Thrombosis and Bleeding Disorders

Preface

Chapter I

Physiology and Biochemistry of Blood Coagulation

Introduction

Historical Aspects of Blood Coagulation

Nomenclature

Physicochemical Properties of Coagulation

Constituents

Activation of Prothrombin

Conclusions and Summary

Chapter II

Equipment and General Requirements of the Coagulation Laboratory

Introduction

Equipment

Collection of Blood

Cleaning and Preparation of Glassware

Buffer Solutions

Anticoagulants

Ion Exchange Resins

Chapter 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)

Discussion

Chapter 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)

Comments

Chapter 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 Inhibitors

Chapter VIII

Physiology and Biochemistry of Fibrinolysis

Introduction

General Scheme and Nomenclature

Individual Components

Summary and Conclusions

Fibrinolytic Activity in Whole Blood, Dilute Blood, and Ługlobulm 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-Products

Chapter 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 Fibrin

Chapter 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

Comments

Chapter 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 Proteins

Index