Two-Stage Procedure for the Quantitative Determination of Autoprothrombin III Concentration and Some Applications

1967 ◽  
Vol 18 (01/02) ◽  
pp. 198-210 ◽  
Author(s):  
Ronald S Reno ◽  
Walter H Seegers
Keyword(s):  
Prothrombin Time ◽  
Factor Vii ◽  
Two Stage ◽  
Pronounced Increase ◽  
One Stage ◽  
Assay Procedure ◽  
Bovine Plasma ◽  
The One ◽  
Autoprothrombin C

SummaryA two-stage assay procedure was developed for the determination of the autoprothrombin C titre which can be developed from prothrombin or autoprothrombin III containing solutions. The proenzyme is activated by Russell’s viper venom and the autoprothrombin C activity that appears is measured by its ability to shorten the partial thromboplastin time of bovine plasma.Using the assay, the autoprothrombin C titre was determined in the plasma of several species, as well as the percentage of it remaining in the serum from blood clotted in glass test tubes. Much autoprothrombin III remains in human serum. With sufficient thromboplastin it was completely utilized. Plasma from selected patients with coagulation disorders was assayed and only Stuart plasma was abnormal. In so-called factor VII, IX, and P.T.A. deficiency the autoprothrombin C titre and thrombin titre that could be developed was normal. In one case (prethrombin irregularity) practically no thrombin titre developed but the amount of autoprothrombin C which generated was in the normal range.Dogs were treated with Dicumarol and the autoprothrombin C titre that could be developed from their plasmas decreased until only traces could be detected. This coincided with a lowering of the thrombin titre that could be developed and a prolongation of the one-stage prothrombin time. While the Dicumarol was acting, the dogs were given an infusion of purified bovine prothrombin and the levels of autoprothrombin C, thrombin and one-stage prothrombin time were followed for several hours. The tests became normal immediately after the infusion and then went back to preinfusion levels over a period of 24 hrs.In other dogs the effect of Dicumarol was reversed by giving vitamin K1 intravenously. The effect of the vitamin was noticed as early as 20 min after administration.In response to vitamin K the most pronounced increase was with that portion of the prothrombin molecule which yields thrombin. The proportion of that protein with respect to the precursor of autoprothrombin C increased during the first hour and then started to go down and after 3 hrs was equal to the proportion normally found in plasma.

INHERITED FACTOR-VII DEFECT IN A NEGRO FAMILY

PEDIATRICS ◽  
1961 ◽  
Vol 27 (2) ◽  
pp. 204-213
Author(s):  
Helen I. Glueck ◽  
James M. Sutherland
Keyword(s):  
Prothrombin Time ◽  
Factor Vii ◽  
Homozygous State ◽  
Control Value ◽  
One Stage ◽  
First Case ◽  
Severe Deficiency ◽  
Low Levels ◽  
The Difference ◽  
The One

A case of factor-VII deficiency of a congenital nature in a Negro male child has been reported. As far as can be determined, this is the first case reported in this race. The defect was detected at 6 hours of age. Prothrombin, as contrasted to factor VII, after initially low levels normally found in infants, rose to adult levels. The patient's one-stage prothrombin time has ranged between 25 to 35 second (normal 11 to 12 seconds). In spite of this, he has never shown any manifestations of hemorrhage. The patient's family was studied and the findings indicate that the patient's defect represented a homozygous state and that both parents with a less severe deficiency were heterozygous for the trait. The defect is an autosomal disorder directly inherited. It is clinically apparent and easily detected only in the homozygous state. The heterozygous state is characterized by a very slight prolongation of the one-stage prothrombin time, the difference from the control value being so minimal as to be overlooked. In one subject studied, an aunt of the propositus, the quantitative defect (42% of normal) could not be regularly detected by the usual methods. Only by using the plasma of the propositus as the test plasma, was the defect in her plasma detected, thus explaining the transmission of the trait to her offspring. These findings explain the difficulties previously encountered in understanding the inheritance of the disorder.

Biological Properties of the Thromboplastins and Plasmas Included in the ICTH/ICSH Collaborative Study on Prothrombin Time Standardization

1979 ◽  
Vol 42 (04) ◽  
pp. 1115-1127 ◽  
Author(s):  
E A Loeliger ◽  
L P van Halem-Visser
Keyword(s):  
Prothrombin Time ◽  
Collaborative Study ◽  
Biological Properties ◽  
Bovine Brain ◽  
Factor Vii ◽  
Factor X ◽  
One Stage ◽  
Activation Products ◽  
Position Sensitivity ◽  
The One

SummaryThe fourteen types of thromboplastin included in the ICTH/ICSH Collaborative Study on Prothrombin Time Standardization were investigated with respect to their sensitivity for factor VII, activation products, and PIVKAs. All of these thromboplastins displayed sufficient factor VII sensitivity, and all were sensitive to activation products. After correction for the overall sensitivity slope, rabbit plain thromboplastins were the most sensitive preparations and human as well as rabbit diluted the least sensitive; bovine brain thromboplastin lost its exceptional position. Sensitivity to activation products explains why factors II, VII, and X present in artificially prepared abnormal plasmas may be difficult to assess accurately. Also, all thromboplastins appear to be sensitive for both PIVKA VII and PIVKA X. PIVKA VII acts as a procoagulant which increases the amount of factor VII measured by the one-stage assay technique. PIVKA X inhibits and thus reduces the activity of factor X. The procoagulant activity of PIVKA VII is particulary pronounced for the two lung-brain rabbit thromboplastins (Lyoplastin and Simplastin), whereas the anticoagulant action of PIVKA X is strongest with bovine brain thromboplastin (Thrombotest). Considered as a group, rabbit thromboplastins appear to be less sensitive for PIVKA X and more sensitive for PIVKA VII.As judged from thromboplastin calibration results, all three types of lyophilized reference plasmas are to some degree distinguishable from fresh patient plasmas. Primary calibration of a thromboplastin must therefore still be performed with freshly prepared normal as well as patient plasmas.

ACTIVATION OF FACTOR VII BY ASBESTOS IN BEEF PLASMA AND SERUM

1958 ◽  
Vol 36 (9) ◽  
pp. 953-958 ◽  
Author(s):  
L. G. Israels ◽  
E. Friesen ◽  
C. Sinclair
Keyword(s):  
Prothrombin Time ◽  
Human Plasma ◽  
Factor Vii ◽  
One Stage ◽  
Tissue Thromboplastin ◽  
The One

The adsorption of beef plasma on asbestos markedly shortens the one-stage prothrombin time of the plasma when tissue thromboplastin is used as the throm boplastic agent. This adsorption on asbestos does not affect the Russell viper venom time of the plasma. The adsorbed plasma exhibits increased factor VII activity over that of the original plasma. An eluate can be prepared from the asbestos which prolongs the one-stage prothrombin time sof beef and human plasma.

The Stuart-Prower Factor Assay and its Clinical Significance

1958 ◽  
Vol 02 (01/02) ◽  
pp. 024-038 ◽  
Author(s):  
F Bachmann ◽  
F Duckert ◽  
F Koller
Keyword(s):  
Prothrombin Time ◽  
Clinical Significance ◽  
Assay System ◽  
Factor Vii ◽  
Factor V ◽  
Optimal Conditions ◽  
High Dilution ◽  
One Stage ◽  
Factor Assay

SummaryA simple one-stage test for the determination of the Stuart-Prower factor is described. The method is a modification of the classical Factor VII complex assay, using a Russell’s viper venom (Stypven)-cephalin reagent instead of brain thromboplastin. The optimal conditions for the assay system have been systematically investigated. A high dilution of RVV (1 : 200 000) has given the best results. The system is not influenced by different concentrations of fibrinogen, prothrombin, Factor V, Factor VII and plasma thromboplastin precursors other than Stuart-Prower factor. Variations of lipoid and/or platelet contents of the plasma do not alter the results. Stuart-Prower factor is not activated by glass contact. The method has proved most valuable in the detection of heterozygous Stuart-Prower factor deficiencies. Comparison with a specific Stuart-Prower factor assay using patient’s plasma deficient in Stuart-Prower factor gave variations not greater than 15%. If determined simultaneously with the Quick one-stage ‘prothrombin time’, the test may provide a better control of patients under treatment with dicoumarol derivatives.

Studies Regarding the Effect of Foreign-Surface Contact on the One-Stage Prothrombin Time Determination

1966 ◽  
Vol 15 (03/04) ◽  
pp. 442-450 ◽  
Author(s):  
J. N Shanberge ◽  
T Matsuoka
Keyword(s):  
Factor Viii ◽  
Prothrombin Time ◽  
Hemophilia B ◽  
Factor Vii ◽  
One Stage ◽  
Surface Contact ◽  
Time Determination ◽  
Foreign Surface ◽  
The One

SummaryThe shortening of the one-stage prothrombin time of plasma brought about by increased foreign surface contact is dependent upon “activation” of factor VII. This activation of VII is the result of a series of activations involving factors XII, IX and probably XI as well. In this it is similar to the activation of factor VIII. Although the prothrombin time in an individual with hemophilia B or the Hageman trait is ‘‘normal”, no shortening occurs after increased foreign surface contact. Factor VII deficient plasma with normal levels of factors XII and IX is “activated” by “contact” in proportion to the level of VII present. No increase in activity of factors II, V, or X was observed after increased foreign surface contact.

ACTIVATION OF FACTOR VII BY ASBESTOS IN BEEF PLASMA AND SERUM

1958 ◽  
Vol 36 (1) ◽  
pp. 953-958
Author(s):  
L. G. Israels ◽  
E. Friesen ◽  
C. Sinclair
Keyword(s):  
Prothrombin Time ◽  
Human Plasma ◽  
Factor Vii ◽  
One Stage ◽  
Tissue Thromboplastin ◽  
The One

The adsorption of beef plasma on asbestos markedly shortens the one-stage prothrombin time of the plasma when tissue thromboplastin is used as the throm boplastic agent. This adsorption on asbestos does not affect the Russell viper venom time of the plasma. The adsorbed plasma exhibits increased factor VII activity over that of the original plasma. An eluate can be prepared from the asbestos which prolongs the one-stage prothrombin time sof beef and human plasma.

The Reaction of Thiol Compounds with the Vitamin-K Dependent Clotting Factors

1969 ◽  
Vol 21 (03) ◽  
pp. 573-579 ◽  
Author(s):  
P Fantl
Keyword(s):  
Prothrombin Time ◽  
Vitamin K ◽  
Anaerobic Conditions ◽  
Clotting Factors ◽  
Thiol Compounds ◽  
Aerobic Conditions ◽  
One Stage ◽  
Factor Ii ◽  
Ph Dependent ◽  
The One

SummaryTreatment of human and dog oxalated plasma with 0.2 to 1.0 × 10−1 M 2.3-dithiopropanol (BAL) or dithiothreitol (DTT) at 2–4° C for 30 min results in the reduction of the vitamin-K dependent clotting factors II, VII, IX and X to the respective-SH derivatives. The reaction is pH dependent. Under aerobic conditions the delayed one stage prothrombin time can be partly reversed. Under anaerobic conditions a gradual prolongation of the one stage prothrombin time occurs without reversal.In very diluted plasma treated with the dithiols, prothrombin can be converted into thrombin if serum as source of active factors VII and X is added. In contrast SH factors VII, IX and X are inactive in the specific tests. Reoxidation to active factors II, VII, IX and X takes place during adsorption and elution of the SH derivatives. The experiments have indicated that not only factor II but also factors VII, IX and X have active-S-S-centres.

Use of Different Tissue Thromboplastins in the Control of Anticoagulant-Therapy

1958 ◽  
Vol 02 (05/06) ◽  
pp. 462-480 ◽  
Author(s):  
Marc Verstraete ◽  
Patricia A. Clark ◽  
Irving S. Wright
Keyword(s):  
Prothrombin Time ◽  
Anticoagulant Therapy ◽  
Factor Vii ◽  
Rabbit Brain ◽  
Minor Role ◽  
Rabbit Lung ◽  
Coagulation Mechanism ◽  
Time Test ◽  
The One ◽  
A Minor

SummaryAn analysis of the results of prothrombin time tests with different types of thromboplastins sheds some light on the problem why the administration of coumarin is difficult to standardize in different centers. Our present ideas on the subject, based on experimental data may be summarized as follows.Several factors of the clotting mechanism are influenced by coumarin derivatives. The action of some of these factors is by-passed in the 1-stage prothrombin time test. The decrease of the prothrombin and factor VII levels may be evaluated in the 1-stage prothrombin time determination (Quick-test). The prolongation of the prothrombin times are, however, predominantly due to the decrease of factor VII activity, the prothrombin content remaining around 50 per cent of normal during an adequate anticoagulant therapy. It is unlikely that this degree of depression of prothrombin is of major significance in interfering with the coagulation mechanism in the protection against thromboembolism. It may, however, play a minor role, which has yet to be evaluated quantitatively. An exact evaluation of factor VII is, therefore, important for the guidance of anticoagulant therapy and the method of choice is the one which is most sensitive to changes in factor VII concentration. The 1-stage prothrombin time test with a rabbit lung thromboplastin seems the most suitable method because rabbit brain preparations exhibit a factor VII-like activity that is not present in rabbit lung preparations.

Procedures for the Quantitative Determination of Autoprothrombin C

1962 ◽  
Vol 08 (03) ◽  
pp. 434-441 ◽  
Author(s):  
Edmond R Cole ◽  
Ewa Marciniak ◽  
Walter H Seegers
Keyword(s):  
Quantitative Determination ◽  
Plasma Sample ◽  
Quantitative Measure ◽  
The Other ◽  
Clotting Time ◽  
Bovine Plasma ◽  
Autoprothrombin C

SummaryTwo quantitative procedures for autoprothrombin C are described. In one of these purified prothrombin is used as a substrate, and the activity of autoprothrombin C can be measured even if thrombin is in the preparation. In this procedure a reaction mixture is used wherein the thrombin titer which develops in 20 minutes is proportional to the autoprothrombin C in the reaction mixture. A unit is defined as the amount which will generate 70 units of thrombin in the standardized reaction mixture. In the other method thrombin interferes with the result, because a standard bovine plasma sample is recalcified and the clotting time is noted. Autoprothrombin C shortens the clotting time, and the extent of this is a quantitative measure of autoprothrombin C activity.

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