THE MOLECULAR WEIGHT DEPENDENT EFFECTS OF HEPARIN ON THROMBOPLASTIN ACTIVATED PROTEASES

1987 ◽  
Author(s):  
R M Emanuele ◽  
J C Lormeau ◽  
J Choay ◽  
J Fareed
Keyword(s):  
Molecular Weight ◽  
Prothrombin Time ◽  
Antithrombin Iii ◽  
Gel Filtration ◽  
Factor Vii ◽  
Fibrinopeptide A ◽  
Factor X ◽  
Dependent Effect ◽  
Generation Test ◽  
Different Molecular Weight

Five different molecular weight (M. W.) fractions of heparin (M. W.'s 23,000 ; 17,450 ; 13,300 ; 9,000 and 5,100) were prepared by gel-filtration. Screening of these fractions in global and amidolytic assaysat a concentration of 2.5 ug/ml revealed a dependence on M. W. for the potency response. In the APTT, TT, Heptest, amidolytic anti Xa and Ila assays, an increase in potency was observed with increasing M. W.. This relationship remained consistent to 13,300 M. W. after which no further increases in potency were observed. However, when the same fractions were screenedfor their effect on prothrombin time (PT), a different M. W. dependent pattern was observed. In this assay, a continued increase in potency was observed for the 17,450 and 23,000 M. W. fractions. At a concentration of 20 ug/ml, the 23,000 M. M. fraction produced the largest prolongation of the PT while the 5,100 M. W. fraction showed the least effect. The correlation between M. W. and potency in the PT assay was 0.98. No M. W. dependent potency response was observed whenidentical PT.'s were performed in plasma deficient in antithrombin-III. Further testing of the M. W. fractions using a Ca++/thromboplastin activated fibrinopeptide-A generation test resulted in a potency response similar to that observed in the PT. To more precisely investigate this M. W. dependent effect upon thromboplastin activation an amidolytic assay using factor VII-thromboplastin to convert factor X to Xa wasdeveloped. The inhibition of Xa in the presence of the M. W. fractions and antithrombin III was used as the assay endpoint. As with previous assays involvingthromboplastin activation, potency increased with increasing M. W. from 5,100 through 23,000. These observations coupled with the failure of amidolytic anti Xa assays to show increased potency beyond 13,000 M. W., suggest that heparin of high M. W. may beacting at the level of thromboplastin or factor VII / VIIa.

Hemostatic profile of bovine ovarian follicular fluid

1995 ◽  
Vol 73 (5) ◽  
pp. 624-629 ◽  
Author(s):  
Manabu Yamada ◽  
Patricia A. Gentry
Keyword(s):  
Prothrombin Time ◽  
Follicular Fluid ◽  
Inhibitory Activity ◽  
Antithrombin Iii ◽  
Factor Vii ◽  
Coagulation System ◽  
Factor X ◽  
Low Levels ◽  
Coagulation Pathway ◽  
Ovarian Follicular Fluid

The hemostatic profile of bovine ovarian follicular fluid was evaluated and the levels of procoagulant, fibrinolytic, and inhibitory activity compared with plasma. The results of the prothrombin time assay and the presence of fibrinogen along with factor VII and factor X activity indicate that bovine follicular fluid possesses components of the "extrinsic" or "tissue factor" coagulation system. The absence of factor VIII:C activity, along with the extremely low levels of factors IX and XI, indicates that there is not a functional "intrinsic" coagulation pathway. The fluid derived from large follicles exhibited increased levels of factors VII and X activity and a shorter prothrombin time compared with fluid obtained from the less mature small follicles. Similar alterations in the levels of the inhibitory proteins antithrombin III and α2-macroglobulin were observed. Overall the amount of antithrombin III was similar to that in plasma, the levels of fibrinogen and factor X were approximately 2-fold lower, and the levels of factor VII and factor X were approximately 10-fold lower than in plasma. The fibrinolytic activity in follicular fluid was greater than the procoagulant or inhibitory activity. Plasminogen activator activity was 5-foid higher, while both plasminogen and antiplasmin values were similar to plasma levels.Key words: hemostasis, follicular fluid, bovine.

Haemostatic Parameters and Lifestyle Factors in Elderly Men in Italy and The Netherlands

1996 ◽  
Vol 76 (03) ◽  
pp. 411-416 ◽  
Author(s):  
Fransje C H Bijnen ◽  
Edith J M Feskens ◽  
Simona Giampaoli ◽  
Alessandro Menotti ◽  
Flaminio Fidanza ◽  
...  
Keyword(s):  
Alcohol Use ◽  
The Netherlands ◽  
Antithrombin Iii ◽  
Lifestyle Factors ◽  
Activated Partial Thromboplastin Time ◽  
Factor Vii ◽  
Fat Intake ◽  
Factor X ◽  
Elderly Men ◽  
History Of

SummaryThe association between plasma fibrinogen, factor VII, factor X, activated partial thromboplastin time, antithrombin III and the lifestyle factors cigarette smoking, alcohol use, fat intake and physical activity was assessed in 802 men aged 70-90 years in Zutphen (The Netherlands), Montegiorgio and Crevalcore (Italy).Smoking was positively associated with fibrinogen, also after adjustment for other lifestyle factors, age, use of anticoagulants and aspirin like drugs, body mass index, and history of myocardial infarction. Alcohol use was associated with increased levels of factor X and decreased levels of antithrombin III. Fat intake was positively associated with antithrombin III. Between cohorts, considerable differences were observed in levels of haemostatic parameters and the lifestyle factors. Compared to the mediterranean cohorts the Zutphen cohort showed the highest levels of fibrinogen and factor VII. Differences in lifestyle factors could, however, not explain differences between cohorts in levels of any of the haemostatic parameters, despite the observed associations between lifestyle factors and haemostatic parameters.

The Different Forms of Antithrombin III in Serum

1977 ◽  
Vol 38 (02) ◽  
pp. 0494-0503 ◽  
Author(s):  
D. S Pepper ◽  
D Banhegyi ◽  
J. D Cash
Keyword(s):  
Molecular Weight ◽  
Affinity Chromatography ◽  
Human Serum ◽  
Degradation Product ◽  
Antithrombin Iii ◽  
Gel Filtration ◽  
Free Form ◽  
Polymeric Form ◽  
At Iii ◽  
Human Thrombin

SummaryAntithrombin III (AT III) complexes were isolated from human serum by affinity chromatography and gel filtration. In the first step of the preparation, using heparin-agarose chromatography, we observed that the complexed form of AT III bound less strongly to the gel than the free form and that about half of the AT III was free. With further purification a 2.5 × 105 molecular weight complex was isolated. Using 125I labelled human thrombin, this complex was radioactive indicating the presence of thrombin. Only in a synthetic thrombin-AT III system was a 9 × 104 molecular weight complex detected, but not in serum. These facts suggest that in serum AT III complexes may exist in a polymeric form. Also, an AT III antigen derived from the original AT III molecule, but not complexed, was isolated which may be a degradation product.Abbreviations used: AT-III, antithrombin III. Hepes, N-2-Hydroxyethylpiperazine-N-2-Ethanesulphonic acid.

scholarly journals The Heparin Inhibiting Property of Brain Thromboplastin

1977 ◽  
Author(s):  
E.D. Gomperts ◽  
M. Zucker
Keyword(s):  
Prothrombin Time ◽  
Antithrombin Iii ◽  
Proteinase Inhibitors ◽  
Serine Proteinase ◽  
Weight Fraction ◽  
High Molecular Weight Fraction ◽  
Thrombin Time ◽  
Factor X ◽  
Heparin Resistance

Antithrombin III is one of the serine proteinase inhibitors of the plasma which has been shown to specifically inhibit thrombin as well as Factor X. Heparin acts via antithrombin III, the heparin cofactor, hence it is difficult to explain the relative insensitivity of the prothrombin time to the presence of heparin in plasma as both thrombin, ana Factox Xa are associated functionally with the prothrombin time. This insensitivity becomes more obvious on appreciating the extreme sensitivity to heparin of the activated partial thromboplastin time as well as the thrombin time. This communication reports the demonstration of heparin inhibiting action of brain thromboplastin. The response of the prothrombin time to heparin under various conditions, and the effect of brain thromboplastin obtained from various sources and by different preparative techniques on the action of heparin in vitvo have been studied. The heparin inhibiting activity was shown to parallel the tissue factor activity. It is heat labile, non-dialysable, destroyed by detergent activity and lies in a high molecular weight fraction of the brain thromboplastin preparation (>300,000). In addition to explaining certain in vitro phenomena, these observations may explain the previously observed heparin resistance in the generalised Schwartzman phenomenon.

Effect of a Moderate Fish Intake on Haemostatic Parameters in Healthy Males

1989 ◽  
Vol 61 (03) ◽  
pp. 468-473 ◽  
Author(s):  
A D Muller ◽  
A C v Houwelingen ◽  
M C E van Dam-Mieras ◽  
B M Bas ◽  
G Hornstra
Keyword(s):  
Intervention Study ◽  
Dietary Intervention ◽  
Antithrombin Iii ◽  
Experimental Period ◽  
Procoagulant Activity ◽  
Factor Vii ◽  
Factor X ◽  
Activated Factor Vii ◽  
Von Willebrand ◽  
Willebrand Factor

SummaryThis article describes the results of a dietary intervention study performed in three different centers. In the study the effect of a diet enriched with fish on the coagulation tendency of blood was investigated. Two groups of 40 volunteers were given a dietary supplement consisting of 135 g of canned mackerel or meat paste (control) for a 6 weeks period.Compliance, monitored by measuring the urinary excretion of lithium, added to the supplements, was about 80%. Before, during and at the end of the experimental period a number of hemostatic parameters, reflecting the coagulation tendency of blood and the procoagulant activity of monocytes, were measured.The fish supplement did not cause a significant effect on the prothrombin time and on the levels of factor VII, activated factor VII, antithrombin III, von Willebrand factor, fibrinogen, plasminogen and a2-antiplasmin. A slight but transient prolongation in the activated partial thromboplastin time was observed as well as a significant increase in the factor X level, which became more pronounced with prolongation of the experimental period; no activated factor X was found.A tendency towards a stimulation of monocyte procoagulant activity was noticed.

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.

High Molecular Weight Forms of Antithrombin III Complexes in Blood

1983 ◽  
Vol 49 (01) ◽  
pp. 032-036 ◽  
Author(s):  
E Marciniak ◽  
G Gora-Maslak
Keyword(s):  
Antithrombin Iii ◽  
Serum Proteins ◽  
Gel Filtration ◽  
Factor Xa ◽  
Molecular Size ◽  
Molecular Forms ◽  
Factor X ◽  
Monomeric Complex ◽  
At Iii ◽  
Antibody Competition

SummaryA double antibody competition radioimmunoassay was developed that allowed to detect specifically as little as 15 ng antithrombin III (AT III) per ml of the assayed material. In normal plasma examined by this assay, AT III concentration averaged 199 ± 21 μg/ml. Complexes of AT III with thrombin or factor X a crossreacted with free AT III in 87% and 95%, respectively. Molecular forms of AT III produced in plasma treated with coagulation enzymes, or in serum, were assessed by measuring immunoreactive AT III in fractions obtained by gel filtration chromatography on Sephadex G-200. AT III bound by thrombin in fibrinogen free-plasma ranged in molecular size from 160,000 to above 250,000. Similar aggregation occurred when monomeric complex of purified AT III and thrombin, of 90,000 Mr, was added to plasma. Presence of heparin intensified the degree of aggregation. In factor Xa-treated plasma AT III was converted into components with 160,000 Mr, or less. No complexes below 200,000 Mr were present in serum. They decreased in size to 160,000 Mr after affinity chromatography on heparin-Sepharose. These results indicated that blood represents a unique milieu conducive to aggregation of bound AT III. It appears, however, that AT III complexes present in blood may not only aggregate, but also associate with other serum proteins through unstable binding most likely caused by the enzyme component of the complex.

An Assessment Of An Amidolytic Assay For Factor VII In The Laboratory Control Of Oral Anticoagulants

1981 ◽  
Author(s):  
A Bodzenta ◽  
Jean M Thomson ◽  
Z S Latallo
Keyword(s):  
Prothrombin Time ◽  
Oral Anticoagulant ◽  
Oral Anticoagulants ◽  
Factor Vii ◽  
Limiting Factor ◽  
Factor X ◽  
Present Evidence ◽  
Chromogenic Assay ◽  
Better Than

An amidolytic assay for factor VII, modified from the method of Seligsohn et al (1978), has been compared with the results of the prothrombin time using British Comparative Thromboplastin, Thronbotest and a clotting assay for factor VII. In ‘long-term’ oral anticoagulant administration agreement with the conventional methods was good and better than in our previous study when amidolytic assays for factors II and X respectively were studied (Latallo et al 1981). The method appeared to be reasonably specific for factor VII.On the present evidence the chromogenic assay for factor VII offers a limited but apparently dependable guide to dosage but it is elaborate to perform and difficult to standardise. The main limiting factor for its routine application is the need to prepare a purified factor X extract.

Soluble Fibrin Complexes and Fibrinogen Heterogeneity in Diabetes Mellitus

1979 ◽  
Author(s):  
E.B. Tsianos ◽  
N.E. Stathakis
Keyword(s):  
Molecular Weight ◽  
Plasma Proteins ◽  
Antithrombin Iii ◽  
Matched Control ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Lower Molecular Weight ◽  
Soluble Fibrin ◽  
Patients With Diabetes ◽  
Poly Acrylamide

The presence of soluble fibrin complexes (SFC) measured by gel filtration of plasmaon 4% agarose columns, fibrinogen heterogeneity on 3.5% SDS-polyaorylamide gels and the concentration of several plasma proteins were evaluated in 39 patients with diabetes mellituB (DM) and 19 matched control subjects. A small but significant inoreaee of SFC was found in DM (p<0.001). Of the patients 54% had increased concentrations of SFC (>M2SD of the controla). Polyacrylamide gel electrophoresis of the SFC showed no evidence of cross-linking or proteolysis. Plasma olots formed in the presence of EDTA and trasylol were analysed in IDS-poly-acrylamide gels in a normal and two lower molecular weight fibrin bands (band I, II, III). The percentage of band 1 fibrinogen was in diabetics (65.3±4.7%) lower than that of the controls (71.8±4.45%) (p<0.00l). Fibrinogen levels, antithrombin III, ai-antitrypsin, ai-macroglobulin and plasminogen were significantly increased in DM. Patients with increased concentration of SFC had higher concentrations of fibrinogen, ai-antitrypain and band I fibrinogen. We suggest that in DM there is an enhancement of intravaaoular fibrin formation and accelerated fibrinogen degradation to lower molecular weight forms.

scholarly journals Hypoprothrombinemia in the compensated form of hepatosplenic schistosomiasis: further studies

1988 ◽  
Vol 30 (4) ◽  
pp. 274-280 ◽  
Author(s):  
Nora Manoukian ◽  
Durval Rosa Borges
Keyword(s):  
Prothrombin Time ◽  
Plasma Levels ◽  
Antithrombin Iii ◽  
Plasma Concentrations ◽  
Factor Vii ◽  
Low Grade ◽  
Antigen Concentration ◽  
Cirrhotic Group ◽  
Hepatic Synthesis ◽  
Hepatosplenic Schistosomiasis

Coagulation abnormality is frequently observed in schistosomiasis patients but its pathophysiology has not been established. We measured, by immunodiffusion. the prothrombin-antigen concentration in 56 individuals; of these 19 with demonstrated compensated form of hepatosplenic schistosomiasis, 17 with cirrhosis and 20 were control subjects. Transaminases, albumin, transthyretin, prothrombin time, antithrombin III, factor VII, and fibrinogen were also evaluated. All parameters were altered in the cirrhotic group but only albumin, prothrombin and antithrombin III levels were altered in the schistosomiasis group. Ninety percent of the patients with cirrhosis and sixty percent of the patients with schistosomiasis had abnormal plasma levels of albumin, transthyretin, prothrombin-antigen, and/or antithrombin III; an impaired hepatic synthesis was responsible for these results. Conversely forty percent of the schistosomiasis patients with normal plasma concentrations of both albumin and transthyretin had decreased mean plasma levels of both prothrombin and antithrombin III. These results suggest that either proth rombin and antithrombin III are more sensitive markers of impaired hepatic synthesis in schistosomiasis than are levels of albumin and transthyretin combined, or a low grade chronic consumption of clotting proteins also occurs. Considering the latter hypothesis it is possible that the thrombin formed would be inhibited by antithrombin III with the complexed thrombin-antithrombin III being cleared by the liver. Consequently the plasma levels of both prothrombin and antithrombin would be decreased, but the level of fibrinogen would be preserved.

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