Abstract: The present invention features isolated angiogenesis inhibitors having a molecular weight of between about 40 kDa to 50 kDa and having an amino acid sequence substantially similar to that of the amino acid sequence shown in SEQ ID NO. 2 or SEQ ID NO. 3. Further provided are methods of making and using the angiogenesis inhibitors, e.g., to inhibit vascularization or to block osteonectin and plasminogen interaction.

Abstract: Methods for diagnosing and identifying genetic and metabolic factors associated with a physiologic procoagulant predisposition for and concurrent activation of the coagulation response in patients suffering from conditions such as chronic fatigue syndrome, fibromyalgia, Gulf War illness and cardiovascular disease are disclosed. Diagnostic assays utilized in the methods include measurement of blood levels of Protein C, Protein S, antithrombin, activated protein C resistance, prothrombin plasminogen activator inhibitor-1, lipoprotein (a) and homocysteine. Treatment regimens include anticoagulant therapies comprising administering warfarin or heparin as needed.

Abstract: An assay device and method are provided which allow the determination of the presence or absence of at least one analyte in a test sample, while providing specific identification of the test subject. The assay device includes a reaction medium having at least one reaction zone and at least one control zone, which is capable of providing a pattern suitable for identifying the test subject. The pattern suitable for identifying the test subject is preferably a fingerprint. In a preferred embodiment of the invention, the reaction zone and the control zone include at least one member of a ligand/receptor pair.

Abstract: Electrochemical system for measuring a value representing the coagulation time of a drop of whole blood including: an electrochemical sensor in the shape of a strip of small dimensions bearing at least a reference electrode and a working electrode on which a specific reagent is immobilized, including at least one chemical substrate one terminal link of which can be cut off by the thrombin enzyme to give charged groups (LG), and a measuring apparatus intended to receive said sensor allowing a given voltage or current to be imposed across the electrodes of the sensor, the electric signal resulting from the migration of the charged groups to be processed, said signal to be correlated with a value representing the coagulation time, and said value to be displayed on a display panel. The specific reagent preferably includes an oligopeptidic derivative as a substrate, a thromboplastin and a buffer medium.

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

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: 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: 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: 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: 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 with an 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 is associated to one end of the column. A valve/pump arrangement for filling at least one test tube with a carrier and at least part of the measurement sample is connected downstream of the column, 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: The invention is based upon the discovery that PAI-1 activity of a sample can be measured with sensitivity and correlation to in vivo activity without the use of standard curves. The assay determines the PAI-1 activity of a sample upon utilizing the second order rate equation for the reaction of PAI-1 and t-PA, as measured by their activities, in that sample.

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 relates to novel compounds and pharmaceutical composition, their preparation, and their use, having a antithrombotic effect through reversible inhibition of activated blood coagulation factor VIIa “FVIIa”.

Abstract: The present invention provides methods for diagnosing and/or monitoring thrombophilic disease in a patient that can result from the antiphospholipid antibody syndrome (aPL syndrome). The methods of the invention are premised on the inhibition of binding of an anticoagulant protein, annexin, preferably annexin-V, to phospholipids by antiphospholipid (aPL) antibodies in a patient blood sample.

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 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: The present invention relates to thrombin-containing hemostatic compositions, their preparation and use. In particular, it relates to hemostatic compositions comprising stabilized thrombin and microfibrillar collagen in an aqueous medium. In a preferred embodiment of the present invention, the compositions are used in a kit comprising two different components, one of which is autologous patient's plasma as the source of fibrinogen, and the other of which is the thrombin-containing composition which also contains microfibrillar collagen.

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: 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: 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: 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: For use in a lipid-dependent diagnostic assay, a stable aqueous suspension of a phospholipid which normally has a hexagonal (HII) organization when dispersed in an aqueous medium without detergent, the suspension containing the phospholipid, a detergent, and an aqueous phase. In the stable suspension, the phospholipid remains in suspension at a temperature of 25° C. for at least one hour. The suspension is suitable for providing the phospholipid to an assay for lupus anticoagulants which includes the step of pre-incubating a test sample with the phospholipid.

Abstract: A process for determining a resistance to activated protein C of a test specimen of human plasma following the steps of: (1) mixing together (a) the test specimen of human plasma, (b) a reactant deficient in factor V which supplies at least most of the coagulation factors other than factor V, and (c) the venom of Crotalus viridis helleri which specifically activates factor X to Xa, and incubating the mixture of (a), (b) and (c) for at least one minute at a temperature of between 10 and 45° C.; (2) introducing into the incubated mixture(i) Ca2+ or (ii) Ca2++exogenic activated protein C; and (3) determining the coagulation time (i) in the absence of activated protein C and (ii) in the presence of activated protein C. Steps (1) to (3) are repeated, but replacing, in step (1), the test specimen with a normal plasma as control and correlating resistance to activated protein C by comparing the determinations made in steps for the test specimen and for the normal plasma.

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: Persons having the Factor VLEIDEN mutation and a lower than average plasma level of TFPI have been found to be at elevated risk of thrombosis. Prophylactic methods for reducing the risk of thrombosis employ the administration of TFPI to achieve higher levels in the plasma.

Abstract: A cocktail reagent preparation for the rapid production of serum contains thrombin, snake venom, and protamine sulfate. The preparation employs very small quantities of clot promoting substances which behave in a synergistic manner such that rapid clotting of highly heparinized blood is achieved without altering the chemical analysis of the blood enzymes, proteins, sugars, or electrolytes. Thus, clinicians who rely upon the results of such tests can more closely monitor organ and tissue function and adjust patient therapies accordingly.

Abstract: An assay for compounds useful in the treatment of a bacterial induced coagulation disorder has the following steps: a) incubating a plasma sample with a strain of bacteria; b) adding a compound to be assayed to the plasma sample before, during or after step (a); c) conducting an activated partial thromboplastin time test; d) determining the clotting time.

Abstract: Methods for detecting and/or quantifying catalytically-active, serine proteases in a biological fluid are disclosed. The methods are useful for measuring the active enzymes of the coagulation/fibrinolytic system and evaluating the system or components of the system as indicative of thrombosis-related disorders. The methods involve the combined use of halomethyl ketone probes having broad specificity for catalytically-active serine proteases and immunological reagents specific for serine proteases of particular types. The halomethyl ketone probes are active site specific; they are only incorporated into catalytically-active serine proteases. An antibody is used to provide specificity for the particular type of serine protease. By the combined active-site-specificity of the halomethyl ketone probes and the type-specificity of the antibody, the catalytically-active fraction of a particular serine protease is determined.

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: A method for determining whether a subject has had a stroke and, if so, the type of stroke which includes analyzing the subject's body fluid for at least four selected markers of stroke, namely, myelin basic protein, S100 protein, neuronal specific enolase and a brain endothelial membrane protein such as thrombomodulin or a similar molecule. The data obtained from the analyses provide information as to the type of stroke, the onset of occurrence and the extent of brain damage and allow a physician to determine quickly the type of treatment required by the subject.

Abstract: An improved apparatus and method for evaluating platelet functionality of a blood sample. The apparatus includes a plurality of test cells. Each of the cells includes a platelet function restoration agent, an anticoagulant agent, and a clotting reagent. At least one of the cells also includes a platelet activating agent. The clotting time is determined for each of the aliquot portions, and the relative clotting times of the aliquot portions in the cells are determinative of the platelet functionality of the sample. The method includes the steps of combining a platelet function restoration agent, an anticoagulant agent, a platelet activating agent, and the sample of blood to be tested to form a test mixture. The platelets of the sample are activated by adding a clotting reagent to the test mixture at the start of the activated clotting time test, and the activated clotting time test is terminated upon detecting a predetermined change in a property of the test mixture.

Abstract: A method of determining a dose response for a platelet inhibitor. The method includes the steps of placing a predetermined amount of heparin in each cell of a multicell test cartridge, placing an optimized amount of a clotting activator in each cell, and placing a measured amount of platelet inhibitor in each cell, the amount of inhibitor 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 inhibitor 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 calculate and determine the platelet inhibition effect of the platelet inhibitor.

Abstract: A method for determining the endogenous thrombin potential of a sample having a total anticoagulant activity of or equivalent to at least 0.07 U ISH/ml, includes using a thrombin substrate or a salt thereof that is soluble in the sample to determine the ETP of the sample. Suitable thrombin substrate include those of the formula P-Val-Xaa-S, in which P is an amino protective group, that is non-aromatic and polar, Val is a valine residue attached via a peptide bond to Xaa, Xaa is an amino acid residue comprising a terminal guanidino group or ureido group separated by at least 2 carbon atoms from the peptide backbond the amino acid residue is attached to S and S is a signal group such as a chromophore that can be enzymatically hydrolyzed.

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 process for the production of plasmas with added annexins for use as a control or standard in all functional clotting tests which are used for the detection of a lupus anticoagulant.

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: The present invention relates to stable compositions useful as primary standards and calibrators and controls comprising a cardiac troponin I (cTnI) such as native, recombinant, addition and deletion forms thereof, whether or not complexed with other troponin subunits such as TnC and/or TnT, in an inactivated human serum. The compositions are obtained by incubating troponin complexes with human serum. The compositions are characterized by an immunodetectability ratio of epitopes on the N-terminal segment to epitopes on the C-terminal segment substantially equivalent to that of pooled, fresh serum from acute myocardial infarction patients.

Abstract: An additive formulation comprising heparinase and trehalose, a method for using the formulation and a device containing the formulation. The additive formulation is useful in substantially neutralizing residual heparin from a blood sample when used in a blood collection tube without interfering with the clinical analysis of the blood sample.

Abstract: The present invention relates to a method for identification of defects in the clotting, fibrinolysis and complement system.

Abstract: A method for preparation of an air-dried prothrombin time (PT) reagent which uses a recombinant protein and synthetic phospholipids is described. The source for the recombinant protein 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 dried without lyophilization and remains stable for at least 2 weeks at 37 C. A method for preparing the improved PT reagent and a method of using the reagent to analyze blood PT is also provided.

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 with an 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 is associated to one end of the column. A valve/pump arrangement for filling at least one test tube with a carrier and at least part of the measurement sample is connected downstream of the column, 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: The presence of endotoxin in the outer membrane of cell wall of a Gram-negative bacteria is used to determine bacterial contamination in a biological product. The amount of endotoxin present in a biological product is accurately measured without influence of a limulus reaction-activating substance which causes a false-positive reaction, by a method including the steps of: inactivating the limulus reaction-activating substance, if any, in the biological product, by exposing the biological product to a surfactant at a temperature ranging from the surfactant's freezing point to 50° C.; and measuring the amount of endotoxin present in the biological product, using a limulus reagent.

Abstract: A method of performing a fibrinogen assay is provided using an assay reagent containing ecarin to give an assay that is insensitive to the presence of heparin and insensitive to hematocrit, while being useful as a point of care assay in both a dry chemistry and wet chemistry format.

Abstract: An agent for dehydrating corneas, in particular eye corneas for organ culture, contains the culture medium and as dehydrating substance hydroxyethylstarch with a mean molecular weight Mw from 070,000 to 200,000, an MS substitution degree from 0.15 to 0.5, a DS substitution degree from 0.15 to 0.5 and a C2-C6 substitution ratio at the anhydroglucose units .ltoreq.8. The dehydrating substance is preferably hydroxyethylstarch with a mean molecular weight Mw 130,000.+-.20,000, an MS substitution degree from 0.38 to 0.45, a DS substitution degree from 0.32 to 0.40 and a C2-C6 substitution ratio at the anhydroglucose units from 8 to 20. The hydroxyethylstarch is used in a concentration from 1 to 20 (weight/vol.) %, preferably from 2 to 15 (weight/vol.) %, and in particular of 7.5% (weight/vol.).

Abstract: This invention provides a method to identify compounds potentially useful for the treatment of neoplasia in mammals. The phosphodiesterase inhibitory activity of a compound is determined along with COX inhibitory activity. Growth inhibitory and apoptosis inducing effects on cultured tumor cells are also determined. Compounds that exhibit phosphodiesterase inhibiton, growth inhibition and apoptosis induction, but not substantial prostaglandin inhibitory activity, are desirable for the treatment of neoplasia.

Abstract: This invention relates to a method of analyzing coagulative, fibrinolytic or haemostatic activity in, especially, blood or plasma from mammals, particularly humans. The method comprises bringing a sample, in vitro, into contact with fixed and preferably from their growth support detached endothelial cells or outer membranes of such cells, and detecting the resulting coagulated material and, in some cases, lyzed coagulated material. Reagents are also described that are based on the endothelial cells and their outer membranes. A kit for performing such a method including the claimed reagent is also described.

Abstract: A method for determining the amount of an antiplatelet compound in a subject being treated with the compound which comprises calculating the ACT number of the subject's blood containing a reagent which immunoreacts with the antiplatelet compound and comparing the figure to a standardized concentration figure. In subjects concurrently being treated with heparin and antiplatelet compounds, the method can be practiced without additional heparin added to the blood sample. The invention also provides reagents which immunoreact with antiplatelet compounds and kits comprising immunoreactive reagents.

Abstract: In a method of determining heparin content, a known amount of thrombin (FIIa) or activated coagulation factor X (FXa) is added to a mixture which comprises a known concentration of chromogenic substrate (S), a known concentration of antithrombin (AT) and a sample having an unknown heparin concentration. The amount of FIIa or FXa is chosen such that at most 20%, of the S present is reacted during the period which the AT needs to deactivate all the FIIa or FXa present. The final concentration of the reacted chromogenic substrate p-nitroanilide ([pNA].sub.final) is determined after completion of the reactions, and the [pNA].sub.final is used to determine the rate constant (k.sub.dec) of the reaction of FIIa or FXa with AT using the relationship: ##EQU1## The heparin concentration in the sample can then be determined using the value of k.sub.dec from a predetermined calibration curve of k.sub.dec against heparin concentration.

Abstract: An artificial liver support system is described herein which comprises cryopreserved hepatocytes having an initial viability of 80-99%. Further disclosed are hepatocytes cryopreserved by dispensing hepatocytes into freezing containers, freezing the containers from between minus 50 to minus 90 degrees Celsius, storing the containers in liquid or vapor nitrogen, thawing the cryopreserved hepatocytes when ready for use and removing residual cryoprotectant media.