Fibrinogen Bastia (γ 318 Asp → Tyr) a Novel Abnormal Fibrinogen Characterized by Defective Fibrin Polymerization

1999 ◽  
Vol 82 (12) ◽  
pp. 1639-1643 ◽  
Author(s):  
Karim Chabane Lounes ◽  
Claudine Soria ◽  
Antoine Valognes ◽  
Marie France Turchini ◽  
Jaap Koopman ◽  
...  
Keyword(s):  
Calcium Ions ◽  
Clinical Symptoms ◽  
Degradation Products ◽  
Base Substitution ◽  
Chain Gene ◽  
Fibrinogen Concentration ◽  
Clotting Time ◽  
Terminal Part ◽  
Fibrin Polymerization ◽  
Chain Sequence

SummaryA new congenital dysfibrinogen, Fibrinogen Bastia, was discovered in a 20-year-old woman with no clinical symptoms. The plasma thrombin-clotting time was severely prolonged. The functional plasma fibrinogen concentration was low (0.2 mg/ml), whereas the immunological concentration was normal (2.9 mg/ml). Purified fibrinogen Bastia displayed a markedly prolonged thrombin-clotting time related to a delayed thrombin-induced fibrin polymerization. Both the thrombin-clotting time and the fibrin polymerization were partially corrected by the addition of calcium ions. The anomaly of fibrinogen Bastia was found to be located in the γ-chain since by SDS-PAGE performed according to the method of Laemmli two γ-chains were detected, one normal and one with an apparently lower molecular weight. Furthermore, analysis of plasmin degradation products demonstrated that calcium ions only partially protect fibrinogen Bastia γ-chain against plasmin digestion, suggesting that the anomaly is located in the C-terminal part of the γ-chain. Sequence analysis of PCR-amplified genomic DNA fragments of the propositus demonstrated a single base substitution (G → T) in the exon VIII of the γ chain gene, resulting in the amino acid substitution 318 Asp (GAC) → Tyr (TAC). The PCR clones were recloned and 50% of them contained the mutation, indicating that the patient was heterozygous. These data indicate that residue Asp 318 is important for normal fibrin polymerization and the protective effect of calcium ions against plasmin degradation of the C-terminal part of the γ-chain.

scholarly journals Fibrinogen Marburg: a homozygous case of dysfibrinogenemia, lacking amino acids A alpha 461-610 (Lys 461 AAA-->stop TAA)

Blood ◽  
1992 ◽  
Vol 80 (8) ◽  
pp. 1972-1979 ◽  
Author(s):  
J Koopman ◽  
F Haverkate ◽  
J Grimbergen ◽  
R Egbring ◽  
ST Lord
Keyword(s):  
Disulfide Bonds ◽  
Stop Codon ◽  
Polyclonal Antibodies ◽  
Functional Method ◽  
Terminal Segment ◽  
Plasma Fibrinogen ◽  
Base Substitution ◽  
Chain Gene ◽  
Fibrinogen Concentration ◽  
Alpha Chain

In the A alpha-chain gene coding for an abnormal fibrinogen (fibrinogen Marburg) we identified a single base substitution (A-->T) that changes the codon A alpha 461 AAA (Lys) to TAA (Stop). The propositus was found to be homozygous for the mutation, whereas the father and five siblings were heterozygous, and three other siblings contained only the normal sequence. The stop codon at position 461 results in the deletion of the carboxyl-terminal segment A alpha 461–610. Purified fibrinogen Marburg contained an A alpha-chain with a relative molecular weight of approximately 47,000. The FpA release by thrombin was not affected by this deletion, whereas the fibrin polymerization was strongly decreased. The binding of endothelial cells to immobilized fibrinogen Marburg was almost completely abolished compared with normal fibrinogen. Fibrinogen Marburg contained a substantial amount of albumin linked to the fibrinogen molecule by disulfide bonds, and these fibrinogen-albumin complexes were also present in plasma. The plasma fibrinogen concentration of the propositus was measured by three different methods: a functional method (< 0.25 mg/mL), an immunologic method using polyclonal antibodies (0.6 mg/mL), and an immunologic method based on two monoclonal antibodies specific for the amino- terminus and carboxyl-terminus of the A alpha-chain (< 0.05 mg/mL). Using the two immunologic methods, it appeared that only 10% to 15% of the plasma fibrinogen of the heterozygous siblings was abnormal.

Automated nephelometry of fibrinogen: analytical performance and observations during thrombolytic therapy.

1988 ◽  
Vol 34 (10) ◽  
pp. 2135-2140 ◽  
Author(s):  
J J Hoffmann ◽  
M A Verhappen
Keyword(s):  
Thrombolytic Therapy ◽  
Degradation Products ◽  
Oral Anticoagulants ◽  
Fibrinogen Concentration ◽  
Clotting Time ◽  
Protein Assay ◽  
Laboratory Results ◽  
High Concentrations ◽  
Coagulation Analyzer

Abstract We evaluated the performance of an automated nephelometric determination of fibrinogen, which is an integral part of the prothrombin time assay, in a new coagulation analyzer, the ACL-810 (Instrumentation Laboratory). Results were compared with those by a total clottable protein assay and with the thrombin clotting time assay for fibrinogen. In normal and slightly abnormal plasma, the performance of the ACL method was quite satisfactory (CV 3-10%). However, in abnormal plasma (prolonged prothrombin times because of heparin or oral anticoagulants) the accuracy of the ACL method was poor. Nor could the instrument determine fibrinogen in clearly lipemic plasma. In plasma containing high concentrations of fibrin(ogen) degradation products (FDP), collected during thrombolytic therapy with streptokinase-containing drugs, the ACL method gave spuriously high values for fibrinogen concentration. We determined that this was mainly because of interference by intermediate FDP (fragment Y). Finally, we demonstrated that early FDP (fragment X) increased the ACL results for fibrinogen to the same extent as in the total clottable protein method and that late FDP (fragments D and E) affected the thrombin clotting time method, but not the ACL fibrinogen determination.

A Rapid Method for the Semiquantitative Determination of Fibrinogen and Fibrinogen Degradation Products (FDP) in Defibrination Syndrome

1971 ◽  
Vol 25 (03) ◽  
pp. 555-565 ◽  
Author(s):  
G Sas ◽  
J Jákó ◽  
J Domán ◽  
C László ◽  
J Pádár
Keyword(s):  
Degradation Products ◽  
Substantial Reduction ◽  
Experimental System ◽  
Fibrinogen Concentration ◽  
Thrombin Time ◽  
Clotting Time ◽  
Fibrinogen Degradation Products ◽  
Linear Plot ◽  
Fibrinogen Content

Summary1. It was found that effects of deliberate changes in fibrinogen concentration and in the amount of FDP added to the experimental system (containing pure fibrinogen solution and saline- or serumdiluted plasma) could be approximated with satisfactory accuracy by a linear plot of the logarithm of clotting times versus the inverse of fibrinogen concentration.By increasing FDP activity the slope of the obtained lines becomes proportionately steeper. The constants, which interrelate clotting time, fibrinogen concentration and FDP activity, are to be derived experimentally. The obtained formula is expressed also nomographically.2. Apart from the presence of some very rare anticoagulants, an elongation of thrombin time observed under strictly specified conditions points to a substantial reduction of fibrinogen concentration and/or an interplay of fibrinogen degradation products.If a definite amount of fibrinogen (Fibrinogen sec. Warner Chilcott) is admixed to a pathological plasma, thrombin time in the latter will decrease in a specifiable manner. By entering the original and corrected thrombin time values in the reported nomogram the fibrinogen content and FDP activity of the pathological plasma can be calculated.3. The described procedure for fibrinogen and FDP assay is suitable first of all in acute defibrination syndrome and at the thrombolytic therapy. Its agreement with the results obtained by immunodiffusion was satisfactory as regards the fibrinogen in plasmas of different fibrinogen concentration.

Normal Binding of Calcium to Five Fibrinogen Variants with Mutations in the Carboxy Terminal Part of the γ-Chain

1996 ◽  
Vol 76 (03) ◽  
pp. 377-383 ◽  
Author(s):  
Miha Furlan ◽  
Bettina Stucki ◽  
Colette Steinmann ◽  
Myriam Jungo ◽  
Bernhard Lämmle
Keyword(s):  
Calcium Ions ◽  
Calcium Binding ◽  
Single Amino Acid ◽  
Thrombin Time ◽  
Terminal Part ◽  
Calcium Binding Site ◽  
Fibrin Polymerization ◽  
High Affinity ◽  
Carboxy Terminal ◽  
Fibrin Monomers

SummaryCalcium ions are known to accelerate polymerization of fibrin monomers. Each of the two carboxy terminal domains of normal fibrinogen contains one high-affinity calcium binding site that seems to be situated close to the polymerization site in the γ-chain. Most hitherto described functionally defective fibrinogen variants showed impaired clot formation. Since the tightly bound calcium ions may influence the conformation of the polymerization site, the question arises whether the abnormal clotting of a dysfibrinogen might be due to defective calcium binding. We investigated binding of calcium to fibrinogen and the effect of calcium on the clotting properties of five heterozygous fibrinogen variants showing normal thrombin-induced fibrinopeptide release but abnormal polymerization of fibrin monomers. Each of these dysfibrinogens has one single amino acid substitution in the carboxy-terminal part of the γ-chain: fibrinogen Claro (γ 275 Arg ⟶ His), Milano V (γ 275 Arg Cys), Milano I (γ 330 Asp ⟶ Val), Bern I (γ 337 Asn Lys), and Milano VII (γ 358 Ser Cys). The shortest thrombin clotting time and the earliest onset of turbidity increase were observed in fibrinogen γ 358 Ser ⟶ Cys; both parameters were little affected by calcium concentration. In the variant γ 337 Asn ⟶ Lys, the thrombin time was abnormally prolonged at 0.01 mM Ca2+, but it was normalized at 1 mM calcium. In contrast, the abnormal fibrin polymerization of fibrinogen γ 330 Asp ⟶ Val was barely improved at increasing calcium concentrations. Both variants with the substitution of γ 275 Arg, the residue indispensable for normal D:D interactions, showed the slowest rate of fibrin polymerization and the lowest turbidity of fibrin clots at any Ca2+ concentration used. High affinity calcium binding was found to be normal in all five fibrinogen variants studied, suggesting that their abnormal clotting was not due to defective binding of calcium. The γ-chain in the fragment D1 derived from the variant γ 337 Asn ⟶ Lys was further degraded by plasmin in the presence and in the absence of calcium, whereas fragments D1 from the other four γ-chain variants as well as from normal fibrinogen were protected against plasmic degradation in the presence of 1 mM Ca2+.

scholarly journals Fibrinogen Marburg: a homozygous case of dysfibrinogenemia, lacking amino acids A alpha 461-610 (Lys 461 AAA-->stop TAA)

Blood ◽  
1992 ◽  
Vol 80 (8) ◽  
pp. 1972-1979 ◽  
Author(s):  
J Koopman ◽  
F Haverkate ◽  
J Grimbergen ◽  
R Egbring ◽  
ST Lord
Keyword(s):  
Disulfide Bonds ◽  
Stop Codon ◽  
Polyclonal Antibodies ◽  
Functional Method ◽  
Terminal Segment ◽  
Plasma Fibrinogen ◽  
Base Substitution ◽  
Chain Gene ◽  
Fibrinogen Concentration ◽  
Alpha Chain

Abstract In the A alpha-chain gene coding for an abnormal fibrinogen (fibrinogen Marburg) we identified a single base substitution (A-->T) that changes the codon A alpha 461 AAA (Lys) to TAA (Stop). The propositus was found to be homozygous for the mutation, whereas the father and five siblings were heterozygous, and three other siblings contained only the normal sequence. The stop codon at position 461 results in the deletion of the carboxyl-terminal segment A alpha 461–610. Purified fibrinogen Marburg contained an A alpha-chain with a relative molecular weight of approximately 47,000. The FpA release by thrombin was not affected by this deletion, whereas the fibrin polymerization was strongly decreased. The binding of endothelial cells to immobilized fibrinogen Marburg was almost completely abolished compared with normal fibrinogen. Fibrinogen Marburg contained a substantial amount of albumin linked to the fibrinogen molecule by disulfide bonds, and these fibrinogen-albumin complexes were also present in plasma. The plasma fibrinogen concentration of the propositus was measured by three different methods: a functional method (< 0.25 mg/mL), an immunologic method using polyclonal antibodies (0.6 mg/mL), and an immunologic method based on two monoclonal antibodies specific for the amino- terminus and carboxyl-terminus of the A alpha-chain (< 0.05 mg/mL). Using the two immunologic methods, it appeared that only 10% to 15% of the plasma fibrinogen of the heterozygous siblings was abnormal.

Fibrin Polymerization Defect in HIV-infected Patients-Evidence for a Critical Role of Albumin in the Prolongation of Thrombin and Reptilase Clotting Times

1995 ◽  
Vol 73 (03) ◽  
pp. 349-355 ◽  
Author(s):  
Pierre Toulon ◽  
Elyane Frere ◽  
Claude Bachmeyer ◽  
Nathalie Candia ◽  
Philippe Blanche ◽  
...  
Keyword(s):  
Critical Role ◽  
Human Albumin ◽  
Albumin Level ◽  
Final Concentration ◽  
Albumin Concentration ◽  
Clotting Time ◽  
Fibrin Polymerization ◽  
Small Series ◽  
Group 1

SummaryThrombin clotting time (TCT) and reptilase clotting time (RCT) were found significantly prolonged in a series of 72 HIV-infected patients drawn for routine coagulation testing. Both TCT and RCT were highly significantly correlated with albumin (r = -0.64, and r = -0.73 respectively, p<0.0001). TCT and RCT were significantly higher (p<0.0001) in a series of 30 other HIV-infected patients selected on their albumin level below 30.0 g/l (group l) than in 30 HIV-infected patients with albumin level above 40.0 g/l or in 30 HIV-negative controls; the two latter groups were not different. In vitro supplementation of plasma from group 1 patients with purified human albumin up to 45.0 g/l (final concentration) lead to a dramatic shortening effect on both TCT and RCT, which reached normal values. The TCT and RCT of the purified fibrinogen solutions (2.0 g/l final concentration) were not different in the three groups, and normal polymerization curves were obtained in all cases. This further ruled out the presence of any dysfibrinogenemia in the plasma from group 1 patients. Using purified proteins, highly significant correlations were demonstrated between the albumin concentration and the prolongations of both TCT and RCT, which were of the same magnitude order than those found in the patients plasma. These results suggest that hypo-albuminemia is responsible for the acquired fibrin polymerization defect reported in HIV-infected patients. The pathophysiological implication of the low albumin levels was suggested by the finding of decreased albumin levels (associated with prolonged TCT and RCT) in a small series of the eight HIV-infected patients who developed thrombotic complications.

scholarly journals An Evaluation of Thrombotic Tendency by Whole-Body Enhanced CT Scan for Critical COVID-19 Pneumonia: A Case Series Study

2021 ◽  
Author(s):  
Fumihiro Ogawa ◽  
Yasufumi Oi ◽  
Kento Nakajima ◽  
Reo Matsumura ◽  
Tomoki Nakagawa ◽  
...  
Keyword(s):  
Respiratory Failure ◽  
Ct Scan ◽  
Clinical Symptoms ◽  
Degradation Products ◽  
Thrombus Formation ◽  
Case Series ◽  
Whole Body ◽  
Pathological Feature ◽  
Case Series Study ◽  
Enhanced Ct

Abstract Background: Coronavirus disease (COVID-19) pneumonitis associated with severe respiratory failure has a high mortality rate. Based on recent reports, the most severely ill patients present with coagulopathy, and disseminated intravascular coagulation (DIC)-like massive intravascular clot formation is frequently observed. Coagulopathy has emerged as a significant contributor to thrombotic complications. Although recommendations have been made for anticoagulant use for COVID-19, no guidelines have been specified.Case presentation: We describe four cases of critical COVID-19 with thrombosis detected by enhanced CT scan. The CT findings of all cases demonstrated typical findings of COVID-19 and pulmonary embolism or deep venous thrombus without critical exacerbation. Two patients died of respiratory failure due to COVID-19.Discussion: Previous reports have suggested coagulopathy with thrombotic signs as the main pathological feature of COVID-19, but no previous reports have focused on coagulopathy evaluated by whole-body enhanced CT scan. Changes in hemostatic biomarkers, represented by an increase in D-dimer and fibrin/fibrinogen degradation products, indicated that the essence of coagulopathy was massive fibrin formation. Although there were no clinical symptoms related to their prognosis, critical COVID-19-induced systemic thrombus formation was observed. Conclusions: Therapeutic dose anticoagulants should be considered for critical COVID-19 because of induced coagulopathy, and aggressive follow-up by whole body enhanced CT scan for systemic venous thromboembolism (VTE) is necessary.

THE PROLONGATION OF THE THROMBOTEST CLOTTING TIME IN NEWBORNS

1987 ◽  
Author(s):  
K Hamulyák ◽  
P P Devilée ◽  
W Nieuwenhulzen ◽  
H C Hemker
Keyword(s):  
Umbilical Cord ◽  
Vitamin K ◽  
Degradation Products ◽  
Coagulation Factors ◽  
Human Umbilical Cord ◽  
Clotting Time ◽  
Fibrinogen Degradation Products ◽  
Cord Plasma ◽  
Dilution Curves

We investigated 40 human umbilical cord plasma samples of healthy full term infants. The samples were drawn under conditions which minimize in vitro activation of the haemostatic mechanism. In 67% clotting inhibiting material was present as judged from thrombotest dilution curves. The prothrombin (factor II) clotting activity (one stage method) ranged from 28 to 74% (mean value 50.6% ± 12.1). The correlation coefficient between the thrombotest clotting times and theprothrombin levels was -0.46. Using sensitive immuno assays for fibrin degradation products (XDP) and fibrinogen degradation products (FDP) based on monoclonal antibodies, we found that no degradation products could be demonstrated in the non-inhibited group (in the thrombotest dilution curves) whereas small amounts of these products were present in the inhibited group. These small amounts were undetectable using conventional assays. The most striking finding was the presence of fibrin and fibrinogen degradation products. The prolongation of the thrombotest clotting time could be imitated by adding small amounts of purified degradation product fragment X to umbilical cord plasma with a normal thrombotest clotting time. The thrombotest clotting time is often used in clinical practice to obtain an impression of the levels of the vitamin K-dependent coagulation factors. We conclude that the thrombotest clotting time is of limited value in the assessment of the vitamin K-dependent coagulation factors in umbilical cord plasma because in 67% of the cases a further ("false") prolongation of the thrombotest clotting time is caused by small amounts of fibrin(ogen) degradation products. As utmost care was taken to avoid proteolytic breakdown in vitro, our findings most likely reflect an enhanced fibrino(geno)lytic activity in umbilical cord plasma in vivo.

Effects of intravenous administration of a fat emulsion on blood coagulation in dogs

1959 ◽  
Vol 196 (5) ◽  
pp. 1015-1019 ◽  
Author(s):  
Harrison H. Shoulders ◽  
Robert C. Hartmann ◽  
H. C. Meng
Keyword(s):  
Blood Coagulation ◽  
Intravenous Administration ◽  
Intravenous Infusion ◽  
Bleeding Time ◽  
Plasma Fibrinogen ◽  
Fibrinogen Concentration ◽  
Clotting Time ◽  
Clot Retraction ◽  
Fat Emulsion ◽  
Abnormal Bleeding

A fat emulsion prepared for intravenous administration has been studied with regard to its effect upon blood coagulation in dogs. The most characteristic effects found during intravenous infusion of this material at a rate of 1 ml/min. were thrombocytopenia and marked shortening of clotting time. These effects were invariably observed in animals which had not previously received the emulsion. When subsequent infusions were given within 3 hours, no significant changes were observed. When the interval was extended to 1–13 days, variable responses occurred, at times characterized by pronounced hypocoagulability. If the repeat infusion was given 2 weeks or more after the initial one, the effects were similar to those observed during the first infusion. The prothrombin and thrombin clotting times and plasma fibrinogen concentration were not greatly altered during the infusion. Abnormal bleeding time, ‘prothrombin utilization’ and clot retraction accompanied the thrombocytopenia.

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