Abstract: Endothelin-1 (ET-1), a 21-residue vasoactive peptide, is produced in vascular endothelial cells from the 38-residue inactive intermediate, big endothelin-1 via a specific cleavage at Trp21-Val22. The protease that catalyzes the conversion, endothelin converting enzyme (ECE), constitutes a potential regulatory site for the production of the active peptide. Disclosed herein is the identification of ECE-1, a novel membrane-bound neutral Zn.sup.2+ metalloprotease that is expressed abundantly in endothelial cells in vivo, and structurally related to neutral endopeptidase 24.11 and Kell blood group protein. When transfected into cultured cells that normally secrete only big ET-1, the ECE-1 cDNA conferred the ability to secrete mature ET-1. In transfected cells, ECE-1 processes endogenously synthesized big ET-1 as well as exogenously supplied big ET-1, which interacts with ECE-1 on the cell surface.

Abstract: A method of diagnosing prostate metastasis is provided by the present invention whereby RNA from a patient's blood is isolated and amplified using a pair of primers which are complementary to regions of the prostate specific antigen gene. The presence or absence of amplified RNA is detected and the presence of amplified RNA is indicative micrometastasis of prostate cancer.

Abstract: A method for the detection of a target polynucleotide sequence wherein the target polynucleotide sequence is used as a template to direct hybridization of first and second polynucleotide probes to contiguous portions of the target. The hybridized probes are then ligated, preferably by covalent linkage of photoreactive functional groups present on each of the probes, to produce a ligated reaction product. The ligated reaction product is then multiplied with a polymerase such as Q-beta replicase to produce an enzyme reaction product which is then detected. The presence of the enzyme reaction product is indicative of the presence of the target polynucleotide sequence.

Abstract: The present invention involves a method of amplifying RNA by producing complementary DNA (cDNA) by reverse transcription of RNA, and amplification of the cDNA sequences. The analysis of the amplified material facilitates the detection of pathogens and disease states associated with the presence of particular nucleic acid sequences, so the present invention is important in medical diagnostic procedures. A method of producing cDNA of predetermined length is also disclosed.

Abstract: The present invention relates to the acid sphingomyelinase gene and to methods of diagnosing Niemann-Pick disease. It is based, at least in part, on the cloning and expression of the full-length cDNA encoding acid sphingomyelinase, the cloning and characterization of the genomic structure of the acid sphingomyelinase gene, and on the discovery of frequent mutations in the acid sphingomyelinase gene of Ashkenazi Jewish Niemann-Pick disease patients.

Abstract: A process for detecting a target nucleic acid, which includes the steps of hybridizing a target single-stranded nucleic acid in a sample solution to a labeled single-stranded nucleic acid probe to form a reaction mixture containing a hybrid, subjecting the reaction mixture to capillary electrophoresis to separate the hybrid from the free labeled nucleic acid probe, and detecting the hybrid utilizing the label moiety of the nucleic acid probe constituting the hybrid.

Abstract: Oligonucleotides for and a method of diagnosing prostate micrometastasis are provided by the present invention whereby nucleic acids from a tissue sample from a patient are isolated, nucleic acids from the tissue sample specific for prostate cancer are amplified, or a signal generated by hybridization of a probe specific to a prostate cancer specific nucleic acid is amplified; and detection of amplified nucleic acids is indicative of micrometastasis of prostate cancer.

Abstract: The invention provides passivated organic polymer supports, processes for their preparation and processes for their use in oligonucleotide synthesis that allow for highly efficient solid phase synthesis of oligonucleotides.

Abstract: A method for comparing amounts or levels mRNAs employs a polymerase amplification method using at least two oligodeoxynucleotide primers. In one approach, the first primer contains sequence capable of hybridizing to a site immediately upstream of the first A ribonucleotide of the mRNA's polyA tail and the second primer contains arbitrary sequence. In another approach, the first primer contains sequence capable of hybridizing to a site including the mRNA's polyA signal sequence and the second primer contains arbitrary sequence. In another approach, the first primer contains arbitrary sequence and the second primer contains sequence capable of hybridizing to a site including the Kozak sequence. In another approach, the first primer contains a sequence that is substantially complementary to the sequence of a mRNA having a known sequence and the second primer contains arbitrary sequence.

Abstract: Reagents and test methods for rapidly and specifically testing maize plants for the presence of T-type cms are provided. The reagent includes a novel nucleic acid segment whose sequence is uniquely arranged in mitochondrial DNA of cms-T maize. The segment, designated TURF 2H3, was cloned and vectors comprising TURF 2H3 provided. Subclones having sequences specific to cms-T mitochondrial DNA and the DNA and deduced amino acid sequences of ORI3, which is unique to T-type cytoplasm, are also provided.

Abstract: A single point mutation in the human lipoprotein lipase gene which results in an A.fwdarw.G nucleotide change at codon 291 (nucleotide 1127) of the lipoprotein lipase gene, and a substitution of serine for the normal asparagine in the lipoprotein lipase gene product is seen with increased frequency in patients with coronary artery disease, and is associated with an increased susceptibility to coronary artery disease, including in particular premature atherosclerosis. This is expressed as a diminished catalytic activity of lipoprotein lipase, lower HDL-cholesterol levels and higher triglyceride levels. Thus, susceptibility of a human individual to premature atherosclerosis and other forms of coronary artery disease can be evaluated byevaluating the sample of DNA for the presence of nucleotides encoding a serine residue as amino acid 291 of the lipoprotein lipase gene product. The presence of a serine residue is indicative of increased susceptibility in the patient.

Abstract: The present invention relates to purified DNA sequences encoding all or a portion of an osteoclast-specific or -related gene products and a method for identifying such sequences. The invention also relates to antibodies directed against an osteoclast-specific or -related gene product. Also claimed are DNA constructs capable of replicating DNA encoding all or a portion of an osteoclast-specific or -related gene product, and DNA constructs capable of directing expression in a host cell of an osteoclast-specific or -related gene product.

Abstract: The complete amino acid and nucleotide sequence for adenylate cyclase stimulating factor is provided, thereby facilitating the synthesis of ACSF in recombinant cell culture. ACSF amino acid sequence variants and ACSF antibodies are provided which are useful in the treatment of humoral hypercalcemia of malignancy or in immunoassays for ACSF. In particular, antibodies capable of binding only the C-terminal domains of ACSF are useful in immunoassays for ACSF which avoid interference by parathyroid hormone. Also provided are novel polypeptides selected from the group of the ACSF basic peptide, the ACSF C-terminal peptide, or the ACSF domain containing both of the basic and C-terminal peptides.

Abstract: Primers specific for enterohemorrhagic Escherichia coli (EHEC) 0157:H7 bacteria have been designed which are useful for detecting the bacteria by polymerase chain reaction methods. The primers were derived from DNA sequences contained within a 60-MDa plasmid which is present in most EHEC. The primers may also be used in combination with primers derived from other sequences of significance, the conserved sequences of Shiga-like toxins I and II and the eaeA gene, in a single simultaneous amplification reaction to specifically identify EHEC serotype 0157.

Abstract: The invention relates to nucleic acid fragments characterized in that they comprise from 8 to 40 nucleotides and in that their sequence is contained either in the DNA coding sequence of the gene for human villin or in any DNA fragment exactly complementary to one of the former and hence containing the same number of deoxynucleotides, or any corresponding RNA fragment containing the same number of ribonucleotides. The invention also relates to the application of these fragments to a procedure for the in vitro detection of the presence of a nucleic acid characteristic of human villin.

Abstract: A method aimed at the quantification of di- and trinucleotide repeat which includes (a) treating a sample containing the nucleic acids of interest to obtain unpaired nucleotide bases spanning the position of the repeats and flanking regions, if the nucleic acids are not already single stranded; (b) contacting the unpaired nucleotide bases with an oligonucleotide primer capable of hybridizing with a stretch of nucleotide bases present in the nucleic acid of interest preferably 3' of the trinucleotide repeats to be quantified, so as to form a duplex between the primer and the nucleic acid of interest; (c) providing means to ensure that the examined nucleic acid and the oligonucleotide primer are confined to a reaction chamber at all further steps; (d) contacting the duplex with a primer extension unit which is capable of base pairing with the first nucleotide base in the core sequence of the repeats, and a template dependent extension enzyme; (e) eliminating non-incorporated primer extension units; (f) contacti

Abstract: Methods are provided for assessing mutations of the APC gene in human tissues and body samples. APC mutations are found in familial adenomatous polyposis patients as well as in sporadic colorectal cancer patients. APC is expressed in most normal tissues. APC is a tumor suppressor.

Abstract: A method for the detection of poor metabolizers of drugs using PCR technology is described comprising a) optionally amplifying a gene coding for an enzyme known to be responsible for the metabolization of drugs, thereby separating it from possible closely related pseudogens,b) amplifying different allelic forms of the gene of step a) and c) detecting the product of step b). Primers for amplification of the genes responsible for the debrisoquine or acetylation phenotype are also disclosed.

Abstract: Novel oligomers are disclosed which have enhanced ability with respect to forming duplexes or triplexes compared with oligomers containing only conventional bases. The oligomers contain the bases 5-(1-propynyl)uracil, 5-(1-propynyl)cytosine or related analogs. The oligomers of the invention are capable of (i) forming triplexes with various target sequences such as virus or oncogene sequences by coupling into the major groove of a target DNA duplex at physiological pH or (ii) forming duplexes by binding to single-stranded DNA or to RNA encoded by target genes. The oligomers of the invention can be constructed to have any desired sequence, provided the sequence normally includes one or more bases that is replaced with the analogs of the invention. Compositions of the invention can be used used for diagnostic purposes in order to detect viruses or disease conditions.

Abstract: A method of marking a liquid and subsequently detecting that the liquid has been marked, which method comprises: adding to the liquid an additive comprising a plurality of particles in an amount no greater than 1 part weight of particles per 10.sup.6 parts weight liquid, the particles comprising signal means to aid their detection and not being visible in the liquid to the naked eye; sampling a portion of the liquid containing said additive, and detecting the presence of particles in the liquid, with the proviso that said signal means does not consist solely of a nucleic acid tag.