It is estimated that approximately 30% of men have reduced fertility and 2% are totally infertile. Despite these large numbers relatively little is know about the molecular bases of male infertility. On the flip side of male infertility is the need for male contraception. Currently there are no reversible, convenient male contraceptives available. In order to develop male contraceptives and acquire a greater understanding of male fertility there is a need to develop animal models to study the molecular basis and pathways that regulate and control male fertility. Vanderbilt researchers have developed a model mouse system to study male fertility. There research focuses on the epididymus, which is the area that spermatozoa acquire the ability to move and fertilize. For this region to be functional tissue and cell specific gene regulation must occur. These investigators have discovered one such gene regulated within this area, mEP17. These researchers can fuse either mouse or human EP17 or just the regulatory regions of either EP17 to reporter genes and the resulting fusion can be used to screen for substances that regulate this gene and affect male fertility. This system becomes a powerful tool to identify drugs which affect this gene and be potential male contraceptives. In addition polypeptides generated to this gene may be used as vaccines for male contraceptives.
This invention is directed to cloning and characterization of BVES (blood vessel/epicardial substance), a cDNA expressed in developing and adult heart and skeletal muscle cells in chick, mouse and human. Also provided are applications of BVES as a marker for cardiovascular or skeletal muscle diseases.
A method of testing a candidate composition for PAI-1 inhibition activity is disclosed. The method includes the steps of obtaining a transgenic non-human warm blooded vertebrate animal having incorporated into its genome a PAI-1 gene encoding a biologically active PAI-1 polypeptide, the PAI-1 gene being present in the genome in a copy number effective to confer over-expression in the transgenic non-human animal of the PAI-1 polypeptide; administering the composition to the transgenic non-human animal; and observing the transgenic non-human animal for determination of a change in the transgenic non-human animal indicative of inhibition of the activity of PAI-1. A transgenic non-human animal useful in such a method is also disclosed, as is a PAI-1 transgene construct encoding a biologically active PAI-1 polypeptide useful for preparing the transgenic non-human animal.
An isolated nucleic acid encoding the Helicobacter pylori vacuolating toxin, consisting of the nucleotides 101 through 3964 of the nucleotide sequence defined in the Sequence Listing as SEQ ID NO:1 is provided. An isolated nucleic acid from Helicobacter pylori comprising the nucleotide sequence defined in the Sequence Listing as SEQ ID NO:3 is provided. Isolated nucleic acids that selectively hybridize with the nucleic acids of the invention are provided. Also provided is a genetically altered mutant strain of H. pylori that does not express a functional vacuolating toxin. Purified proteins encoded by the nucleic acids of the invention are provided. A composition comprising an immunogenic amount of a protein or mutant strain of the invention in a pharmaceutically acceptable carrier is provided. A method of immunizing a subject against infection by H. pylori, comprising administering to the subject an immunogenic composition of the invention is provided.
An apparatus comprising one or more piezoelectric mass sensors for use in diagnostic and analytic processes, in particular for immunochemical detection of diagnostically relevant analytes in real time. Each piezoelectric mass sensor comprises a piezoelectric crystal with a receptor surface which has immobilized thereon a lawn of recombinant antibodies comprising single V.sub.H chain or single-chain Fv (scFv) polypeptides specific for a particular antigen. Binding of antigen to the recombinant antibodies results in a change in mass on the receptor surface which is detected as a change in resonant frequency. In a preferred embodiment, the receptor layer is a precious metal such as gold which facilitates self-assembly of the recombinant antibodies into a lawn on the receptor surface via a cysteine residue at the carboxy terminus of the attachment polypeptide.
The invention provides the dapE gene of Helicobacter pylori and H. pylori dapE.sup.- mutants and to methods of using the mutants to express foreign genes and immunize against foreign agents. The dapE gene can consist of the nucleotide sequence defined in SEQ ID NO:3. Nucleic acids of the gidA gene and ORF 2 of H. pylori are provided. Examples of these nucleic acids can be found in SEQ ID NO:1 and SEQ ID NO:5, respectively. Having provided these nucleic acids, hybridizing nucleic acids in accord with the description of hybridizing nucleic acids of dapE are also provided.
The present invention provides methods for screening an EphB receptor or an EphB receptor-binding ligand for the ability to promote a selected biological activity when in multimeric form. The invention also provides methods for initiating, promoting, directing, or inhibiting biological activities that involve EphB receptors and/or EphB receptor-binding ligands. The invention further provides compositions that can be used in the foregoing methods.
The invention provides an isolated nucleic acid encoding the rat P450 2C11 arachidonic acid epoxygenase, or a human homologue thereof, having a mutation associated with salt induced hypertension. Also provided is an isolated cell line expressing the epoxygenase encoded by the mutated nucleic acid, and a non-human transgenic animal having a germ line insertion of the mutated nucleic acid. Also provided is a method of screening a compound for efficacy in treating salt induced hypertension comprising administering the compound to such a non-human transgenic animal, and detecting an improvement in the animal's hypertension. The invention also provides a method of screening a human subject for a genetic predisposition to salt induced hypertension comprising detecting a mutation in a human homologue of a rat P450 2C11 arachidonic acid epoxygenase gene which affects normal epoxygenase activity. Also provided is a method of treating salt induced hypertension in a human subject associated with a genetic mutation in a human homologue of the rat P450 2C11 arachidonic acid epoxygenase gene, comprising administering to the subject a functional metabolite, or analogue thereof, produced by the human homologue of the rat P450 2C11 arachidonic acid epoxygenase. Also provided is an isolated mutated rat P450 2C11 arachidonic acid epoxygenase, or a human homologue thereof, having a mutation associated with salt induced hypertension.
A cagB gene of H. pylori is provided. This nucleic acid can be the nucleic acid consisting of nucleotides 193 through 1158 in the sequence set forth as SEQ ID NO:1, which is an example of a native coding sequence for CagB. This nucleic acid can also be in a vector suitable for expressing a polypeptide encoded by the nucleic acid. A cagC gene of H. pylori is provided. This nucleic acid can be the isolated nucleic acid consisting of nucleotides 1170 through 3830 in the sequence set forth as SEQ ID NO:3, which is an example of a native coding sequence for CagC. This nucleic acid can also be in a vector suitable for expressing a polypeptide encoded by the nucleic acid. Isolated nucleic acids that specifically hybridize with cagB and cagC are provided. CagB and CagC are associated with peptic ulceration and other clinical syndromes in humans infected with strains of H. pylori that express it.
The present invention provides for the use of Tripterygium wilfordii Hook F extracts and purified components thereof in the treatment of inflammation or an immune disorder with concomitant lack of steroidal effect. Extracts of this plant (T2) bound to the glucocorticoid receptor and competitively inhibited glucocorticoid mediated cellular processes, such as dexamethasone binding to the glucocorticoid receptor, glucocorticoid mediated activation of target genes, dexamethasone dependent cellular growth, with concomitant inhibition of cyclooxygenase-2 induction and inflammatory processes such as the production of prostaglandin E.sub.2. The T2 extract components triptolide and tripdiolide were effective inhibitors. The particular advantage provided by the methods herein is the treatment or prevention of inflammation and the concomitant lack of steroidal agonist effects and NSAID side effects. Conditions treatable by the present methods include inflammation and immune disorders including autoimmune disease.
Two rabbit anti-leukocyte-associated immunoglobin-1 (LAIR-1) polyclonal antibodies have been developed. Both intracellular (epitope 170-269) and extracellular (epitope 1-141) antibodies are available which can be used for immunoprecipitation and immunoblotting, respectively.