Abstract: A hematological assay is described in which the blood coagulation potential of a body fluid is assessed by reacting a sample of the body fluid with an amount of an activator reagent comprising: (a) a predetermined amount of factor Xa or a hematologically equivalent mutant thereof, and (b) a predetermined amount of factor Va, a hematologically equivalent mutant thereof or an enzyme activating endogenous factor V, (c) (optionally) phospholipids. The reagent may be dry (e.g. lyophilised) or in an aqueous solution preferably buffered to a pH from 6 to 10 (preferably 7 to 8), if desired incubating, if necessary inducing coagulation by the addition of one or more coagulation accelerants such as calcium chloride, and establishing a value indicative of the coagulation potential, e.g. by measuring the time to clotting on an optical coagulometer or through use of a chromogenic substrate. It is preferred to use at (b) factor V activator from purified Russell's Viper venom (RVV-V).

Abstract: The invention relates to a process for purifying factor VII and/or activated factor VIIa (F VII/F VIIa) by means of binding to immobilized soluble thromboplastin.

Abstract: The present invention provides methods for inhibiting blood clotting. In general, the methods include adding corn trypsin inhibitor (CTI) to blood or a blood product in an amount sufficient to inhibit the clotting. The CTI can be used alone or in combination with other anti-coagulants. In one aspect, the invention features plasma clotting assays featuring substantially prolonged clotting times. Clotting assays using whole or minimally altered blood are also provided. Further provided are methods for storing blood or blood products at low temperature with the CTI.

Abstract: A process and device are disclosed to determine the activity of enzymes in liquids in a largely automatic manner. The device for carrying out this process has a column (1) with a chromatographic carrier for treating a measurement sample. The carrier is mixed with a substance capable of binding to an enzyme inhibitor present in the measurement sample and that corresponds to at least one enzyme. A measurement sample supply (2) is associated to one end of the column (1). A valve/pump arrangement (7, 11, 14, 15) for filling at least one test tube (5) with a carrier and at least part of the measurement sample is connected downstream of the column (1), in the flow direction of the measurement sample. The carrier is dissociated into cleavage products by the action of the enzyme. The rise in concentration per unit of time of at least one of the cleavage products of the carrier is sensed during an incubation time.

Abstract: Method for screening for the presence of a genetic defect associated with thrombosis and/or poor anticoagulant response to activated protein C (APC). The method is directed at detecting one or more mutations at one or more of the cleavage and/or binding sites for APC of Factor V and/or Factor Va or at Factor VIII and/or Factor VIIIa at either nucleic acid or protein level or both.

Abstract: An immunoassay comprises the steps of: (a) mixing a whole blood sample with sensitized insoluble carrier particles smaller than erythrocytes to cause an immune agglutination reaction; (b) introducing the resulting immune agglutination reaction mixture including agglutinated particles and unagglutinated particles to a flow cell, irradiating the particles passing through the flow cell with laser light, and detecting scattered lights generated thereby; (c) setting a threshold value for distinguishing unagglutinated particles from agglutinated particles and a threshold value for distinguishing the agglutinated particles from blood cells with regard to intensity of the scattered light; and (d) distinguishing and counting the unagglutinated particles, the agglutinated particles and the blood cells from the scattered lights detected in the step (b), in reference to the threshold values set in the step (c).

Abstract: An apparatus for performing a platelet inhibition test on a sample of blood is provided, comprising a plurality of test cells, the cells being adapted for receiving an aliquot portion of the sample, wherein each of the cells comprises an anticoagulant and a clotting activator, and wherein at least one of the cells further comprises a platelet inactivating agent, wherein a clotting time is determined for each of the aliquot portions, and wherein a relative clotting time for each of the aliquot portions comprising the platelet inhibitor is determined as compared to a reference clotting time for a cell containing no platelet inhibitor, wherein the relative clotting times for the cells are determinative of the platelet inhibition of the sample.

Abstract: A method of determining a contact activator for platelet activation and/or clotting activation is provided, comprising placing a predetermined amount of an anticoagulant in each cell of a multicell test cartridge, placing a predetermined amount of a platelet inactivating agent in each cell, and placing a measured amount of contact activator in each cell, the amount of contact activator in each cell differing from the amount in each other cell. An aliquot of a blood sample is added to each cell, and the blood sample aliquot, clotting reagent and platelet inactivating agent are mixed. Each cell sample is allowed to clot, and the clotting time for each cell is measured. The relative clotting times are used to determine the platelet activation and/or clotting activation effect of the clotting activator.

Abstract: Stable compositions containing biotinylated molecules, such as enzymes, are provided. The compositions include a biotinylated biomolecule, a biomolecule protectant, a buffer, a bulking agent selected from one or more water soluble, nonionic polymers and preferably a terminal sterilization protectant. The compositions can be utilized either as aqueous solutions or preferably in dried form, e.g., as a lyophilized, powder cake. They have applicability in any case where avidin/biotin technology is used and are particularly important as compositions containing a thrombin-like enzyme, e.g., for preparation of a fibrin monomer and fibrin monomer-based fibrin sealants.

Abstract: The present invention is directed to methods to rapidly assess the overall coagulant properties of a patient's blood sample by inhibiting the activation of the intrinsic contact activation pathway of coagulation and activating the extrinsic pathway of coagulation. When the sample is whole blood, the resulting clotting time represents the overall coagulant activity of the plasma and cellular components of the blood, which is indicative of existing or impending pathology arising from abnormal coagulability. The invention also provides a method for measuring the risk of a patient for a thrombotic event and for monitoring the effectiveness of procoagulant/anticoagulant therapy. A blood collection apparatus suitable for use in for performing the methods of the invention is also provided.

Abstract: The invention relates to a method of monitoring the effect of a direct of indirect Factor Xa inhibitors comprising the steps of collecting a plasma sample from a patient, adding a solution of Russell's viper venom to the plasma sample and measuring the clotting time or the residual FXa activity chromogenically.

Abstract: A method which may be used to determine haemostatic dysfunction in a patient is carried out by (a) adding a reagent to a test sample, wherein the test sample includes at least a component of a blood sample from a patient; and then (b) measuring the formation of a precipitate due to the reaction of the test sample and the reagent, over time so as to derive a time-dependent measurement profile, the reagent forming a precipitate in the test sample without causing substantial fibrin polymerization.

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: A means for stable preparing a composition, which comprises plasma component derived from human or animals, at least fibrinogen, which is blood coagulation factor, and factor V, is provided, and the composition prepared remains stable even when allowed to stand over one day or longer in a freeze-dried state, in freeze-thawed state, in a refrigerator, or under room temperature. By adding an inhibitor of Factor XIII in an effective amount for achieving the inhibitory effect, compositions comprising human or animal plasma components, at least fibrinogen and Factor V, which are blood coagulation factors, and reagents for measuring extrinsic coagulation factor for blood coagulation function comprising at least thromboplastin, Factor V and fibrinogen can be stabilized during preparation, storage and measurement.

Abstract: Coagulation control compositions suitable for use in connection with PT and/or APTT assays are disclosed along with their methods of preparation and methods of use. Preferred coagulation controls comprise plasma and an anticoagulant having activity for enhancing the activity of antithrombin III (ATIII) or of heparin co-factor II (HCII) against thrombin or against a clotting factor selected from the group consisting of factors IXa, Xa and XIa. The anticoagulant is preferably a glycosaminoglycan such as heparin, a heparin derivative or a heparin analog. The anticoagulant is preferably combined with (1) an abnormal plasma (e.g. activated plasma or factor-deficient plasma) and/or (2) a primate plasma (e.g. human plasma), and a non-primate mammalian plasma (e.g. bovine plasma). In the latter case, the non-primate mammalian plasma is preferably present in the coagulation control composition in an amount of not more than about 12% by volume, relative to total volume.

Abstract: This invention provides methods of inhibiting calcification of a soft tissue (e.g., an artery, a heart valve, an atherosclerotic plaque, a cancer, a kidney, a prostate, skin, muscle, cartilage, viscera, and heart muscle) in a mammal. These methods involve inhibiting osteoclastic bone resorption in said mammal (e.g., a mammal diagnosed as having or at risk for a pathology characterized by calcification of a soft tissue). The inhibition is preferably by administration of a bisphosphonate to the mammal in a concentration sufficient to inhibit bone resorption without inhibiting bone mineralization. The methods of this invention can also be used to mitigate a symptom of atherosclerosis in a mammal. Such methods involve inhibiting osteoclastic bone resorption in the mammal.

Abstract: The invention relates to a method and to a diagnostic agent for detecting hemostasis disturbances, wherein, as a consequence of blood platelet aggregation, clot formation and/or clot dissolution, substances are brought to a distance from each other which permits or prevents an interaction, in particular an energy transfer, between the substances, and the extent of the interaction is measured.

Abstract: A non invasive method for determining the blood coagulation status of a human or an animal is described. F1+2, F1, F2, FpA, D-dimers, F1+2 combined with F1, or F1+2 combined with F2 are measured in saliva, sputum and related biological samples. The respective concentrations are inversely correlated to the time it takes blood to coagulate and the probability of bleeding, and directly correlated to the probability of in-vivo formation of a thrombus or emboli. The method can be used in any coagulation disorder study, including congenital, acquired or drug-induced.

Abstract: The invention relates to oligopeptide derivatives whose C-terminal amino acid is arginine which is linked to the remainder of an electroactive aniline or aminoquinoline derivative by an amide bond. The oligopeptide derivatives are split by enzymes of the class of peptide hydrolases, especially proteinases and their inhibitors, of the coagulation system, the fibrinolytic system and the complement system. These oligopeptide derivatives serve as substrates for quantitatively determining such enzymes, especially thrombin, in complex sample liquids, especially capillary blood. This determination is carried out by measuring the increase in the water-soluble amperogenic aniline or aminoquinoline compound. The oligopeptide derivatives and their salts can be produced according to usual methods in peptide chemistry.

Abstract: Methods for detecting human factor VIII or factor VIII-like polypeptides in and isolating it from solutions such as blood or conditioned media are disclosed, together with reagents suitable for the purpose comprising binding moieties that recognize human factor VIII and/or a factor VIII-like polypeptide and form a binding complex therewith. Preferred polypeptide binding moieties are particularly disclosed.

Abstract: The invention relates to a method and to a diagnostic agent for detecting hemostasis disturbances, wherein, as a consequence of blood platelet aggregation, clot formation and/or clot dissolution, substances are brought to a distance from each other which permits or prevents an interaction, in particular an energy transfer, between the substances, and the extent of the interaction is measured.

Abstract: The present invention generally provides a method of measuring the biological activity of Coagulation Factor VIIa (Factor VIIa). Specifically, the present invention provides a method for directly measuring the activity of Factor VIIa in a plasma sample. More specifically, the present invention provides a method for utilizing Factor VIIa activity as a bio-marker to monitor Factor VIIa inhibition to screen compounds which are able to inhibit the biological activity of Factor VIIa.

Abstract: An anticoagulant for blood cell counting containing at least two kinds of the following antiplatelet agents: theophyllines, adenosines and dipyridamoles. The anticoagulant also contains citric acid, an alkali metal salt thereof or a mixture of citric acid and an alkali metal salt thereof.

Abstract: A sterile method for preparing stable thrombin component from a single donor's plasma in which the thrombin component and the clotting and adhesive proteins component are harvested simultaneously 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: Blood is tested for clot lysis conditions such as natural lytic capabilities, the effect of previously administered thrombolytic and anti-thrombolytic agents, and dose responses thereto, by forming a clot in a sample of blood, lysing the clot, and measuring the elapsed time period from initial clot formation to clot lysis, all while continuously evaluating the blood sample. Thrombolytic agents include streptokinase, urokinase and recombinant tissue plasminogen activator. Plasmin and plasminogen activator inhibitors, clot activating agents (e.g. kaolin), and agents to deactivate anticoagulants (e.g. heparinase) may also be used as reagents during testing.

Abstract: A method is provided for the localized intravascular administration of a fibrinolytic metalloproteinase to a human subject in amounts that are both safe and effective to lyse an occluding fibrin-containing blood clot, while also avoiding the neutralizing effects of &agr;2-macroglobulin in the circulating blood.

Abstract: The invention relates to a method for detecting in a body fluid sample at least one blood coagulation activity marker that reflects e blood coagulation activity of an individual. By correlating the amount or concentration of the blood coagulation activity marker present e.g. in a urine sample, it is possible to monitor the blood coagulation activity of a patient following surgery without having to obtain a blood sample from said patient.

Abstract: The invention provides monoclonal antibodies which are reactive with fibrin(ogen) degradation products (FDPS) generated by matrix metalloproteinases (MMPs), such as MMP-3, MMP-7, and membrane-type 1, MT1-MMP proteolysis of fibrinogen and cross-linked fibrin (and not plasmin or other proteases). Monoclonal antibodies of the invention include, but are not limited to, H5, F2, 90/2, 90/5, B4, 13, C6, A6, G6, E6, 197-1 and 197-2. This invention also provides methods of using monoclonal antibodies for detection of FDPs generated by proteolysis of fibrin(ogen) by MMPs, such as MMP-3, MMP-7, and MT1-MMP. In addition, the present invention provides methods for diagnosing rheumatoid arthritis, osteoarthritis, and synovitides, as well as angiogenesis, atherosclerosis, renal diseases, malignancy and inflammation.

Abstract: Provided are a monoclonal antibody making it possible to detect a native fibrin monomer, which is produced at the initial state of blood coagulation, and soluble fibrin; a hybridoma; and an immunoassay for detecting the initial stage of blood coagulation with high sensitivity, quickly, using the monoclonal antibody. Using a fibrinogen analog in blood as an immune source, cell fusion is carried out to prepare a monoclonal antibody which is not reactive with fibrinogen and is specifically and simultaneously reactive with a native fibrin monomer (that is, a fibrin monomer which is present in a body fluid, in particular in blood, and is not solubilized) and soluble fibrin. The fibrin monomer analog is preferably fibrinogen treated with bathroxobin, which is a snake venom.

Abstract: An activated partial thromboplastin time (APTT) test is described which employs fresh, whole blood that has not been anticoagulated. The method employs a disposable cuvette having specific reagents placed therein. A sample of whole blood is introduced into the cuvette which is drawn into at least one conduit by a pneumatic pump. Initially, the calcium in the sample is chelated at a first reaction site in the conduit, and thereafter is moved through the conduit to a second reaction site where the sample is coagulated by an APTT reagent. After a predetermined incubation time, the sample is moved to a third reaction site where it is recalcified. Clotting time is measured from the time that the sample is recalcified to the time that clot formation is visibly detected.

Abstract: A coagulation test for determining the activated clotting time (ACT) of blood in the presence of heparin that produces test results that are substantially insensitive to the drug aprotinin. The activator is formulated to be a combination of celite and bentonite. The ACT results obtained with this formulation are similar to celite ACT tests on heparinized blood while simultaneously being unaffected by aprotinin. Additionally, a method for quantifying the aprotinin effect of different ACT formulations is disclosed.

Abstract: The invention is a method, reagent, test cartridge and device for the determination of fibrinogen in a sample, such as undiluted blood and blood plasma. The method utilizes a reagent comprising thrombin and a thrombin inhibitor that slows the enzymatic conversion of fibrinogen to fibrin, and thus slows clotting in samples. This is particularly useful for determining fibrinogen in undiluted blood and blood plasma. Fibrinogen assay reagents comprising at least one thrombin inhibitor and thrombin, and test cartridges containing the reagents for automated analysis of fibrinogen are disclosed, along with automated devices using said reagent and test cartridges for the devices.

Abstract: The invention relates to compounds inhibiting the activation of FX to FXa by TF/FVIIa. The compounds are anticoagulants. The invention also relates to a method of identifying a drug candidate.

Abstract: In vitro methods for qualitative screening and/or quantitative determination of the functional activity of components of the Protein C anticoagulant pathway of blood coagulation are described. The methods entail measuring the conversion rate of a substrate by an enzyme, the activity of which is related to the Protein C anticoagulant activity, in a blood sample of a human comprising coagulation factors and said substrate, after at least partial activation of coagulation through the intrinsic, extrinsic or common pathway and triggering coagulation by adding calcium ions; and comparing said conversion rate with the conversion rate of a normal human blood sample determined in the same way. The methods include the addition of additional metal ions to the sample to enhance activity, sensitivity and resolution. Kits and reagents for use in the methods are also provided.

Abstract: A method for determining the existence and the amount of soluble fibrin contained in a specimen fluid is provided. The method includes the steps of precipitating soluble fibrin out of the opaque specimen fluid, aggregating the soluble fibrin precipitates in a limited region of a transparent container so as to render the precipitates optically detectable in the opaque specimen fluid, and optically detecting the precipitates. The amount of soluble fibrin may be determined by measuring the time from the addition of the precipitating regent to the detection of the soluble fibrin precipitates. Methods of the present invention allow one to measure soluble fibrin in whole blood, and therefore render the test useful in the operating room under conditions of major surgery and in the presence of severe trauma wherein DIC is likely to supervene.

Abstract: The invention provides a one-stage method of measuring antithrombin (AT) activity in a sample. In the method, a diluted sample is mixed with AT-deficient plasma containing intrinsic coagulation enzymes, an AT augmenting compound such as heparin, a phospholipid and an activator of the contact phase of the intrinsic coagulation pathway. Following addition of calcium ions, coagulation time is measured and compared to a reference standard to determine the level of AT activity in the sample.

Abstract: A method for determining coagulation parameters which is safe, convenient, applicable to existing automated analyzers and reliable is disclosed. The said method for the determination utilizes an antibody against the E region of fibrinogen, fibrinogen degradation product D dimer or the like, and is enable to determine the coagulation parameter in fibrin coagulation system accompanied by limited degradation of fibrinogen with thrombin without clotting.

Abstract: Disclosed is The method of in vitro testing of the state of the blood coagulation system. The Accelerated Whole Blood Clotting Time (A. W. B. C. T.) reflects the degree of over or under activity of the coagulation system of the blood.

Abstract: A method for screening for anti-thrombotic/anti-platelet agents is provided where the method is based on inhibition of the GP Ib, PAR-1 and/or PAR-4 pathways by the potential antithrombotic/anti-platelet agent.

Abstract: The method relates to a method for detecting disorders in the protein C system, and in particular for determining the activated blood coagulation factor V with increased stability with respect to decomposition by activated protein C.

Abstract: Histamine may be quantitatively measured by performing the following steps. First, an oocyte that expresses histamine receptors is held in a recess formed at the bottom of a vessel. Then, first and second electrodes are inserted into the oocyte. Subsequently, the membrane potential of the oocyte is measured by using the first electrode to stabilize this membrane potential at a predetermined level by driving a current through the second electrode using circuitry for clamping the membrane potential of the oocyte. A sample is then infused into a fine reacting tube having an antigen immobilized on its inner surface together with some buffer solution to promote a histamine releasing reaction. The solution containing histamines that is released in the fine reacting tube is transferred to the vessel to make contact with the oocyte in the vessel.

Abstract: Providing an evacuated blood collection tube for blood collection rapidly, whereby serum of high purity can be isolated in a rapid and simple manner with no occurrence of fibrin deposition. Beads each coated with a blood coagulation-promoting enzyme comprising thrombin and batroxobin are contained in the evacuated blood collection tube. Preferably, thrombin is used in an amount of 0.1 to 3 IU and batroxobin is used in an amount of 0.25 to 2 BU, per 1 ml of collected blood.

Abstract: The existence and the extent of periodontal disease can be diagnosed by measuring plasminogen activator inhibitor 2 (PAI-2) and/or tissue plasminogen activator (t-PA) levels in gingival crevicular fluid (GCF). Levels of PAI-2 and t-PA in GCF rise sharply in the context of periodontal disease, and they also correlate with the severity of disease at different sites in the same patient.

Abstract: The present invention relates to a method of determining the enzymatic activity of blood coagulation factor XIII by using purified fibrin monomer as a substrate of this enzyme. The enzymatic activity is determined by detecting the degree of cross-linking of fibrin monomer formed by the blood coagulation factor XIII and the fibrin monomer free of blood coagulation factor XIII is obtained by washing the preparation with citric acid solution. The present method can be effectively used for the validation of the other enzymatic assay methods for the blood coagulation factor XIII as well as the studies for the characterization of the blood coagulation factor XIII.

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 of measuring erythrocyte sedimentation rate (ESR), plasma viscosity, or plasma fibrinogen of a blood sample is provided. The method involves treating the blood sample with a suitable anti-coagulant, placing the treated sample in a micro-haemocrit tube and centrifuging the sample at, for example, 500 rpm or greater while continually monitoring the sample using an IR light source and detector to follow appropriate separation band development during centrifugation. Thus, continual monitoring of the sample during the centrifugation process is possible. Preferably, the sample is horizontal throughout the centrifugation process.

Abstract: The present invention provides methods for inhibiting blood clotting. In general, the methods include adding corn trypsin inhibitor (CTI) to blood or a blood product in an amount sufficient to inhibit the clotting. The CTI can be used alone or in combination with other anti-coagulants. In one aspect, the invention features plasma clotting assays featuring substantially prolonged clotting times. Clotting assays using whole or minimally altered blood are also provided. Further provided are methods for storing blood or blood products at low temperature with the CTI.

Abstract: In vitro methods for qualitative screening and quantitative determination of the functional activity of components of the Protein C anticoagulant pathway of the blood coagulation system are provided. The methods entail providing a blood sample to be analyzed, activating the coagulation cascade, triggering coagulation by adding calcium ions, adding metal ions selected from the group consisting of divalent metal ions and monovalent copper ions at a concentration that enhances the anticoagulant activity of the Protein C anticoagulant pathway, adding an exogenous substrate for an enzyme related to Protein C anticoagulant activity, measuring the conversion rate of a the exogenous substrate by the enzyme, and comparing the conversion rate of the substrate in blood sample to be analyzed with the conversion rate the substrate in a normal blood sample.

Abstract: The present invention relates to a process for increasing the FVII sensitivity of a thromboplastin reagent by means of heat treatment.

Abstract: The present invention provides methods for inhibiting blood clotting. In general, the methods include adding corn trypsin inhibitor (CTI) to blood or a blood product in an amount sufficient to inhibit the clotting. The CTI can be used alone or in combination with other anti-coagulants. In one aspect, the invention features plasma clotting assays featuring substantially prolonged clotting times. Clotting assays using whole or minimally altered blood are also provided. Further provided are methods for storing blood or blood products at low temperature with the CTI.