Fibrinogen Montreal

1973 ◽  
Vol 29 (03) ◽  
pp. 536-546 ◽  
Author(s):  
M Lacombe ◽  
J Soria ◽  
C Soria ◽  
G d’Angelo ◽  
R Lavallee ◽  
...  
Keyword(s):  
Optical Density ◽  
Polyacrylamide Gel ◽  
Clotting Time ◽  
Functional Defect ◽  
A Chain ◽  
Fibrin Monomers

SummaryA new case of congenital dysfibrinogenemia characterized by a prolonged thrombin clotting time and a low optical density of the polymerization curve has been discovered in Montreal. The functional defect is due to an abnormal aggregation of fibrin monomers.The characteristics of this abnormal fibrinogen are serum gélification (Paracoagulation) at 37°, 22° and 4° C, a normal immuno-electrophoretic and electrofocusing pattern, a slight increase in the mobility in the α (A) chain by electrophoresis of the dissociated chains in polyacrylamide gel. However, no abnormality was found in the α (A) chain of the disulphide knot.

Automation of Two-Stage Factor VIII Assay

1978 ◽  
Vol 39 (02) ◽  
pp. 455-465 ◽  
Author(s):  
Yvonne Stirling ◽  
D J Howarth ◽  
Marguerite Vickers ◽  
W R S North ◽  
T W Meade
Keyword(s):  
Factor Viii ◽  
Optical Density ◽  
Electric Circuit ◽  
Clotting Time ◽  
Two Stage ◽  
Laboratory Error ◽  
Premature Closure ◽  
Factor Viii Concentrates ◽  
Experimental Findings ◽  
Automated Methods

SummaryTwo automated methods for two-stage factor VIII assays have been compared with one another, and evaluated in practice. The Depex method records the clotting time when an electric circuit is completed by the formation of a fibrin thread across a hook-type electrode; the Electra method is based on an optical density technique of clot detection. The two methods gave comparable results for measured levels of factor VIII when haemophilic or “normal” plasmas were assayed. Results from the two methods in practice also suggest that both are valid at low and “normal” factor VIII levels. The Electra method is also probably suitable for assays of concentrates; however, the Depex method appears to give falsely high values in these circumstances, and experimental findings suggest that the reason may be that increased viscosity due to the high fibrinogen levels in factor VIII concentrates causes premature closure of the circuit between the two ends of the Depex electrode. The main advantage of the Depex method is that, provided 3 or 4 machines are available, a given number of assays can be completed more quickly than on Electra. The main advantages of Electra are that it is probably subject to less laboratory error than Depex, and that it is suitable for assaying concentrates as well as haemophilic and “normal” plasmas.

Functional Relationship of Polymerization Sites of Vertebrate Fibrinogens

1979 ◽  
Author(s):  
C Cierniewski ◽  
T Krajewski ◽  
E Janiak
Keyword(s):  
Gel Electrophoresis ◽  
Functional Relationship ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Functional Similarity ◽  
Fractionation Method ◽  
Relationship Of ◽  
Fibrin Monomers ◽  
Fibrinogen Molecule ◽  
Cold Ethanol Fractionation

Various studies on the interaction of immobilized mammalian fibrinogen and fibrin monomers with some fibrinogen derivatives demonstrated the presence of two sets of polymerization sites in the mammalian fibrinogen molecule. We obtained the same results while investigating the fibrinogen molecules of other classes of vertebrates /Pisces. Amphibia. Aves/. Despite significant differences among their subunit structures, all of them contain polymerization sites homologous to mammalian counterparts. Moreover, due to great functional similarity, fibrinogen or fibrin monomers of the analyzed species of Pisces. Amphibia. Aves and Mammalia interacted in a specific way with immobilized pig fibrin monomers or fibrinogen, respectively. Using these pig affinity adsorbents, fibrinogen and fibrin monomers of different vertebrates were isolated directly from plasma and analyzed by SDS polyacrylamide gel electrophoresis. Polypeptide compositions of eluted proteins were identical to those obtained for corresponding fibrinogen preparations isolated by cold-ethanol fractionation method. It appears to indicate that the nature of polymerization sites in vertebrate fibrinogens is alike.

An Abnormal Fibrinogen (Copenhagen) Associated with Severe Thromboembolic Disease, but with Normal Adsorption of Plasminogen

1979 ◽  
Author(s):  
Sandbjerg M. Hansen ◽  
I. Clemmensen
Keyword(s):  
Venous Thrombosis ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Thromboembolic Disease ◽  
Polyacrylamide Gel ◽  
Fibrinopeptide A ◽  
Related Material ◽  
Thrombotic Disease ◽  
Fibrin Formation ◽  
Fibrin Monomers

An autosomally inherited qualitative fibrinogen (F) defect is presented. The abnormal F was detected in the plasma of a 54 year old woman with severe arterial thrombotic disease. A decreased rate of fibrin formation of plasma, or purified F by thrombin or ancrod (Arvin (R ) ) was demonstrated. The plasma F concentration was normal, when estimated by clottability or immunologic technique. No F related material was present in the serum. The purified abnormal F was indistinguishable from normal F by Immunoelectrophoresis or SDS Polyacrylamide gel electrophoresis. The major defects detected were an abnormality of polymerization of fibrin monomers and a decreased rate of release of fibrinopeptide A. To evaluate a possible abnormality of the binding of plasminogen (P) to the abnormal fibrin, we examined the adsorption of partially degraded P (Lys-P), which has a higher affinity for fibrin than Glu-P. The adsorption was normal, but the study might be useful in the evaluation of dysfibrinogenemia associated with venous thrombosis.

Structure Abnormality Of Fibrinogen Metz And Its Relationship To The Clotting Defect

1981 ◽  
Author(s):  
A Henschen ◽  
C Southan ◽  
J Soria ◽  
C Soria ◽  
M Samama
Keyword(s):  
Young Woman ◽  
Sodium Citrate ◽  
Disc Electrophoresis ◽  
Polyacrylamide Gel ◽  
Fibrinopeptide A ◽  
Clotting Time ◽  
Terminal Part ◽  
Fibrin Formation ◽  
No Release ◽  
Α Chain

Fibrinogen Metz was discovered in a young woman and was associated with a mild bleeding disorder and repeated abortions. Her parents are cousins and both have an abnormal fibrin formation but less intense than in the propositus.Fibrinogen Metz is therefore an homozygous case characterized by an unclottability of citrated sample, even in the presence of calcium and large quantities of thrombin. In addition, the clotting time is prolonged when the blood is collected in the absence of sodium citrate and the clot is extremely weak.By immunoenzymological assay, no release of fibrinopeptide A was noted after thrombin addition to purified fibrinogen. Furthermore, by disc electrophoresis in polyacrylamide gel in absence of SDS, an abnormal migration of the A α chain was observed.These results led us to analyze the sequence of the N terminal part of the A α chain by the Edman procedure. Arginine in position 16 was replaced by cysteine in fibrinogen Metz. This mutation localized on the site cleaved by thrombin explains the absence of fibrinopeptide A release, and the abnormal clottability of fibrinogen Metz.

Functional Relationship of Polymerization Sites of Vertebrate Fibrinogens

1979 ◽  
Author(s):  
C.S. Ciernlewski ◽  
T. Krajewski ◽  
E. Janiak
Keyword(s):  
Gel Electrophoresis ◽  
Functional Relationship ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Functional Similarity ◽  
Fractionation Method ◽  
Relationship Of ◽  
Fibrin Monomers ◽  
Fibrinogen Molecule ◽  
Cold Ethanol Fractionation

Various studies on the interaction of immobilized mammalian fibrinogen and fibrin monomers with some fibrinogen derivatives demonstrated the presence of two sets of polymerisation sites In the mammalian fibrinogen molecule. We obtained the same results while investigating the fibrinogen molecules of other classes of vertebrates /Pisces, Amphibia, Aves/. Despite significant differences among their subunit structures, all of them contain polymerization sites homologous to mammalian counterparts. Moreover, due to great functional similarity, fibrinogen or fibrin monomers of the analyzed species of Pisces, Amphibia, Ayes and Mammalia interacted in a specific way with Immobilized pig fibrin monomers or fibrinogen, respectively. Using these pig affinity adsorbents, fibrinogen and fibrin monomers of different vertebrates were isolated directly from plasma and analyzed by SDS Polyacrylamide gel electrophoresis. Polypeptide compositions of eluted proteins were identical to those obtained for corresponding fibrinogen preparations isolated by cold-ethanol fractionation method. It appears to indicate that the nature of polymerization sites in vertebrate fibrinogens is alike.

SEED PROTEIN HOMOLOGIES AND THE EVOLUTION OF POLYPLOIDY IN AVENA

1972 ◽  
Vol 14 (4) ◽  
pp. 875-888 ◽  
Author(s):  
G. Ladizinsky ◽  
B. L. Johnson
Keyword(s):  
Optical Density ◽  
Seed Protein ◽  
Correlation Coefficients ◽  
Disc Electrophoresis ◽  
Polyacrylamide Gel ◽  
Individual Species ◽  
Protein Patterns ◽  
Independent Origin ◽  
Pattern Variability ◽  
Tetraploid Parent

Crude seed protein of the nine oat species was fractionated by disc electrophoresis on polyacrylamide gel. Correlation coefficients calculated from optical density curves obtained from the stained gels showed that individual species possessed characteristic, albeit variable, profiles. Pattern variability among and within species was greater in the case of the diploids than in the case of the polyploids. The very few profile types in A. strigosa 4x and in the hexaploid A. sativa suggested that the variation in these polyploids is due more to independent origin of the types than to differentiation following polyploidization. Virtual identity between individual A. strigosa 2x and 4x profiles suggested a strict autopolyploid origin for some 4x types while complementing pairs of A. strigosa 2x profiles indicated an intervarietal origin for other A. strigosa 4x types. A diagnostic band at 11.0 cm in the profiles of A. magna and A. murphyi suggested that these species rather than A. strigosa 4x had functioned as the tetraploid parent of the hexaploid A. sativa. The profile of A. murphyi complemented by a specific A. strigosa 2x profile simulated a specific A. sativa type. The adaptive success of the genus is assessed in the light of variation and homologies in the seed protein patterns of the various species.

scholarly journals Anticoagulant function of a 24-Kd fragment isolated from human fibrinogen A alpha chains

Blood ◽  
1993 ◽  
Vol 81 (12) ◽  
pp. 3277-3284
Author(s):  
HK Lau
Keyword(s):  
Amino Acid ◽  
Maximum Level ◽  
Polymerization Reaction ◽  
Amino Acid Residues ◽  
Clotting Time ◽  
Human Fibrinogen ◽  
Apparent Homogeneity ◽  
Lag Period ◽  
Anticoagulant Function ◽  
Fibrin Monomers

A fibrinogen fragment obtained by limited-plasmin proteolysis has been isolated and purified to apparent homogeneity by gel filtrations. This fragment, denoted as 24-Kd fragment, has an apparent M(r) approximately 24,000 and contains an N-terminal sequence of met-glu-leu-glu-arg-pro- gly-gly-asn-glu-ile. The fragment contains a large number of acidic amino acid residues, and its amino acid composition is similar to several fibrinogen A alpha chains degradation fragments isolated previously. It corresponds to a peptide of the fibrinogen A alpha chains, the N-terminal of which starts at alpha Met-240. This peptide delays thrombin plasma clotting time. It does not bind calcium ions and does not inhibit thrombin's amidolytic activity. It binds to immobilized fibrin but not fibrinogen. It also inhibits the polymerization of desAA and desAABB fibrin monomers by simultaneously decreasing the maximum rate and the maximum level of the polymerization reaction. However, the initial lag period of this reaction is not affected by the fragment.

Influence of High-dose Aprotinin on Anticoagulation, Heparin Requirement, and Celite- and Kaolin-Activated Clotting Time in Heparin-pretreated Patients Undergoing Open-Heart Surgery

1995 ◽  
Vol 83 (4) ◽  
pp. 679-689. ◽  
Author(s):  
W. Dietrich ◽  
G. Dilthey ◽  
M. Spannagl ◽  
M. Jochum ◽  
S. L. Braun ◽  
...  
Keyword(s):  
Blood Loss ◽  
Thrombin Generation ◽  
Plasma Concentrations ◽  
Postoperative Blood Loss ◽  
High Dose ◽  
Clotting Time ◽  
Activated Clotting Time ◽  
High Dose Aprotinin ◽  
Group A ◽  
Fibrin Monomers

Background Aprotinin causes a prolongation of the celite-activated clotting time (CACT), but not of the kaolin-activated clotting time (KACT). Therefore, concern has been raised regarding the reliability of CACT to monitor anticoagulation in the presence of aprotinin. The current study was designed to test the efficacy of aprotinin to improve anticoagulation, and to investigate whether the prolongation of CACT reflects true anticoagulation or is an in vitro artifact. To elucidate this antithrombotic effect of aprotinin, this study was done in patients prone to reduced intraoperative heparin sensitivity. Methods In a prospective, randomized, double-blind clinical trial, 30 male patients scheduled for elective primary coronary revascularization and treated with heparin for at least 10 days preoperatively, received either high-dose aprotinin (group A) or placebo (group C). The CACT and KACT were determined, but only CACT was used to control anticoagulation with heparin. Parameters of coagulation that are indicators of thrombin generation and activity (F1+2 prothrombin fragments, thrombin-antithrombin III complex, and fibrin monomers), parameters of fibrinolysis (D-dimers), aprotinin, and heparin plasma concentrations were measured. Postoperative blood loss and allogeneic blood transfused were recorded. Results Total heparin administered was 36,200 units (95% confidence interval: 31,400-41,000; group C) compared with 27,700 (25,500-29,800) units (group A; P < 0.05). Hemostatic activation during cardiopulmonary bypass (CPB) was significantly reduced in group A compared with group C. After 60 min of CPB, all parameters were significantly different (P < 0.05) between the groups (group C vs. group A): F1+2 prothrombin fragments, 9.7 (8.9-11.7) ng/ml versus 7.5 (6.2-8.6) ng/ml; thrombin-anti-thrombin III complex (TAT), 53 (42-68) ng/ml versus 29 (23-38) ng/ml; and fibrin monomers, 23 (12-43) ng/ml versus 8 (3-17) ng/ml. Fibrinolysis was also attenuated; D-dimers at the end of operation were 656 (396-1,089) and 2,710 (1,811-4,055) ng/ml for groups A and C, respectively (P < 0.05). The CACT 5 min after the onset of CPB was 552 (485-627) versus 869 (793-955) s for groups C and A, respectively (P < 0.05), whereas the KACT showed no differences between the groups (569 [481-675] vs. 614 [541-697] s for groups C and A, respectively; P = NS). The 24-h blood loss was 1,496 (1,125-1,995) versus 597 (448-794) ml for groups C and A, respectively (P < 0.05). Conclusions Aprotinin treatment in combination with heparin leads to less thrombin generation during CPB. Aprotinin has anticoagulant properties. Celite-activated ACT is reliable for monitoring anticoagulation in the presence of aprotinin, because the prolonged CACT in the aprotinin group reflects improved anticoagulation. Kaolin-activated ACT does not reflect this effect of aprotinin.

Interactions Of Triphenyl-Methane Compounds With Fibrinogen And Fibrin Monomers

1981 ◽  
Author(s):  
J-J Ryckewaert ◽  
R F Doolittle
Keyword(s):  
Binding Sites ◽  
Potent Inhibitor ◽  
Equilibrium Dialysis ◽  
Parent Compound ◽  
Polymerization Process ◽  
Clotting Time ◽  
Binding Ability ◽  
Synthetic Compound ◽  
Fibrin Monomers ◽  
Fibrinogen Molecule

A synthetic compound, aurin tricarboxylic acid (ATA), in the concentration range of 10-100 nanomoles/ml exhibits the property of significantly shortening the thrombin clotting time of fibrinogen. Upon binding to fibrinogen or fibrin monomers, obtained either by the action of thrombin or reptil ase on fibrinogen, ATA possesses the ability to accelerate the polymerization formation of fibrin. Polymerization studies of fibrin monomers at different ionic strengths show that ATA greatly enhances the lateral aggregation of fibrin polymers even at high ionic strength. Calcium ions do not influence the effects of ATA. Spectroscopic methods have been used to study and locate the binding sites in the different parts of the fibrinogen molecule. Compounds with structures that are closely related to that of ATA do not affect the polymerization process, with the exception of the parent compound aurin, which has a slight effect. ATA does not influence the binding ability of fibrinogen for the synthetic peptides: Gly-Pro-Arg and Gly-His-Arg, as was shown by equilibrium dialysis experiments with radioactive- labelled peptides. Nevertheless, in the presence of Gly-Pro-Arg or Gly-Pro-Arg-Pro, which are known to interfere with the polymerization of fibrin monomer, ATA becomes a potent inhibitor of the overall process of fibrin formation. This duality of behavior has permitted us to study the different steps of longitudinal and lateral association of fibrin monomers.

Polymerization Sties Of Desialated Fibrinogen

1981 ◽  
Author(s):  
P M Allison ◽  
N U Bang
Keyword(s):  
Sialic Acid ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Clotting Time ◽  
Mean Values ◽  
Fibrin Monomer ◽  
Sds Page

It has long been known (Laki and Chandrasekhar, Nature, 197:1267,1963) that the enzymatic removal of sialic acid (SA) frcm fibrinogen (F) shortens the thrccabin clotting time. Martinez et al. (J. Lab. Clin. Med., 89:367, 1977) demonstrated that this phenomenon is due to enhanced fibrin monomer polymerization. To more accurately identify the mechanisms involved we examined the binding of normal F, V. cholerae neuraminidase - treated F (NF) and their plasmic digests. Mean SA content of F of 4.5 residues / molecule decreased to unmeasurable levels in NF with a concomitant 10s shortening of the thrombin clotting time. F, NF, and plasmin digests of F (FP) and NF (NFP) were subjected to fibrin-monomer-Sepharose chromatography (FMSC) essentially according to Kudryk and Blcmback (J.Biol. Chan., 249:3322, 1974) and effluent fractions analyzed by SDS polyacrylamide gel electrophoresis (SDS-PAGE) under nonreducing conditions. Mean values for F retained on FMSC and eluted with acid urea buffer was 61%, for NF 86%, for for FP 65%, for NFP 78%. In NFP and FP 3 fragments D (D1, D2 D3) of Mr 103K, 89K, and 83K were discerned by SDS-PAGE. D1 in FP and NFP was almost quantitatively retained on FMSC, whereas little, if any, D3 was retained frcm either digest. FMSC resulted in poor retention of D2 for FP and virtually complete retention of this fragment from NFP. Fragments E were not retained on FMSC of either FP or NFP. The data suggests that desialation of F exposes additional polymerization sites present in D2 but not D3 of F.

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