binding site sequences for multiple DNA-binding proteins are provided. A source of DNA-binding proteins is mixed with oligonucleotide duplexes containing a randomized internal sequence. Bound oligonucleotides are isolated, amplified and analyzed, such as by DNA sequence analysis. The oligonucleotide duplexes are also used to identify and isolate DNA-binding proteins.

Abstract: The present invention relates to screening or testing for insulin like growth factors, insulin like growth factor binding proteins and/or acid labile subunit by the use of a solid support. Blood is collected onto a solid support, such as paper, and subsequently the analytes of interest are extracted for testing.

Abstract: In accordance with the present invention, it has been discovered that CREB binding protein (CBP) cooperates with upstream activators involved in the activation of transcription of such signal dependent transcription factors as c-Jun (responsive to phorbol ester), serum response factor, and the like. It has also been discovered that CBP can be employed in an assay to identify compounds which disrupt the ability of such signal dependent transcription factors to activate transcription. In another aspect, it has been discovered that CBP can be employed in an assay to identify new signal dependent transcription factors. In yet another aspect of the present invention, it has been discovered that CBP can be employed in an assay to identify novel co-factor protein(s) which mediate the interaction between signal dependent transcription factors and inducer molecules involved in the activation of transcription.

Abstract: Conjugated compounds which comprise an ST receptor binding moiety and a radiostable active moiety are disclosed. Pharmaceutical compositions comprising conjugated compound which comprises an ST receptor binding moiety and a radiostable active moiety or an ST receptor binding moiety and a radioactive active moiety are disclosed. Methods of treating an individual suspected of suffering from metastasized colorectal cancer are disclosed. Methods of radioimaging metastasized colorectal cancer cells are disclosed. In vitro methods, kits and reagents are disclosed for determining whether or not an individual has metastasized colorectal cancer cells, for determining whether tumor cells are colorectal in origin and for analyzing tissue samples from the colon tissue to evaluate the extent of metastasis of colorectal tumor cells.

Abstract: This invention provides non-naturally occurring and isolated naturally occurring nucleic acid molecules which encode proteins designated Yama. This invention also provides the polypeptides and proteins encoded by these nucleic acids as well as the purified native polypeptides or proteins. Also provided by this invention is a non-naturally occurring nucleic acid molecule encoding mutant CrmA protein and a dominant inhibitory Yama. Vectors and host cells containing these nucleic acid molecules are further provided. Methods of modulating a cellular function regulated by the Fas receptor pathway in a cell is provided herein. In one aspect, this method comprises introducing into the cell a nucleic acid molecule coding for a gene product having CrmA biological activity such as dominant inhibitory Yama or alternatively, the CrmA gene product. Yama nucleic acid molecules and proteins also can be introduced into the cell to modulate the cellular function regulated by the Fas receptor.

Abstract: The invention is directed to inter alia two related but self-sufficient improvements in conventional display methods. The first improvement provides methods of enriching conventional display libraries for members displaying more than one copy of a polypeptide prior to affinity screening of such libraries with a target of interest. These methods can achieve diverse populations in which the vast majority of members retaining full-length coding sequences encode polypeptides having specific affinity for the target. In a second aspect, the invention provides methods of subcloning nucleic acids encoding displayed polypeptides of enriched libraries from a display vector to an expression vector without the need for clonal isolation of individual members. These methods result in polyclonal libraries of antibodies and other polypeptides for use, e.g., as diagnostic or therapeutic reagents.

Abstract: A method for the detection of a compound which modulates binding of a ligand to a biological receptor, which method comprises contacting the ligand, biological receptor and test compound at a locus on a solid phase matrix, the matrix allowing movement of fluids therein by capillary action, under conditions which permit binding of the ligand to the biological receptor and partition of any unbound ligand on the solid phase matrix, and detecting any modulation of binding by the test compound by reference to any such partition.

Abstract: Disclosed is a method of testing a substance for the ability to affect the formation or oligomerization of a complex comprising members of a specific binding pair, a first member of the binding pair being present on the surface of a lipid enveloped particle comprising a transferrable label, said first member of the binding pair being capable of binding to a second member of the specific binding pair present on the surface of a cell, the lipid envelope of the particle being capable of fusing with the membrane of the cell so as to transfer the label to the cell, said transfer being inhibited by formation or oligomerization of a complex between the first and second members of the binding pair, wherein the method comprises reacting the particle and the cell, in the presence of the substance under test, (under conditions which would allow for binding of the first and second members of the specific binding pair in the absence of the substance under test), and detecting the label transferred, if any; together with a

Abstract: Disclosed is a method for screening for inhibitors of Hepatitis B Virus pX activity. The method involves contacting a test compound with (I) the pX protein of HBV, (ii) a transcription factor comprising the bZIP domain, or fragments that comprise a minimal a bZIP domain, and (iii) an oligoduplex comprising a target DNA sequence of the transcription factor to form a test mixture. After incubating the test mixture under appropriate conditions and for a sufficient time to allow pX-mediated dimerization and DNA binding of the transcription factor to occur, the level of DNA binding of the transcription factor in each test mixture is determined. A test compound is considered to be any compound that causes a decrease in the level of DNA binding in the test mixture relative to the level of DNA binding in control mixtures.

Abstract: The present process for producing a high .alpha.-monoglucosyl hesperidin content product is characterized in that it comprises the steps of contacting glucoamylase and .alpha.-L-rhamnosidase simultaneously or randomly with a solution containing .alpha.-glucosyl hesperidin and hesperidin to obtain a mixture; crystallizing and separating .alpha.-monoglucosyl hesperidin in and from the mixture; and collecting the resulting .alpha.-monoglucsyl hesperidin. From solutions containing .alpha.-glucosyl hesperidin and hesperidin, the present invention facilitates the production of a high .alpha.-monoglucosyl hesperidin content product which does not substantially contain hesperidin, .beta.-monoglucosyl hesperetin, and hesperetin, and has an extremely-superior water-solubility.

Abstract: A novel human serine protein kinase, human p21-protein activated serine kinase p65 protein, referred to as hPAK65, and methods for its preparation and use are provided. Nucleic acids encoding hPAK65 and methods for their use in preparing hPAK65 as well as in preparing and identifying hPAK65 analogs are provided. Methods provided for the use of hPAK65 protein and its protein fragments, such as those that retain at least one hPAK65 activity, that include screening libraries of agents for candidates that modulate hPAK65 activity. Methods are provided to identify agents that modulate the interaction of hPAK65 with rho-like p21 GTPases, particularly rac1 and CDC42Hs binding to hPAK65 and subsequent activation of hPAK65 serine protein kinase activity, that modulate hPAK65 serine protein kinase activity, and that modulate hPAK65 effect on p21 protein GTPase activity.

Abstract: Disclosed are methods and compositions for identifing agents which modulate the interaction of Robo and a Robo ligand and for modulating the interaction of Robo and a Robo ligand. The methods for identifying Robo:ligand modulators find particular application in commercial drug screens. These methods generally comprise (1) combining a Robo polypeptide, a Slit polypeptide and a candidate agent under conditions whereby, but for the presence of the agent, the Robo and Slit polypeptides engage in a first interaction, and (2) determining a second interaction of the Robo and Slit polypeptides in the presence of the agent, wherein a difference between the first and second interactions indicates that the agent modulates the interaction of the Robo and Slit polypeptides.

Abstract: The invention provides a monospecific antibody that is specifically reactive with enzymatically mediated degradation products of fibrin(ogen) (i.e., fibrin, fibrinogen, and related substances). The monospecific antibody of the invention is specifically reactive with an epitope defined by an amino acid sequence SEQ ID NO:1. The invention further provides compositions containing a monospecific antibody, optionally detectably labeled, for the performance of fibrinolytic or thrombolytic analyses. The invention further provides continuous cell lines (hybridomas) that produce monospecific antibodies as described.

Abstract: The present invention derives from the discovery that CDC25 phosphatases and Raf proteins are able to physically interact to form protein-protein complexes, with the Raf protein mediating the activation of CDC25 phosphatases. The present invention provides both cell-free and cellular assays for detecting agents which modulate the ras-dependent activation of CDC25, as for example, by affecting the binding of a CDC25 protein with Raf, or Raf-associated complexes. Also disclosed is a method for transforming/immortalizing cells, particularly primary cell cultures.

Abstract: A non-competitive method for the determination of analytes. Initially the analyte is bound to a specific binding partner, after which the unoccupied binding sites of the binding partner are inactivated. The bound analyte is then dissociated from the binding partner and replaced by a labeled marker, after which the bound labeled marker is determined. The signal from the bound labeled marker is directly proportional to the initial amount of analyte in the sample, which makes the present method more favorable than the competitive assays.

Abstract: There are provided: an immunological reagent for diagnosis of or the evaluation of the effect of a medicine for dialysis-related amyloidosis, which includes an antibody which reacts specifically with a carboxymethylated .alpha.-amino acid, a carboxymethylated protein, or a carboxymethylated peptide; or, an immunological reagent for diagnosis of or the evaluation of the effect of a medicine for diabetes mellitus and diabetes mellitus complications, which includes an antibody which reacts specifically with an N.sup..alpha. -carboxymethylated .alpha.-amino acid, an N.sup..alpha. -carboxymethylated protein, or an N.sup..alpha. -carboxymethylated peptide. Methods using the immunological reagents are provided: for determination of carboxymethylated hemoglobin as a marker for dialysis-related amyloidosis; or, for determination of N.sup..alpha. -carboxymethylated hemoglobin as a marker for diabetes mellitus or diabetes mellitus complications.

Abstract: The present invention relates to the finding that cyclin D1 interacts with estrogen receptor to provide activation of estrogen responsive genes. The present invention provides in vitro and in vivo assays to measure the interaction. The in vivo assays may be conducted in cells which grow in response to estrogen, particularly breast tumour cells.

Abstract: An immunoassay method in which blood can be measured even without pretreatment by a centrifuge etc. In the present invention, antibodies or antigens in a sample are subjected to agglutination reaction with insoluble carriers onto which antigens or antibodies specifically reacting with the antibodies or antigens in the sample have been imrobilized and the resulting agglutination mixture is determined for the change in its absorbance or in its scattered light by irradiation with light, wherein said sample is whole blood and the whole blood is forcibly lysed.

Abstract: The invention provides methods and compositions relating to CPF proteins which regulate transcriptional activation, and related nucleic acids. The polypeptides may be produced recombinantly from transformed host cells from the disclosed CPF encoding nucleic acids or purified from human cells. The invention provides isolated CPF hybridization probes and primers capable of specifically hybridizing with the disclosed CPF genes, CPF-specific binding agents such as specific antibodies, and methods of making and using the subject compositions in diagnosis, therapy and in the biopharmaceutical industry.

Abstract: Combinations, called matrices with memories, of matrix materials with remotely addressable or remotely programmable recording devices that contain at least one data storage unit are provided. The matrix materials are those that are used in as supports in solid phase chemical and biochemical syntheses, immunoassays and hybridization reactions. The matrix materials may additionally include fluophors or other luminescent moieties to produce luminescing matrices with memories. The data storage units are non-volatile antifuse memories or volatile memories, such as EEPROMS, DRAMS or flash memory. By virtue of this combination, molecules and biological particles, such as phage and viral particles and cells, that are in proximity or in physical contact with the matrix combination can be labeled by programming the memory with identifying information and can be identified by retrieving the stored information. Combinations of matrix materials, memories, and linked molecules and biological materials are also provided.

Abstract: Netrin proteins, nucleic acids which encode netrin proteins and hybridization reagents, probes and primers capable of hybridizing with netrin genes and methods for screening chemical libraries for lead compounds for pharmacological agents are provided.

Abstract: The invention relates to novel methods and compositions for the detection of analytes using the nuclear reorganization energy, .lambda., of an electron transfer process.

Abstract: The invention relates to novel methods and compositions for the detection of analytes using the nuclear reorganization energy, .lambda., of an electron transfer process.

Abstract: A trans-activation assay for leptin is described. Leptin response elements are located proximal to a promoter, and the promoter region is operatively linked to a reporter gene. When leptin binds with the receptor, the reporter gene is transcribed.

Abstract: sigF is a gene that controls M. tuberculosis latency. A diagnostic test for latent tuberculosis involves detecting M. tuberculosis sigF in clinical specimens. Two genes orfX and orfY regulate sigF expression and sigF activity. M. tuberculosis sigF, orfX, and orfY are used in screening methods for potential therapeutic agents which regulate the growth of M. tuberculosis.

Abstract: Disclosed are compositions and methods for diagnosing autoimmune insulin dependent diabetes mellitus. In general, diagnostic methods of the invention include testing for the presence of an autoimmune immunoglobulin in a suspected patient's serum wherein the immunoglobulin is identified by its ability to interfere with the glucose transporting activity of a pancreatic islet cell glucose transporter. The inventors have discovered that the presence of such an autoimmune antibody in patient's serum is diagnostic of autoimmune insulin dependant diabetes mellitus. In particular aspects the diagnostic assay of the invention involves the incubation of isolated and dispersed islet cells, such as rat islet cells, in the presence of immunoglobulin obtained from the patient. Following such an incubation, the islet cells are tested for their ability to uptake glucose. A diagnosis is made through a determination that immunoglobulins in the serum of the patient specifically inhibit the uptake of glucose by the islet cells.

Abstract: Chemiluminescent electron-rich aryl-substituted 1,2-dioxetane compounds are disclosed in which the aryl group is poly-substituted with suitable electron-donating groups such that the light-emitting pattern of the molecule results in a very high luminescent count, thus providing for a sensitive and precise assay for haptens, analytes, polynucleotides and the like. These substituted aryl-containing 1,2-dioxetane compounds can be used as direct labels in an immunoassay or when derivatized with an appropriate leaving group, can be used as a substrate for a enzyme immunoassay. The unusual chemiluminescence of the compounds allows the timing of the luminescent reaction to be exactly controlled.

Abstract: The invention provides methods and compositions relating to novel bioactive derivatives of indole-3-carbinol (I3C), including pharmaceuticals comprising a pharmaceutically acceptable excipient and a compound of the general formula: ##STR1## which inhibits tumor cell growth. Methods of inhibiting targeted cell growth include contacting a target cell with a disclosed compound under conditions whereby the growth the target cell is inhibited, and methods for evaluating the growth inhibitory activity of the compounds include contacting a cell with an effective amount of the compound and measuring the CDK6 expression in the cell, wherein a reduction in CDK6 expression correlates with the growth inhibitory activity of the compound.

Abstract: This invention relates to novel mammalian excitatory amino acid transporter proteins and genes encoding such proteins. The invention is directed towards the isolation, characterization and use of human excitatory amino acid transporter proteins for pharmacological screening of analogues, agonists, antagonists, inhibitors, modulators and facilitators of excitatory amino acid transport in a variety of tissues, particularly neuronal tissues. This invention provides isolated nucleic acid encoding a novel excitatory amino acid transporter subtype that is specifically expressed in retina. Also provided are recombinant expression constructs capable of expressing this novel transporter in transformed prokaryotic and eukaryotic cells, and also provides such transformed cell cultures producing the novel human transporter. Purified transporter protein and membranes comprising the transporter protein are also provided.

Abstract: The present invention is directed to a process for improving daunorubicin and doxorubicin production by means of a recombinant microorganism in which a gene of daunorubicin metabolism involved in the glycosylation of daunorubicin to acid-sensitive, baumycin-like compounds is inactivated.

Abstract: A test device, system and method, the device composed of an elongated, toothbrush-shaped, transparent, plastic housing and a lateral-flow test strip for the detection of an analyte, such as a beta-lactam in milk, in the housing, the housing having an expansion cavity to receive expanded, liquid-contacted, absorbing material in the test strip, and to control lateral flow rate and times in the test strip.

Abstract: A phagemid has been constructed that expresses an antibody fused to coliphage pIII protein. The phagemid is suitable for selecting specific antibodies from large gene libraries with small quantities of antigen. The antibody-pIII gene can be strongly repressed, so that it allows antibody libraries to be amplified without the danger of deletion mutants predominating. After induction, large quantities of the fusion protein may be expressed.

Abstract: The present invention is directed to a method of identifying plant proteins that function as or similar to G protein coupled receptors, or as plant G protein subunits utilizing a bioassay system that incorporates such plant proteins. Further aspects of the present invention provides expression vectors and yeast cells transformed therewith encoding the plant proteins whose identity it is desired to determine, and methods utilizing same.

Abstract: The present invention provides novel methods for producing doxorubicin using daunomycin as a substrate. One method employs a genetically engineered host microorganism which is transformed with a vector, preferably a plasmid, which contains the doxA gene. Preferably, the doxA gene, also referred to herein as "doxA", is cloned into a plasmid which is then introduced into the host microorganism, preferably a bacterial host, more preferably Streptomyces, to provide a transformed host microorganism. The doxA gene, when present on a plasmid, confers on the transformed host the ability to convert daunomycin and 13-dihydrodaunomycin, to doxorubicin. The doxA gene encodes a P450-like enzyme which catalyzes the hydroxylation of daunomycin and 13-dihydrodaunomycin at C-14 to form doxorubicin; such enzyme is designated "daunomycin C-14 hydroxylase". Thus, the expression of doxA in the transformed host using a plasmid which contains doxA enables the transformed host to convert daunomycin to doxorubicin.

Abstract: A method for diagnosing an HIV-2 (LAV-II) infection and a kit containing reagents for the same is disclosed. These reagents include cDNA probes which are capable of hybridizing to at least a portion of the genome of HIV-2. In one embodiment, the DNA probes are capable of hybridizing to the entire genome of HIV-2. These reagents also include polypeptides encoded by some of these DNA sequences.

Abstract: The present invention relates to the discovery, identification and characterization of nucleotides that encode Ob receptor (ObR), a receptor protein that participates in mammalian body weight regulation. The invention encompasses obR nucleotides, host cell expression systems, ObR proteins, fusion proteins, polypeptides and peptides, antibodies to the receptor, transgenic animals that express an obR transgene, or recombinant knock-out animals that do not express the ObR, antagonists and agonists of the receptor, and other compounds that modulate obR gene expression or ObR activity that can be used for diagnosis, drug screening, clinical trial monitoring, and/or the treatment of body weight disorders, including but not limited to obesity, cachexia and anorexia.

Abstract: Nucleic acids are compacted, substantially without aggregation, to facilitate their uptake by target cells of an organism to which the compacted material is administered. The nucleic acids may achieve a clinical effect as a result of gene expression, hybridization to endogenous nucleic acids whose expression is undesired, or site-specific integration so that a target gene is replaced, modified or deleted. The targeting may be enhanced by means of a target cell-binding moiety. The nucleic acid is preferably compacted to a condensed state.

Abstract: The present invention provides novel assays for determining the ability of a test compound to inhibit intercellular adhesion mediated by a selectin receptor, such as the LHR. The assay involves coating a surface of a solid substrate with an antibody which specifically binds a selectin ligand. Preferred are antibodies against GlyCAM-1 which is a natural biological ligand for L-Selectin and also binds to P-Selectin and E-Selectin. A compound such as GlyCAM-1 is then bound to the antibodies. The test compound mixture putatively containing a selectin ligand is then brought into contact with a chimeric molecule. The test mixture might include any compound which might block or hinder binding between a selectin receptor and the ligand bound to the antibody.

Abstract: The present invention relates to compound 0624 having excellent antitumor activity and antimicrobial activity, which is represented by the general formula (1), and a pharmaceutically acceptable salt thereof. ##STR1## (wherein R represents H or COCH.sub.3.

Abstract: Fluorescent dyes possess reactive linkers for conjugating to nucleic acids, carbohydrates and peptides. The conjugates fluoresce in the visible and UV spectrum and have an excellant solvochromatic response as compared to other fluorescence or chromatic labels. The conjugates are stable but also have medium sensitive. The fluorescent dyes have little triplet state formation and are not photoreactive, making them an excellent substance for biological investigations. Uses for the dyes include protein labelling, DNA labelling, single molecule spectroscopy and fluorescence. A synthesis of the dyes is disclosed. Methods of use include the detection of carbohydrate-protein interactions.

Abstract: The present invention concerns a process for improving doxorubicin production by means of a recombinant Streptomyces peucetius strain bearing a mutation in the gene dnrU coding for a protein involved in the metabolism of daunorubicin.

Abstract: The present invention relates to a method for assessing the ability of a compound to modulate the formation of a complex between a potassium channel and a protein tyrosine kinase. The method comprises the steps of (1) contacting a first polypeptide comprising the proline-rich binding region of the potassium channel, a second protein comprising the SH3 binding domain of the protein tyrosine kinase and the compound to be assessed; and (2) measuring the extent of complex formation between the first polypeptide and the second polypeptide.

Abstract: A receptor for an insulin-like polypeptide is purified from yeast membranes. This "insulin receptor-like protein" has a structure analogous to that of the mammalian insulin receptor. The insulin receptor-like protein of Saccharomyces cerevisiae is a heterotetrameric glycoprotein. The protein has two polypeptide types, a first polypeptide which binds insulin and has an apparent molecular weight of 135,000 to 145,000 daltons and a second polypeptide which has an apparent molecular weight of 90,000 to 95,000 daltons and is phosphorylated on tyrosine in response to binding of insulin by said first polypeptide. The first and second polypeptides are joined by disulfide linkage. The protein requires a divalent metal ion for tyrosine autophosphorylation in response to binding of insulin. The yeast insulin receptor-like protein binds human insulin with a dissociation constant of K.sub.d =8.times.10.sup.-10 M and binds human insulin-like growth factor 1 with a K.sub.d =4.times.10.sup.-10 M.

Abstract: The invention provides a method for determining platelet function in a mammal comprising providing platelets from the mammal, contacting the platelets in suspension with at least one immobilised ECM protein or an effective fragment or analog thereof while applying to the platelets an effective mechanical stimulus for an effective period of time and determining the platelet activation produced.The invention provides a method for monitoring the efficacy of pharmacological agents affecting platelet function in vivo and in vitro.

Abstract: Receptors for the zonula occludens toxin of Vibrio cholera, as well as methods involving the use of the same are disclosed.

Abstract: Enzymatic hydrolysis and glycosidation methods for the preparation of pradimicin compounds, especially the compound BMY-28960: ##STR1## and salts thereof.