Abstract: Processes are disclosed using the depolymerization of a nucleic acid hybrid to qualitatively and quantitatively analyze for the presence of predetermined nucleic acid target sequences using a multiplex assay format. Applications of those processes include the detection of single nucleotide polymorphisms, identification of single base changes, speciation, genotyping, medical marker diagnostics, and the like.

Abstract: The present invention relates to novel antithrombotic variants of thrombin or fragments thereof that are capable of proteolytically activating protein C, but which are substantially free of fibrinogen cleavage activity. The present invention further relates to variant polypeptidess that may be cleaved to yield active thrombin variants. The present invention also relates to methods of inhibiting thrombus formation in an animal or human subject by delivering an antithrombotic variant thrombin of the present invention to the blood of the subject. The present invention relates also to methods that use the novel variant thrombins for determining the level of protein C activation in a blood sample, or the thrombogenic potential of a patient.

Abstract: The invention provides novel polynucleotides and polypeptides encoded by such polynucleotides and mutants or variants thereof that correspond to a novel human prothrombinase-like polypeptide. These polynucleotides comprise nucleic acid sequences isolated from cDNA library from human ovary (Hyseq clone identification number 7735656). Other aspects of the invention include vectors containing processes for producing novel human prothrombinase-like polypeptides, and antibodies specific for such polypeptides.

Abstract: A method for the refinement of a thrombin substrate, characterized by comprising therein a step of affinity chromatography formed by using an affinity adsorbent having as a ligand a thrombin or thrombin analogue deprived of nucleophilicity (activity) without causing stereostructure to the linkage with a substrate is disclosed.

Abstract: A method of producing a pharmaceutical preparation comprising fibronectin and fibrinogen is disclosed. The method involves admixing into a starting solution of fibrinogen and fibronectin, in a single step, a precipitating composition comprising a polyalkylene glycol and at least one of glycine and &bgr;-alanine which forms a precipitate. Next, the precipitate is collected and a pharmaceutical preparation is prepared from the precipitate. The pharmaceutical preparation has a fibronectin:fibrinogen ratio from about 0.02 to about 0.2.

Abstract: The present invention provides compositions effective in decontaminating either biological pathogens or both chemical and biological pathogens. These compositions are particularly suitable for the decontamination of biological warfare agents or both chemical and biological warfare agents The compositions comprise generally a blend of biocides, and may additionally comprise a protein and an enzyme. Further, the composition is contained in a buffered foam forming material for ease in distribution The compositions are nontoxic, noncorrosive and nonflammable.

Abstract: The present invention provides novel nucleic acids, novel polypeptide sequences encoded by these nucleic acids and uses thereof.

Abstract: A method, kit, system and reagent for performing coagulation assays with higher sensitivity and greater dynamic range is provided which involves the use of one or more metal compounds that interact with calcium binding sites in the blood coagulation cascade, particularly lanthanide compounds, manganese compounds and magnesium compounds. A Protein C reagent, kit, and assay method is also provided using the same type of metal compounds to provide greater detection sensitivity and dynamic range between samples.

Abstract: The invention provides a kit for preparing a fibrin sealant either (A) comprising: (a) a fibrin monomer preparation; (b) a stabilizing preparation containing a clot-preserving effective amount of a fibrinolysis-inhibiting protein; and (c) a non-enzymatic polymerizing agent preparation effective to convert the fibrin monomer preparation into a fibrin clot; or (B) comprising: (a′) a fibrin monomer preparation comprising a fibrin monomer and a clot-preserving effective amount of a of a fibrinolysis-inhibiting protein; and (c′) a polymerizing agent preparation effective to convert the fibrin monomer preparation into a fibrin clot.

Abstract: In accordance with the present invention a novel fibrin polymer structure is disclosed. The novel fibrin polymer structure useful, for example, as a surgical sealant, is comprised of a plurality of discrete, droplets of polymerizing or polymerized fibrin each encapsulated by a “skin” of fibrin polymer. These fibrin-skin encapsulated droplets are applied so as to be built up one upon the other, layer by layer, to form an integral sealant structure. The cumulative effect of the encapsulating skins of those droplets which form the sealant surface is a surface skin which unexpectedly resists cell penetration but enhances cell migration across the surface. The sealant structure of the present invention can be prepared by spray delivery of fibrin polymer forming materials wherein the time required for the materials to commence polymerizing after mixing is less than or equal to the transit time of said materials from the applicator tip to the target surface.

Abstract: According to the invention there is provided the use of melagatran, or a pharmaceutically-acceptable derivative thereof, for the manufacture of a medicament for the treatment of ischemic disorders in patients having, or at risk of, non-valvular atrial fibrillation.

Abstract: New forms of ecarin, a procoagulant protein from Echis carinatus venom, are described, as are polynucleotides encoding the new proteins, methods for production of the new proteins, and methods for activation of prothrombin using the new proteins. The new ecarins comprise a serine at position 396 of the protein. The new proteins may be used for activation of prothrombin, and are particularly useful for the production of recombinant thrombin.

Abstract: A method of purifying antithrombin III (AT III) from a starting material containing an AT III/heparin complex or an AT III/heparinoid complex is disclosed. First, the method comprises adsorbing the AT III/heparin complex or the AT III/heparinoid complex on an anion exchanger material. Second, the method involves separating the AT III from the adsorbed AT III/heparin complex or an AT III/heparinoid complex by elution with a buffer having a pH ranging from 8.5 to 10.5 and a conductivity between 10 and 60 mS.

Abstract: A process is disclosed for the baseline separation of the &agr; and &bgr; variants of antithrombin III AT-III. The process involves subjecting a sample comprising AT III to cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC). The CD-MEKC is performed with a buffer comprising boric acid, an anionic tensid, &bgr;-cyclodextrin, and an aliphatic diamine at a basic pH. The CD-MEKC results in the separation of the AT III into AT III &agr; and ATIII &bgr;. The anionic tensid can be sodium dodecyl sulfate and the aliphatic diamine can be 1, 5-diaminopentane.

Abstract: The invention relates to the treatment of subjects for the purpose inhibiting vaso-occlusive events, including thrombosis and embolism, by administering agents which reduce the number of circulating platelets to low or below normal levels. Methods and pharmaceutical preparations comprising such agents are provided.

Abstract: The application relates to a method for determining the anticoagulatory potential of a sample by adding thrombomodulin and thromboplastin in a coagulation test.

Abstract: Human tissue plasminogen activator (t-PA) is produced in useful quantities using recombinant DNA techniques. The invention disclosed thus enables the production of t-PA free of contaminants with which it is ordinarily associated in its native cellular environment. Methods, expression vehicles and various host cells useful in its production are also disclosed.

Abstract: A sterile method for preparing stable thrombin component from a single donor's plasma in which the thrombin component is harvested simultaneously from the clotting and adhesive proteins component from the same donor plasma in less than one hour. The combined components provide an improved biological hemostatic agent and tissue sealant by virtue of its freedom from the risk of contaminating viruses or bacteria from allogenic human or bovine blood sources. The thrombin provides polymerization of the clotting and adhesive proteins in less than five seconds, and is sufficiently stable to provide that fast clotting over a six hour period. Further, the clotting times can be predictably lengthened by diluting the thrombin with saline.

Abstract: The subject invention relates to fibrin sealants. More specifically, the subject invention relates to the use of a fibrin sealant wherein a composition comprising fibrin monomer or a composition comprising noncrosslinked fibrin is utilized as a component of the fibrin sealant.

Abstract: A method for the production of thrombin from pure prothrombin is provided by converting pure prothrombin by treating pure prothrombin with sodium citrate in the absence of additional coagulation factors. Preferrably, a solution is used containing about 30% to about 40% sodium citrate.

Abstract: The invention relates to novel protease-inhibitors which are obtainable from leeches. It also relates to uses thereof, for instance as a medicament, thus pharmaceutical preparations are provided, as are derivatives, mutants, genes encoding, vectors comprising and cells provided with such genes and/or vectors. In particular the invention relates to a family of proteinaceous protease-inhibitors having a molecular weight of about 5.5 kD and the following primary sequences: DDNCGGKVCSKGQLCHDGHCECTPIRCLIFCPNGFAVDENGCELPCSCKHQ, DDDCGGQVCSKGQLCVDGQCKCTPIRCRIYCPKGFEVDENGCELPCTCLQ and DGNCGGQVCSKGQLCVDGQCKCTPIRCRIYCPKGFEVDENGCELPCTCLQ. This invention also relates to HIV-inhibitors and other therapeutically interesting, low molecular weight, and low antigenic substances from leeches.

Abstract: A method for the preparation of thrombin from a prothrombin-containing material is characterised in that the prothrombin-containing material is an activated prothrombin complex concentrate ActPCC fraction, which is contacted with divalent cations to convert the prothrombin to thrombin.

Abstract: Novel polypeptides (NPs) are provided which are capable of protein C activation without significant fibrinogen clotting activity, and vice versa. NPs having enhanced protein C activating properties in relation to fibrinogen clotting are useful in particular as anticoagulants and in screening for substances that agonize or antagonize this property and in diagnostic procedures to determine the status of patients' activated protein C-mediated anticoagulant pathway. Procoagulant NPs are useful to promote clotting in the course of therapy of solid tumors, as an impregnate for bandages, or in diagnostic assays. The NPs are produced in recombinant cell culture or by in vitro methods.

Abstract: The present invention provides methods and compositions relating to synthetic test stains comprising isolated fibrin and/or fibrin precursors and blood plasma proteins. Kits for the preparation of synthetic test stains containing an isolated fibrin precursor and an initiator for converting the fibrin precursor to fibrin are also provided. In addition, methods for checking the efficiency of cleaning processes, and kits for carrying out these methods are provided.

Abstract: The subject invention relates to fibrin sealants. More specifically, the subject invention relates to the use of a fibrin sealant wherein a composition comprising fibrin monomer or a composition comprising noncrosslinked fibrin is utilized as a component of the fibrin sealant.

Abstract: The invention concerns thrombin muteins which have the following differences in their sequences as compared with natural thrombin: (a) the exchange of a Gly after Ala, the Gly being in the sequence environment Tyr-Gly-Phe and having position 558 in natural human thrombin (SEQ ID No. 14); (b) at least one exchange or deletion of the following radicals: (b1) His, located in the sequence environment Ala-His-Cys and having position 363 in natural human thrombin (SEQ ID No. 14); (b2) Asp, located in the sequence environment Arg-Asp-Ile and having position 419 in natural human thrombin (SEQ ID No. 14); (b3) Ser, located in the sequence environment Asp-Ser-Gly and having position 525 in natural human thrombin (SEQ ID No. 14). The invention further concerns the use of these muteins as antidotes.

Abstract: The subject invention relates to fibrin sealants. More specifically, the subject invention relates to the use of a fibrin sealant wherein a composition comprising fibrin monomer or a composition comprising noncrosslinked fibrin is utilized as a component of the fibrin sealant.

Abstract: This invention relates to a process for preparing biological glue components from a plasma pool which combines high recovery, quality product and viral safety. In first instance, a triple viral inactivated product comprising fibrogen, fibronectin and FXIII is obtained by treating a concentrate thereof first with a viricide solvent/detergent solution, second with viral nanofiltration, and third with heat. The recovery of a good quality product is not compromised by the process of the invention. In second instance, the same steps are reproduced for obtaining a triple viral safe thrombin product. In that case, a known proprietary process has been improved to increase the recovery of active thrombin by about two fold. One of the steps which increase the yield of thrombin is the dilution of the prothrombin solution with water 4 volumes to 1 volume prothrombin prior to acid precipitation.

Abstract: Proteins which have activity as anticoagulants and/or serine protease inhibitors and have at least one NAP domain and are described. Certain of these proteins have factor Xa inhibitory activity and others have activity as inhibitors of factor VIIa/TF. These proteins can be isolated from natural sources as nematodes, chemically synthesized or made by recombinant methods using various DNA expression systems.

Abstract: The present invention relates to the use of a class of genes called oil body protein genes that have unique features. The discovery of these features allowed the invention of methods for the production of recombinant proteins wherein a protein of interest can be easily separated from other host cell components. The invention is further exemplified by methods for exploitation of the unique characteristics of the oil body proteins and oil body genes for expression of polypeptides of interest in many organisms, particularly plant seeds. Said polypeptides may include but are not limited to: seed storage proteins, enzymes, bioactive peptides, antibodies and the like. The invention can also be modified to recover recombinant polypeptides fused to oil body proteins from non-plant host cells. Additionally the invention provides a method of using recombinant proteins associated with seed oil bodies released during seed germination for expression of polypeptides that afford protection to seedlings from pathogens.

Abstract: The present invention provides a production method of thrombin which can convert prothrombin into thrombin highly efficiently even in the absence of thromboplastin and blood plasma, easily obtain a starting material for use in the conversion of prothrombin into thrombin, and prepare and purify thrombin in an industrial scale. It is characterized in treating a prothrombin-containing aqueous solution with a Ca salt at 0 to 15.degree. C.

Abstract: A method for a synthesis of anhydrothrombin is provided which features a short process, an easy procedure, and high yields.The method comprises(A) a step of causing an active serine residue site of thrombin to react with an inhibitor,(B) a step of performing an alkali treatment at a pH of not less than 11, and(C) a step of performing an operation of recovery, and carries out these steps sequentially in the order mentioned, and is characterized by causing at least the step of performing the operation of recovery to proceed in the presence of at least one compound selected from the group consisting of polyhydric alcohols and saccharides, and a salt or an amphoteric electrolyte.

Abstract: An improved method of assaying undiluted whole blood or undiluted plasma for prothrombin is disclosed. The improved method utilizes carinactivase-1 and calcium ions, and eliminates the need for a prothrombin-deficient plasma as a reference plasma.

Abstract: It was found that the use of a phospholipid dependent prothrombin activator purified from the venom of snakes belonging to the Elapidae family, especially members of the Oxyuranus and Psuedonaja genera is most useful in tests for the determination of Lupus Anticoagulant. Based on this, several clotting, chromogenic and immunochromogenic tests have been developed.

Abstract: A pharmaceutical preparation contains protein C and a thrombolytically active substance that does not activate protein C. This preparation prevents reocclusion usually occurring in the course of thrombolysis therapy.

Abstract: A method for activating prothrombin to thrombin is presented which comprises treating an aqueous solution containing prothrombin with polyethylene glycol in the presence or absence of the calcium salt. The method enables conversion of prothrombin to thrombin in the absence of thromboplastin and allows for preparation of thrombin on an industrially large scale from easily available starting materials.

Abstract: The invention relates to fibrin sealants. More specifically, the invention relates to the preparation of a fibrin polymer wherein the polymer is substantially free of the enzyme.

Abstract: Proteolytically inactive thrombin mutated at positions 43 and/or 99 and/or 205, whereby said mutation renders thrombin an antagonist of thrombin receptor.

Abstract: The subject invention relates to the use of thrombin in a medical procedure in an animal. More specifically, the subject invention relates to such use of thrombin wherein the thrombin is a thrombin blood fraction is autologous to that animal.

Abstract: The subject invention relates to the use of thrombin in a medical procedure in an animal. More specifically, the subject invention relates to such use of thrombin wherein the thrombin is a thrombin blood fraction, is autologous to that animal.

Abstract: Proenzymes or proforms of blood factors can be cleaved in the presence of a detergent or a chaotropic substance to produce active blood factors selected from the group consisting of Factor Va, Factor VIIa, Factor VIIIa, Factor IXa, Factor Xa, Factor XIa, Factor XIIa, Factor XIIIa and activated protein C. The chaotropic substance can be urea, guanidinium hydrochloride or a thiocyanate salt. Under these conditions, proteolytic activation occurs in a controlled and restricted manner. Consequently, it is possible to isolate high yields of active blood factor, while minimizing the production of inactive degradation products. Immobilization of the proenzyme or proform on a solid support prior to activation facilitates the separation of active blood factor from the proenzyme or proform and inactive peptide fragments.

Abstract: The present invention provides a method for determining thrombotic risk in an individual. The method involves determining the activity of Protein C and Protein S in the plasma of the individual thought to be at thrombotic risk by adding to a plasma sample obtained from the individual (i) a first reagent in an amount sufficient to induce or activate coagulation in the plasma, (ii) a second reagent which activates endogenous protein C in the plasma, and (iii) a third reagent comprising calcium salts, phospholipids or tissue thromboplastin, or a combination thereof. To a second plasma sample from the same subject is added a reagent which induces or activates coagulation, and a buffer or other material which does not activate protein C, and a third reagent as described above. The time, rate or both, necessary for the conversion of endogenous fibrinogen to fibrin in both the first and second samples is measured.

Abstract: The disclosed Factor V Ratio (FVR) screening blood assay (read as "factor five ratio") and kits for the conduct thereof, identify individuals that possess a specific genetic defect, known as the Factor V Leiden defect, or other genetic or acquired Factor V defect, that makes those individuals susceptible to venous thromboembolism. In this test the Factor V activity of a blood plasma sample exposed to activated Protein C (APC) is compared to the Factor V activity of a similar sample in the absence of APC, after both samples had been treated with an activating agent. The ratio between the Factor V activity level without APC and the Factor V activity level with APC, identifies individuals at risk of a thrombotic disorder due to a Factor V defect and differentiates between individuals with a heterozygous defect and individuals with a homozygous defect.

Abstract: The subject invention relates to methods and compositions using nondynamic fibrin monomer which is selectively converted to a fibrin polymer which serves as the fibrin sealant.

Abstract: The invention disclosed relates to thrombolytic combination therapy by the administration of an agent capable of activating native plasminogen, and a non-native plasminogen analogue which is cleavable by native thrombin to generate plasmin activity.

Abstract: The invention relates to serine protease variants derived from precursor serine proteases via recombinant and/or chemical methods to form protease variants having improved peptide ligase activity. The invention also includes novel ligation substrates which in combination with the serine protease variants and a second ligation substrate are capable of forming a ligation product. The invention also relates to methods for forming such ligation products and the products formed thereby.

Abstract: A thrombin of human or animal origin is free of infectious agents and is produced by activation of prothrombin subjected to a heat treatment for the inactivation of infectious agents.

Abstract: The present invention relates to a method to determine the concentration of anticoagulants as a function of the inhibition of thrombin formation in an assay mixture. The assay mixture is composed of two reagents and the diluted plasma sample. The diluted plasma sample containing an anticoagulatory substance is mixed with a clotting factor reagent, which is a combination of an excess amount of purified coagulation factors of the endogenous system, phospholipids and a weak thrombin inhibitor, which can be potentiated by an anticoagulatory substance. Then a coagulation cascade activator and calcium chloride, an activator reagent, is added to the mixture and formed thrombin is measured after a chosen incubation time. A proportionality between the inhibition of the thrombin formation and the anticoagulatory active components allows for a determination of the concentration of anticoagulatory active components in the sample.

Abstract: Novel compounds of the formula I ##STR1## in which R.sup.1 and R.sup.2 are hydrogen, C.sub.1 -C.sub.4 alkyl or are linked to form C.sub.3 -C.sub.7 cycloalkyl and R.sup.3, R.sup.4 and R.sup.5 are the same or different and are hydrogen, C.sub.1 -C.sub.4 alkyl, hydroxy, OR.sup.6, SR.sup.6, halogen, NR.sup.7 R.sup.8, NO.sub.2, CN, CONR.sup.7 R.sup.8 or CO.sub.2 R.sup.9 wherein R.sup.6 is C.sub.1 -C.sub.4 alkyl or C.sub.7 -C.sub.10 aralkyl and R.sup.7, R.sup.8 and R.sup.9 are the same or different and are hydrogen, C.sub.1 -C.sub.4 alkyl or C.sub.7 -C.sub.10 aralkyl or R.sup.7 and R.sup.8 together with the nitrogen atom to which they are bound form 5 or 6 membered azacycloalkyl or oxazacycloalkyl; Arg is arginine ?NH--CH(CH.sub.2 CH.sub.2 CH.sub.2 NH--C(.dbd.NH)--NH.sub.2)--CO!; X is methine CH or nitrogen; n is an integer from 0 to 7; L is a peptide linker, and H is the carboxy terminal end of hirudin, and salts thereof, are provided.

Abstract: The present invention provides a method for activating prothrombin to thrombin in a prothrombin complex composition comprising at least prothrombin, Factors V, VII, IX, and X, and phospholipid, comprising the steps of: (a) cold-incubating the prothrombin complex composition with 2-15 mM of Ca.sup.2+ ions at a pH of 6.5-8.0 and at a temperature of 2.degree.-8.degree. C. until Factor VII contained in the composition is activated; and (b) incubating the cold-incubated composition at a pH of 6.5-8.0 and at a temperature of 10.degree.-25.degree. C. for a period of time sufficient to activate prothrombin contained in the composition to thrombin.