Thrombosis and Bleeding Disorders

Thrombosis and Bleeding Disorders compiles the laboratory and research aspects of thrombosis and hemorrhagic disorders in humans. This book presents reviews of the underlying theory, physiology, and biochemistry of hemostasis and thrombosis, including the enzymology of blood coagulation and fibrinolysis. This compilation is divided into three levels of specific purposes. First is to provide the most reliable and widely accepted laboratory assays of undisputed diagnostic clinical value, which provides newcomers in the field and experienced workers in the coagulation laboratory with a reference manual to everyday work in a clinically-oriented environment. Second is to review and sketch in outline the theoretical sections focusing on mechanisms. Finally, this text aims to include a systematic review of the most successful purification techniques for individual coagulation factors and moieties of the fibrinolytic enzyme system. This publication is beneficial to medical students and clinicians concerned with human blood coagulation.

PrefaceChapter I Physiology and Biochemistry of Blood Coagulation Introduction Historical Aspects of Blood Coagulation Nomenclature Physicochemical Properties of Coagulation Constituents Activation of Prothrombin Conclusions and SummaryChapter II Equipment and General Requirements of the Coagulation Laboratory Introduction Equipment Collection of Blood Cleaning and Preparation of Glassware Buffer Solutions Anticoagulants Ion Exchange ResinsChapter III Clotting Time Techniques Introduction Whole Blood Coagulation Time Recalcification Time Heparin Sensitized Clotting Time Introduction Description of Methods General Discussion Thrombelastography Introduction Basic Description of Apparatus and Principle Techniques of Measurement Evaluation of the Thrombelastogram Abnormal Thrombelastographic Partial Thromboplastin Time Techniques Introduction Reagents Steps in Procedure Interpretation of Results Modifications Precautions Prothrombin Consumpdon Tests Introduction Measurement of Prothrombin Consumption Thrombin and Thromboplastin Generation Techniques Introduction Thrombin Generation Test Thromboplastin Generation Test Thromboplastin "Screening" Test (Thrombin- Thromboplastin Generation Test) Thromboplastin Activation Test One-Stage Prothrombin Time Techniques Introduction Practical Applications The Unmodified One-Stage Test The "P and P" Method (Owren and Aas, 1951) The Thrombotest Method (Owren 1959)Chapter IV Purification of Prothrombin Introduction Methods of Purification) Discussion Purification of Prethrombin "Modified Zymogen" Introduction Methods of Purification Discussion Purification of Thrombin Introduction Methods of Purification Discussion Purification of AC-Globulin (Factor V) Introduction Method of Purification Discussion Purification of Factor VIII Introduction Discussion Purification of Factor VII and Factor IX Introduction Factor VII - Methods of Purification Factor IX-Methods of Purification Discussion (Autoprothrombin III-C) Purification of Factor X Introduction Methods of Preparation Discussion Purification of Hageman Factor (Factor XII ) Introduction Methods of Purification Purification of Plasma Thromboplastin Antecedent (PTA, Factor XI) Introduction Methods of Purification Discussion Purification of Tissue Thromboplastin Introduction Methods of Preparation Purification of Platelet Factor 3 Introduction Method of Purification (Alkjaersig et al 1955) DiscussionChapter V Assay for Prothrombin Introduction Original Two-Stage Procedure Other Two-Stage Assays Role of Autoprothrombin C in the Assays Thrombin Clotting Assays Introduction Methods Assays of Factor V Introduction Methods of Assay Assays for Antihemophilic Factor (Factor VIII) Partial Thromboplastin Method (Langdell et al. 1953) Thromboplastin Generation Method Addendum Partial Thromboplastin Method with Artificial Factor VIII - Free Reagents (Stapp, 1961a) Principle Assays for Factors VII and X Introduction Assay for Factor X Assays of Factor VII Assays for Factor IX (Christmas Factor, Plasma Thromboplastin Component [PTC], Platelet Cofactor II, Antihemophilic Globulin B [AHF B/J]) Introduction Assay for Factor IX by the Thromboplastin Generation Test (Biggs and Douglas, 1953) (modified by Schwick, 1954; and Schwick and Störiko, 1966) Assay for Factor IX by the Prothrombin Consumption Test Principle (Quick, 1947, 1966) Assays for Factor IX by the Partial Thromboplastin Time Assays for Factor IX using Artificial Substrates Assays for Autoprothrombin I, II, III, C, and Prethrombin Introduction Assay for Autoprothrombin Ip Assay for Autoprothrombin Ic Assay for Autoprothrombin II Assay for Autoprothrombin III Assay for Autoprothrombin C Assay for Prethrombin Assays for Hageman Factor (Factor XII) and Plasma Thromboplastin Antecedent (Factor XI) Introduction Assay for Hageman Factor (Adapted from Ratnoff, 1964) Assay for PTA (Adapted from Rapaport et al. 1961a)Chapter VI Fibrinogen Introduction Biophysical and Chemical Properties of Fibrinogen Fibrinogen-Fibrin Conversion Isolation and Assay of Fibrinogen Fibrinogen and Fibrin Derivatives Introduction Chemical Procedures Immunological Determination (Based on the Method of Schultze and Schwick, 1959) Presumptive Tests Isolation and Purification of Fibrin Stabdizing Enzyme (F. XIII, FSF, Laki-Lorand Factor, Fibrinase) Introduction Isolation of FSF (F. XIII) Assays for Fibrin Stabilizing Factor (Factor XIII) Introduction Substrates for the FSF Assay Assay Using FSF Free Fibrinogen (Loewy etal. 1961) Assay Using Plasma from Patients with FSF Deficiency Immuno Assay (Bohn and Haupt, 1968) CommentsChapter VII Determination of Antithrombin Introduction Methods of Measurement Heparin Assays in Blood Introduction Assays for Anticoagulant Activity of Heparin Control of Heparinemia Technical Procedures Chrculating Anticoagulants Introduction Demonstration of Inhibitors by Means of Test Systems Measuring the Overall Coagulability of Plasma Determination of the Site of Action of an Inhibitor Methods for the Biochemical Characterization of InhibitorsChapter VIII Physiology and Biochemistry of Fibrinolysis Introduction General Scheme and Nomenclature Individual Components Summary and Conclusions Fibrinolytic Activity in Whole Blood, Dilute Blood, and Luglobulm Clot Lysis Time Tests Introduction Blood Clot Lysis Time Diluted Blood Clot Lysis Time (Fearnley et al. 1957) Euglobulin Clot Lysis Time Discussion The Fibrin Plate Method for Assay of Fibrinolytic Agents Principle Reagents Preparation of Fibrinogen Technique of Assay Gomments The Puritication of Profibrinolysin and Fibrinolysin Introduction Physicochemical Characteristics Methods for the Purification of Profibrinolysin Purification of Fibrinolysin Evaluation of the Profibrinolysin Reagents Conclusions Gaseinolytic Techniques Introduction Plasminogen Plasmin Plasminogen Activators Plasminogen Assays Using Fibrin as a Substrate I ntroduction Method of Johnson and Tillet (1952); (Johnson and Tse, 1967) Method of Brakman Method of Celander and Guest (1959); (Guest, 1954) The Esterase Assays of Enzymes of Blood Clottmg and Lysis Glossary Introduction Experimental Methods for Assaying Esterase Action Methods Depending on the Alcohol Liberated Radioactive Methanol Method Discussion Assays of the Plasminogen Activator in Tissues Principles Reagents and Materials Preparation of Fibrin Plates Isolation of Tissue Activator Assay on Fibrin Plates The Clot Lysis Time Method Assay Methods for Individual Fibrinolytic Components - Urokinase and Streptokinase Introduction Urokinase Assay Streptokinase Assay Assay for Plasminogen Activators with Labeled Fibrin Substrates Introduction Radioiodine-Iabelled Fibrin Fluorochrome Labelled Fibrin Summary and Conclusions Streptokinase Tolerance Test Introduction Predicted Dose Test (Fletcher et al. 1958) (Modification Deutsch and Fischer, 1960) Streptokinase Tolerance Test using the Thrombelastograph (TEG) (Fischbacher 1960) Comment Determination of Inhibitors of Fibrinolysis Introduction Methods of Determination Differentiation Between Intravascular Coagulation and Intravascular Proteolysis Introduction Methods for Differentiation Techniques for Assay of Split-ProductsChapter IX Hemostasis Introduction Phylogenetic Aspects of Hemostasis Platelet Adhesion - the Interaction of Platelets with Surfaces other than those of other Platelets Platelet Aggregation Consolidation of the Platelet Plug Morphological Aspects of Hemostasis Summary and Conclusion Bleeding Time Techniques Introduction Methods Comments Tests for Capillary Frs^ility and Resistance Introduction The Pressure Method (Torniquet Test, Rumpel-Leede, Gothlin, Hess) The Suction Method Comments Platelet Count Techniques, Platelet Adhesiveness and Aggregation Tests Introduction Platelet Count Techniques Platelet Adhesiveness Tests Chandler's Tube Technique (1958) Quantitative Platelet Aggregation Tests Platelet Electrophoresis General Conclusions Assays for Platelet Factors Introduction Preparation of Platelet Suspensions Platelet Factor 1 Platelet Factor 2 Platelet Factor 3 Platelet Factor 4 (Deutsch, 1959) Platelet Fibrinogen Fibrm Stabilizing Factor Clot Retraction Introduction Methods Comments Electron Microscopic Techniques for Blood Platelets, Fibrinogen and Fibrin Introduction Isolation of Blood Platelets Fixation Dehydration and Embedding Staining Other Techniques in the Study of Blood Platelets Histochemical Studies Fibrinogen and FibrinChapter X Immunologic Techniques Introduction Antibody Production Antibody Characterization Immunochemical Tests - Qualitative Immunochemical Tests - Quantitative Fluorescent Antibody Technique Conclusion Immune Assays of Tissue Thromboplastin Introduction Hemagglutination Inhibition Reaction CommentsChapter XI Thrombosis Introduction Definition Pathology of Human Thrombi Experimental Thrombosis Etiology of Thrombosis Summary Methods for the Experimental Study of Intravascular Thrombus Formation Introduction Categories of Methods to Produce Experimental Thrombosis and Specific Examples Perivascular Applications of Thrombosing Substances Experimental Animal Models for the Production of Disseminated Intravascular Coagulation Introduction Pathophysiology Experimental Approach to DIC Localized Shwartzman Phenomenon Demonstration of Plasma Protein in Microscopic Sections with Emphasis on the Identification of Fibrin Introduction Demonstration of Fibrin Based on its Ultrastructure Demonstration of Fibrin by Specific - Histologic Stains Demonstration of Fibrin and other Proteins by Their Amino Acid Composition Demonstration of Specific Proteins as Substrates for Specific Proteases Immunochemical Demonstration of Specific ProteinsIndex