Abstract: Disclosed is a method that includes steps of identifying conditions each associated with a low level activation of the coagulation response; conducting a clinical evaluation of a patient to determine a possibility that the patient has one of the conditions; if the clinical evaluation indicates the possibility that the patient has one of the conditions, providing a panel of different quantitative blood tests each capable of providing one of an abnormal result indicative of a low level activation of the coagulation response, and a normal result; performing the panel of different quantitative blood tests on the patient; obtaining a result for each of the blood tests; observing the results; and if at least two of the results are abnormal, using the abnormal results to assist in diagnosing the patient with the one of the conditions.

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 relates to a method for measuring the aggregation or agglutination of platelets, where a reaction mixture is mixed in a first reaction phase, and is mixed less vigorously or not at all in a second reaction phase following the first, and the measurement is preferably carried out in the second reaction phase.

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: Specific amino acid loci of human factor VIII interact with inhibitory antibodies of hemophilia patients after being treated with factor VIII. Modified factor VIII is disclosed in which the amino acid sequence is changed by a substitution at one or more of the specific loci. The modified factor VIII is useful for hemophiliacs, either to avoid or prevent the action of inhibitory antibodies.

Abstract: A self-contained analyzer quantitatively detects a clotting event in a biological fluid. The device includes a base having an open reservoir for receiving the biological fluid. A carrier for a dried activator reagent is located in or adjacent to the open reservoir. The biological fluid reconstitutes the dried reagent and starts the clotting process. During the clotting process the fluid travels through a capillary flow channel lined with a precision woven fabric. When clotting is completed, the woven fabric stops the flow of the fluid. A colored bar formed by the fluid flowing through the capillary channel indicates the length of time required for clotting of the sample.

Abstract: The present invention has determined that exogenously added glycosylceramide (GlcCer) and other neutral glycolipids such as the homologous Glc-containing globotriaosylceramide (Gb3Cer), dose-dependently prolonged clotting times of normal plasma in the presence but not absence of APC:protein S, indicating GlcCer or Gb3Cer can enhance protein C pathway anticoagulant activity. In studies using purified proteins, inactivation of factor Va by APC:protein S was enhanced by GlcCer alone and by GlcCer, globotriaosylceramide, lactosylceramide, and galactosylceramide in multicomponent vesicles containing phosphatidylserine and phosphatidylcholine. Thus, the present invention provides neutral glycolipids such as GlcCer and Gb3Cer, as anticoagulant cofactors that contribute to the antithrombotic activity of the protein C pathway. The present invention has also determined that a deficiency of plasma GlcCer is a risk factor for thrombosis.

Abstract: A method, kit, system and reagent for measuring low molecular weight heparin in a whole blood sample is provided which involves the use of a Factor Xa activator, such as Russell's Viper Venom, as the coagulation assay initiator.

Abstract: This invention is a disposable cartridge for use at the patient side to perform traditional coagulation assays on fresh whole blood or blood derivative samples. The cartridge, in use with an electronic analyzer allows a fluid sample to be metered and quantitatively mixed with reagents which activate the coagulation cascade. An artificial substrate for thrombin, the enzyme whose action results in clot formation is also provided. Clot formation is subsequently detected using a microfabricated sensor also housed within the cartridge which detects electrochemically the product of the thrombin reaction upon the synthetic substrate.

Abstract: A method is disclosed for determining if a patient is hypercoagulable, hypocoagulable or normal. The test involves providing a test sample from the patient and initiating coagulation in the sample in the presence of an activator, which is added to the sample in an amount which will result in intrinsic tenase-dependent fibrin. Then the formation of the intrinsic tenase-dependent fibrin polymerization is monitored over time so as to derive a time-dependent profile, with the results of the fibrin polymerization monitoring determining whether the patient is hypercoagulable, normal or hypocoagulable. The coagulation activator is added in an amount that triggers a thrombin explosion that is dependent on the propagation phase and amplification pathways. In this way, a single assay can assess the hemostatic potential of a sample.

Abstract: A process is provided for the assessment of an active proteolytic enzyme in a blood or another biological fluid sample possibly comprising a complex of said proteolytic enzyme and &agr;2-macroglobulin, wherein said sample is contacted with a substrate comprising a molecule of sufficient size coupled to a signal-substrate, said signal-substrate comprising a detectable leaving group, wherein said substrate is hydrolysed by said proteolytic enzyme but not by said complex. The proteolytic enzyme is preferably selected from the group consisting of thrombin, activated clotting factor, activated fibrinolytic factor, and activated component of the complement system. Of these, thrombin is most preferred. The molecules of sufficient size are preferably water soluble and selected from the group consisting of inert protein, preferably ovalbumin, polysaccharide, and synthetic polymer. The size of these molecules is such that they will not fit into the cavity of the &agr;2M molecules.

Abstract: A reagent kit capable of distinguishing between a blood sample containing lupus anticoagulant and blood samples from individuals having other anticoagulant diseases is disclosed. The reagent kit comprises two coagulation time reagents containing phosphatidylserine in a different phosphatidylserine content ratio to the total content of phospholipids from each other, threreby giving different coagulation times with use of corresponding coagulation time reagent.

Abstract: A reagent for determining prothrombin time (PT) including a recombinant protein tissue factor, a mixture of synthetic phospholipids, and a beta, gamma, or delta amino acid stabilizing compound is described for monitoring extrinsic blood coagulation activities. The source for the recombinant protein tissue factor is rabbit brain, and the phospholipids employed are palmitoyloleoylphosphatidylcholine (POPC) and palmitoyloleoylphosphatidylserine (POPS). The particular formulation buffer used to dilute the lipidated tissue factor provides a reagent that is optionally dried without lyophilization and remains stable for at least about 2 weeks at 37 C as either a liquid or a dried powder. A method for preparing the improved PT reagent and a method of using the reagent to analyze blood PT is also provided.

Abstract: A method for manufacturing a tissue factor-based prothrombin-time reagent includes combining predetermined quantities of tissue factor (TF), phospholipid and detergent-containing buffer to create a TF/phospholipid mixture. In this combining step, the quantity of TF is predetermined based upon its measured TF activity. Next, the detergent is removed from the TF/phospholipid mixture to produce an essentially detergent-free TF/phospholipid mixture that is useful as a tissue factor-based PT reagent. The method can include the use of hydrophobic resin that has been prescreened based on phospholipid recovery to accomplish the removal of the detergent.

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 producing a protein having an antithrombotic activity, which comprises replacing, in a protein that has an amino acid sequence having a homology of not less than 30% to the amino acid sequence of SEQ ID NO: 1 and forms a higher order structure composed of a first &bgr; strand (&bgr;1), a first &agr; helix (&agr;1), a second &agr; helix (&agr;2), a second &bgr; strand (&bgr;2), a loop, a third &bgr; strand (&bgr;3), a fourth &bgr; strand (&bgr;4) and a fifth &bgr; strand (&bgr;5) in this order from the amino terminus, at least one amino acid residue in a region from &agr;2 to &bgr;2 and/or a region from &bgr;3 to &bgr;4 so that electric charge of the amino acid residue is changed towards positive direction.

Abstract: A method is disclosed for determining if a patient is hypercoagulable, hypocoagulable or normal. The test involves providing a test sample from the patient and initiating coagulation in the sample in the presence of an activator, which is added to the sample in an amount which will result in intrinsic tenase-dependent fibrin. Then the formation of the intrinsic tenase-dependent fibrin polymerization is monitored over time so as to derive a time-dependent profile, with the results of the fibrin polymerization monitoring determining whether the patient is hypercoagulable, normal or hypocoagulable. The coagulation activator is added in an amount that triggers a thrombin explosion that is dependent on the propagation phase and amplification pathways. In this way, a single assay can assess the hemostatic potential of a sample.

Abstract: The present application relates to a method of making liposomes having membrane proteins incorporated therein, the method comprising: providing the membrane protein in solution; providing a solution of preformed liposomes; and incubating the mixture. Prior to the step of providing a solution of preformed liposomes, the liposomes are formed by combining a mixture of phospholipids with a solution of at least one type of unsaturated fatty acid. The methods of the present invention further relate to the method of making a reagent comprising tissue factor reconstituted into preformed liposomes. The method of the present invention for making a tissue factor reagent comprises: providing tissue factor in solution; providing a solution of preformed liposomes comprising a mixture of phospholipids and at least one type of unsaturated fatty acid; and incubating the mixture.

Abstract: A method for determining the acceptability of signal data collected from a prothrombin time (PT) test strip includes collecting PT signal data from an assay area and control areas of the PT test strip. Relationships between PT signal data collected from the assay area and PT signal data collected from each of the control areas (e.g., a ratio of post-peak pseudo-slopes) are then ascertained. A determination is subsequently made, based on the relationships, as to whether the PT signal data collected from the assay area is acceptable or not for use in calculating a PT and/or an International Normalization Ratio (INR). This determination can include, for example, comparing the relationships to at least one predetermined threshold limit. A prothrombin time (PT) monitoring system includes a PT test strip and a device. The PT test strip includes an assay area and a control area. The device (e.g., a portable handheld meter) includes an optical measurement means (e.g.

Abstract: A method of determining a measure of the number of platelets in a cell suspension containing platelets. The number of small particles in the suspension is counted. The suspension is agitated in the presence of a gas. The number of small particles in the suspension after agitation is counted. The two counts are compared to obtain a measure of the number of platelets.

Abstract: This invention provides a method for inhibiting thrombosis in a patient whose blood is subjected to extracorporeal blood circulation which comprises contacting the extracorporeal circulating blood with a Factor IXa compound in an amount effective to inhibit thrombosis in the patient. The Factor IXa compound may include an active site-blocked Factor IXa compound or Glu-Gly-Arg chloromethyl ketone-inactivated human factor IXa compound. This invention also provides that the effective amount may be from about 0.1 &mgr;g/ml plasma to about 250 &mgr;g/ml plasma or from about 0.5 &mgr;g/ml plasma to about 25 &mgr;g/ml plasma. The patient may be subjected to extracorporeal blood circulation during transplant surgery or cardiopulmonary bypass surgery or any surgery in which obligate clamping of a blood vessel is required. This invention further provides for a pharmaceutical composition which includes an effective amount of a Factor IXa compound and a pharmaceutically acceptable carrier.

Abstract: Platelet activation is measured by determining Mean Platelet Component (MPC) of suspended blood platelets, using a specific anticoagulant composition. The composition comprises at least one component for effecting platelet sphering (for example EDTA), and at least one platelet antagonist (for example at least one of, and preferably all three of theophylline, adenosine and dipyridamole).

Abstract: A prothrombin time reagent for determination of low molecular weight heparin in fresh whole blood and in anti-coagulant treated blood is provided. The reagent is composed of recombinant animal tissue factor, and a mixture of synthetic phospholipids, which mixture includes a phosphatidylalcohol. A formulation buffer which includes a sensitivity adjuster is used in formulating the reagent. The recombinant animal tissue factor includes rabbit brain. The synthetic phospholipids of the mixture include palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylserine (POPS), and a phosphatidylalcohol. The phosphatidyl alcohol includes dioleoylphosphatidylethanol, dioleoylphosphatidylmethanol, dioleoylphosphatidylpropanol, dioleoylphosphatidylbutanol, and dioleoylphosphatidylinositol. The sensitivity adjuster included in the formulation buffer is &ggr;-Cyclodextrin. The formulated reagent is air-dried and remains stable for at least 3 weeks at 37° C.

Abstract: The invention relates to novel recombinant glycoproteins that act as messenger substances, signaling substances, promoters, stimulators and initiators in a multitude of ways in the animal, especially human, circulation system.

Abstract: The invention relates to the use of monodisperse polymer particles of human albumin, fibrin, fibrinogen or polyphenylmethacrylate for quantitative measuring of the adhesion of platelets in vitro.

Abstract: The present invention is a container assembly that includes a coating composition on an interior surface of the container assembly and more particularly a coating composition for rapid clotting of a blood sample within a blood collection tube. The coating composition includes an intrinsic coagulation activator and an extrinsic coagulation activator for activating intrinsic and extrinsic pathways for promoting the rapid clotting of the blood sample. In addition, the present invention includes a barrier package for enclosing the container assembly.

Abstract: In connection with a fluidic medical diagnostic device that permits measurement of the coagulation time of blood, software, methods and associated devices for quality control are disclosed. The fluidic device preferably includes a test strip with one end having a sample port for introducing a sample and a bladder at the other end for drawing the sample to a measurement area. A channel carries sample from the sample port to an assay measurement area and first and second control measurement areas. Preferably, a stop junction, between the measurement areas and bladder, halts the sample flow for measurement. If results from measurements taken for each control fall within a predetermined zone or defined limits, the assay measurement is qualified. If not, an error is registered and the test strip is counted as unfit.

Abstract: A method, kit, system and reagent for measuring low molecular weight heparin in a whole blood sample is provided which involves the use of a Factor Xa activator, such as Russell's Viper Venom, as the coagulation assay initiator.

Abstract: In connection with a fluidic medical diagnostic device that permits measurement of the coagulation time of blood, software, methods and associated devices for quality control are disclosed. The fluidic device preferably includes a test strip with one end having a sample port for introducing a sample and a bladder at the other end for drawing the sample to a measurement area. A channel carries sample from the sample port to an assay measurement area and first and second control measurement areas. Preferably a stop junction, between the measurement areas and bladder, halts the sample flow for measurement. If results from measurements taken for each control fall within a predetermined zone or defined limits, the assay measurement is qualified. If not, an error is registered and the test strip is counted as unfit.

Abstract: Thrombotic or thromboembolic disease is detected or monitored by determining the presence or amount B in a urine sample.

Abstract: The present invention is directed to compositions and methods for measuring enzymatic activity, particularly glycosidase activity. Methods of the present invention include assays for quantitatively determining the amount of glycosidase activity in a sample. The present invention also provides methods for the diagnosis of metastatic and inflammatory processes in vitro and in vivo. The present invention further provides compositions and methods for high throughput assays for identifying compounds that effect glycosidase activity.

Abstract: A hemostasis analyzer, such as the Thrombelastograph® (TEG®) hemostasis analyzer is utilized to measure continuously in real time, the hemostasis process from the initial fibrin formation, through platelet-fibrin interaction and lysis to generate blood hemostasis parameters. The measured blood hemostasis parameters permit evaluation of platelet inhibition therapy.

Abstract: A platelet function assay reagent is provided for performing a platelet function assay, wherein the reagent contains a mixture of magnetic and non-magnetic particles, wherein the magnetic particles have bound to an outer surface thereof an amount of a first ligand having an affinity for direct interaction with GP-Ib receptors on blood platelets and wherein the non-magnetic particles have bound to an outer surface thereof an amount of a second ligand having an affinity for direct interaction with GP-Ib receptors on blood platelets, such that interaction of either of the first or second ligands with the GP-Ib platelet receptor will activate the blood platelets toward aggregation, wherein the first ligand and the second ligand can be the same or different, and the assay using such reagent, for providing a fast, reliable point-of-care assessment of platelet function.

Abstract: A reagent and kit are disclosed for determining if a patient is hypercoagulable, hypocoagulable or normal. The test involves providing a test sample from the patient and initiating coagulation in the sample in the presence of an activator, which is added to the sample in an amount which will result in intrinsic tenase-dependent fibrin. Then the formation of the intrinsic tenase-dependent fibrin polymerization is monitored over time so as to derive a time-dependent profile, with the results of the fibrin polymerization monitoring determining whether the patient is hypercoagulable, normal or hypocoagulable. The coagulation activator is added in an amount that triggers a thrombin explosion that is dependent on the propagation phase and amplification pathways. In this way, a single assay can assess the hemostatic potential of a sample.

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: Platelet contractile force (PCF) is used as a surrogate marker of thrombin generation. PCF generation occurs concomitant with the burst of prothrombin fragment F 1+2 release. The time between assay start and PCF onset is identified as the thrombin generation time (TGT), and is used in assessing risk of bleeding, in diagnosing various disorders, and in monitoring the effects of pharmaceutical and other treatments. TGT is prolonged in clotting factor deficiencies and in the presence of direct and indirect thrombin inhibitors. TGT shortens to normal with clotting factor replacement and shortens with administration of rVIIa. TGT is short in thrombophilic states such as coronary artery disease, diabetes and thromboangiitis obliterans and prolongs toward normal with oral and intravenous anticoagulants.

Abstract: Methods for monitoring the immune response and predicting clinical outcomes for patients on immunosuppressive drugs (such as transplant patients) are provided. The methods are based on the measurement of an intracellular metabolic marker in lymphocytes (such as ATP) as an indicator of a patient's immune response.

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: The present invention relates to procedures for the detection or for the determination of solid phase-associated factors, which are multiply associated with the same solid phase. According to the invention, the sample is brought into contact with a transmitter particle, on which at least one ligand having binding affinity for a solid phase-associated factor and a transmitter are immobilized, and a receiver particle, on which at least one ligand having binding affinity for said solid phase-associated factor and a receiver is immobilized, and then the signal is determined which results when transmitter and receiver are brought sufficiently close to one another. In particular, the invention relates to the detection of cell surface receptors which can be used for the typing of cells or for the determination of cell activation states. It is thus possible to replace the hitherto widely customary flow cytometry by a more simple procedure.

Abstract: The invention relates to a method for measuring the activity of the blood clotting factor XIIIa. The aim of the invention is to provide a method for measuring the activity of FXIIIa which is simple to carry out and which is also suitable for use with testing systems that have a high throughput. It was found that activated FXIIIa influences the light transmitting power of fibrin clots. Fibrinogen is caused to clot with a fibrinogen-splitting enzyme. The change in the light transmitting power of the samples according to the concentration of FXIIIa present is registered and is used as a measure of the activity of FXIIIa. The inventive method can be used especially for screening for FXIIIa inhibitors.

Abstract: Methods for controlling blood coagulation, and suitable pharmaceutical compositions that include a polypeptide that enhances the anticoagulation process (or inhibitors thereof for reversing the anticoagulation process) are provided.

Abstract: A system for performing a blood coagulation assay, having: a reaction chamber; at least one moveable member configured to mix contents of the reaction chamber; a sensor configured to detect the presence of a blood clot formed in the reaction chamber; and a timer that measures an interval of time between when a blood sample is received into the reaction chamber and when the sensor detects the blood clot formed in the reaction chamber.

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: The invention concerns contact activators for the endogenous coagulation pathway and their use in exploring coagulation anomalies.

Abstract: An analyzing method of a blood coagulation reaction by detecting an optical change of a blood sample with an elapse of time, the method comprises: setting at least one checkpoint or check region between a starting point of the blood coagulation reaction and the endpoint thereof; and monitoring a reaction state of the blood coagulation reaction at the checkpoint or in the check region to detect an abnormality of the blood coagulation reaction.

Abstract: A method of determining a measure of the number of platelets in a cell suspension containing platelets. The number of small particles in the suspension is counted. The suspension is agitated in the presence of a gas. The number of small particles in the suspension after agitation is counted. The two counts are compared to obtain a measure of the number of platelets.

Abstract: The present invention describes novel compositions and methods to enhance the in vitro and in vivo production of platelets and/or proplatelets from megakaryocytes. The present invention describes compositions comprising megakaryocytes, nitric oxide donors (i.e. compounds that donate, transfer or release nitric oxide, elevate endogenous levels of endothelium-derived relaxing factor, stimulate endogenous synthesis of nitric oxide or are substrates for nitric oxide synthase), and, optionally, at least one thrombopoiesis stimulating factor. The thrombopoiesis stimulating factor is preferably thrombopoietin. The nitric oxide donor is preferable S-nitrosoglutathione. The present invention also describes compositions comprising at least one nitric oxide donor and at least one thrombopoiesis stimulating factor. The present invention also provides methods for treating and/or preventing blood platelet disorders, and for producing platelets and/or proplatelets in vitro and in vivo.

Abstract: A method, kit, system and reagent for measuring low molecular weight heparin in a whole blood sample is provided which involves the use of a Factor Xa activator, such as Russell's Viper Venom, as the coagulation assay initiator.

Abstract: A method and device for testing a sample of fresh whole blood. In particular the present invention provides a method and device for testing a sample of fresh whole blood to determine whether a patient would benefit from the administration of a blood factor (such as AT III.

Abstract: The present invention relates to a treatment for myocardial infarction and blood clots within a patient, and more specifically to a therapy which enhances clot lysis comprising administering to a patient an antibody directed to &agr;2-antiplasmin crosslinked to fibrin (&agr;2AP-FX) which does not inhibit plasma &agr;2-antiplasmin (&agr;2AP). The invention also relates to a treatment for enhancing clot lysis comprising administering an antibody directed toward &agr;2-antiplasmin crosslinked to fibrin which does not inhibit plasma &agr;2AP together with a thrombolytic agent.