DETECTION OF SOLUBLE FIBRIN MONOMER COMPLEXES - COMPARISON OF A HAEMAGGLUTINATION ASSAY WITH THE ETHANOL GELATION TEST

1987 ◽  
Author(s):  
G Oehler ◽  
H Klaus ◽  
E Spanuth ◽  
K E Stötzer
Keyword(s):  
Degradation Products ◽  
Positive Test ◽  
Test Results ◽  
Soluble Fibrin ◽  
Fibrin Degradation Products ◽  
Fibrin Monomer ◽  
At Iii ◽  
Positive Results ◽  
Fibrin Monomers ◽  
Gelation Test

Hypercoagulability and disseminated intravascularcoagulation (DIC) are characterized by the presenceof circulating fibrin monomer complexes in plasma.In342 patients with possible DIC fibrin monomers, fibrinogen, reptilase time, antithrombin III and othercoagulation parameters were determined at frequent intervals.Testing of soluble fibrin monomer complexeswas performed using a sensitive and reliable haemagglut- ination assay, with red cells sensitized by fibrin monomers (FM-Test) and the ethanol gelation test(EGT). Method comparison regarding the influence offibrinogen levels and fibrin degradation products shows that high fibrinogen levels lead to false positive results with EGT. The same effect is observed forfibrin degradation products and EGT whereas no influence of fibrinogen level and fibrin degradation products on the FM-Test occurs.It could be shown that with normal fibrinogen concentrations (200-400 mg/dl) the positive test results by FMT and EGT are comparable, whereas with fibrinogen concentrations below 200 mg/dl the number of positive results obtained with the EGT amounted to half the number given by FMT. In the case of fibrinogen concentrations above 400 mg/dl, positive results obtained with EGT were 3.3 times higher than FMT. Nearlyidentical results were obtained by comparing the influence of degradation products. In case of high degradation product concentrations, EGT gives 4.5 timesmore positive results than FMT.Further we compared the number of positive test results obtained by the FMT with the level of AT III because it is wellknown that the AT IIIHevel decreases caused by proteolytic activity generated in DIC.In this study it could be shown that fibrin monomer increases in parallel with the decrease of AT III. Thiseffect does not occur with fibrin degradation products.

The Resistance of Soluble Derivatives of Fibrinogen and the Sensitivity of Its Insoluble Forms to Fibrinolytic Degradation in Blood

1974 ◽  
Vol 32 (02/03) ◽  
pp. 582-591 ◽  
Author(s):  
Victor Gurewich ◽  
Andrzej Nowak ◽  
Izabella Lipinska ◽  
Boguslaw Lipinski
Keyword(s):  
Fibrinolytic Activity ◽  
Degradation Products ◽  
Venous Occlusion ◽  
Protamine Sulfate ◽  
Electrophoretic Method ◽  
Soluble Fibrin ◽  
Fibrin Degradation Products ◽  
Fibrin Monomer ◽  
Derivatives Of

SummaryThe effect of naturally induced fibrinolytic activity on fibrinogen and certain soluble and insoluble derivatives was studied. Experiments were performed on blood removed after venous occlusion of the arm and immediately after death. A previously described electrophoretic method was used by which the heterogeneity of fibrinogen can be demonstrated directly in intact plasma. It was shown that fibrinogen, soluble fibrin monomer (FM) complexes and fibrin degradation products are resistant to degradation by naturally-induced fibrinolytic activity. By contrast, rapid lysis of fibrin, protamine sulfate (PS) precipitated fibrinogen, and PS and ethanol induced gels of FM occurred. The observations are believed relevant to our understanding of the pathway of fibrinogen and FM catabolism and the interpretation of the origin of serum FDP.

Measurement of soluble fibrin monomer-fibrinogen complex in plasmas derived from patients with various underlying clinical situations

2003 ◽  
Vol 89 (05) ◽  
pp. 832-836 ◽  
Author(s):  
Yumiko Kazahaya ◽  
Yuichi Shintani ◽  
Kensuke Yamazumi ◽  
Yutaka Eguchi ◽  
Shin Koga ◽  
...  
Keyword(s):  
Molecular Markers ◽  
Disseminated Intravascular Coagulation ◽  
Degradation Products ◽  
Research Society ◽  
Distinct Entity ◽  
Soluble Fibrin ◽  
Intravascular Coagulation ◽  
Fibrin Degradation Products ◽  
Fibrin Monomer ◽  
D Dimer

SummaryWe previously reported a monoclonal antibody named IF-43 that specifically recognizes thrombin-modified fibrinogen (desAA- and desAABB- fibrin monomer) bound with fibrinogen or other D1 domain-containing plasmic fragments such as fragments X, Y, and D1, but not intact fibrinogen or cross-linked fibrin degradation products (XDP). Here, we tentatively named such complexes, soluble fibrin monomer (FM) -fibrinogen complex.By utilizing IF-43, we have developed a kit to measure soluble FM-fibrinogen complex and compared the profiles with those of two established molecular markers for thrombo-embolic disorders: i.e. the thrombin-antithrombin complex (TAT) and the D-dimer in plasma of patients who underwent surgery without any thrombo-embolic complications. The result indicated that soluble FM-fibrinogen complex is a distinct entity from the two established molecular markers. We have also attempted to observe their profiles in patients with the disseminated intravascular coagulation syndrome (DIC). Although the profiles of soluble FM-fibrinogen complex in individual patients appeared to vary from one patient to the other, the plasma level of soluble FM-fibrinogen complex was found to be increased at the initial phase of disseminated intravascular coagulation syndrome. Thus, the soluble FM-fibrinogen complex may serve as an independent molecular marker for the detection of thrombin generation and the diagnosis of thrombosis. The soluble FM-fibrinogen complex may also serve as a risk factor for thrombosis, because it may precipitate as insoluble complexes beyond its threshold in plasma, or when it is modified by thrombin.Part of this paper was originally presented at the 17th International Fibrinogen Workshop of the International Fibrinogen Research Society (IFRS) held in Munich, Germany, September, 2002.

Comparison of Ethanol and Protamine Tests in Demonstration of Soluble Fibrin and Early Products of Fibrin Degradation

1972 ◽  
Vol 28 (03) ◽  
pp. 342-350 ◽  
Author(s):  
Y. P Konttinen ◽  
L Kemppainen ◽  
O Turunen
Keyword(s):  
Spectrophotometric Method ◽  
Degradation Products ◽  
The Other ◽  
Clot Lysis ◽  
Fibrinogen Degradation Products ◽  
Soluble Fibrin ◽  
Intravascular Coagulation ◽  
Fibrin Monomer ◽  
Other Hand ◽  
Gelation Test

SummaryPerformance and applicability of ethanol-induced gelation and protamine-induced paracoagulation for the demonstration of soluble fibrin monomer complexes and fragment Xo complexes was studied by using 1. fibrin monomer plasma prepared by adding small amounts of thrombin to plasma, 2. clot lysis products, and 3. thrombin -treated mixture of fibrinogen degradation products and plasma. To increase the specificity of the protamine tests only visible fibrin strand formation was recorded as positive. In addition to qualitative tests the amount of paracoagulable material was measured by a spectrophotometric method.The ethanol gelation test proved very simple, reproducible and considerably more sensitive than the protamine tests in demonstrating soluble fibrin monomer complexes, irrespective of whether fibrinogen degradation products were present or not. On the other hand, the protamine tests were clearly superior for demonstration of clot lysis products (fragment Xo complexes). Therefore it seems advisable to perform both types of tests when screening for intravascular coagulation.

scholarly journals Detection of soluble intermediates of the fibrinogen-fibrin conversion using erythrocytes coated with fibrin monomers

Blood ◽  
1976 ◽  
Vol 47 (6) ◽  
pp. 991-1002 ◽  
Author(s):  
R Largo ◽  
V Heller ◽  
PW Straub
Keyword(s):  
Human Plasma ◽  
Incubation Time ◽  
Degradation Products ◽  
Plasma Fibrinogen ◽  
Fibrinogen Concentration ◽  
Agglutination Reaction ◽  
Soluble Fibrin ◽  
Fibrin Degradation Products ◽  
Erythrocyte Concentration ◽  
Fibrin Monomers

Abstract The presence of minimal amounts of fibrinogen-fibrin intermediates in human plasma was visualized by an agglutination reaction of glutaraldehyde-treated human erythrocytes coated with purified fibrin monomers. A degree of monomer coating was established which produced erythrocytes not agglutinated by normal plasma but by plasma containing minimal amounts of soluble complexes of fibrinogen with fibrin monomers. Under standardized conditions of coating, erythrocyte concentration, temperature, pH, and incubation time, the agglutination time varied with the ratio of soluble fibrin to fibrinogen in plasma. The test was sensitive down to a soluble fibrin concentration of 0.675% of the plasma fibrinogen concentration. Early fibrinogen and fibrin degradation products (FDP) in the plasma led to a prolongation of the agglutination time at a concentration of more than 16 mg/100 ml. Late FDP in a concentration of 100 mg/100 ml did not convert a positive test to negative. The test was not affected by heparin and protamine at concentrations of up to 12.5 and 50 NIH units/ml, respectively.

scholarly journals Detection of soluble intermediates of the fibrinogen-fibrin conversion using erythrocytes coated with fibrin monomers

Blood ◽  
1976 ◽  
Vol 47 (6) ◽  
pp. 991-1002
Author(s):  
R Largo ◽  
V Heller ◽  
PW Straub
Keyword(s):  
Human Plasma ◽  
Incubation Time ◽  
Degradation Products ◽  
Plasma Fibrinogen ◽  
Fibrinogen Concentration ◽  
Agglutination Reaction ◽  
Soluble Fibrin ◽  
Fibrin Degradation Products ◽  
Erythrocyte Concentration ◽  
Fibrin Monomers

The presence of minimal amounts of fibrinogen-fibrin intermediates in human plasma was visualized by an agglutination reaction of glutaraldehyde-treated human erythrocytes coated with purified fibrin monomers. A degree of monomer coating was established which produced erythrocytes not agglutinated by normal plasma but by plasma containing minimal amounts of soluble complexes of fibrinogen with fibrin monomers. Under standardized conditions of coating, erythrocyte concentration, temperature, pH, and incubation time, the agglutination time varied with the ratio of soluble fibrin to fibrinogen in plasma. The test was sensitive down to a soluble fibrin concentration of 0.675% of the plasma fibrinogen concentration. Early fibrinogen and fibrin degradation products (FDP) in the plasma led to a prolongation of the agglutination time at a concentration of more than 16 mg/100 ml. Late FDP in a concentration of 100 mg/100 ml did not convert a positive test to negative. The test was not affected by heparin and protamine at concentrations of up to 12.5 and 50 NIH units/ml, respectively.

Recent Advances In The Study Op The Mechanism Op Fierin Polymerization

1981 ◽  
Author(s):  
Stephanie A Olexa
Keyword(s):  
Degradation Products ◽  
Three Dimensional ◽  
Gamma Chain ◽  
Covalent Crosslinking ◽  
Fibrin Degradation Products ◽  
Fibrin Network ◽  
D Region ◽  
Fibrin Monomers ◽  
Fibrinogen Molecule ◽  
Complementary Binding

Fibrinogen is converted to fibrin monomer by thrombin, which cleaves the fibrinopeptides A and E. The resulting fibrin monomers spontaneously polymerize to form a fibrin network, which is later stabilized with covalent crosslinking bonds. Although the action of thrombin and Factor XHIa have been fairly well defined, the mechanism of fibrin polymerization is not yet understood. Electron microscopy and light scattering studies have provided information on the arrangement of molecules within the fiber. Monomers appear to align with a half-staggered overlap, resulting in a fiber with a width equal to twice that of fibrinogen. Detailed studies on the location and interaction of polymerization sites have been done primarily by using fibrinogen and fibrin degradation products. The studies indicate that polymerization is due to the interaction of complementary binding sites which are located on the Fragment D and the NH2- terminal domains in the fibrinogen molecule. One of these sites, located in the sequence 373410 of the gamma chain, is available on fibrinogen in the Fragment D domain. This site is complementary to the site revealed by the loss of fibrinopeptide A, which is probably located in the sequence contiguous to PPA. A second polymerization site in the NH2- terminal domain of fibrin may be located in the sequence following fibrinopeptide B. The recriprocal site may also be located on the Fragment D region. The polymerization of fibrin appears to be due to a complex interaction between molecules with both the primary sequence and the three-dimensional conformation having vital roles.

scholarly journals D-Dimer and Fibrin Degradation Products Impair Platelet Signaling: Plasma D-Dimer Is a Predictor and Mediator of Platelet Dysfunction During Trauma

2020 ◽  
Vol 5 (6) ◽  
pp. 1253-1264
Author(s):  
Christopher C Verni ◽  
Antonio Davila ◽  
Carrie A Sims ◽  
Scott L Diamond
Keyword(s):  
Degradation Products ◽  
Calcium Mobilization ◽  
Factor Xiiia ◽  
Platelet Glycoprotein ◽  
Trauma Patients ◽  
Platelet Dysfunction ◽  
Soluble Fibrin ◽  
Glycoprotein Vi ◽  
Fibrin Degradation Products ◽  
D Dimer

Abstract Background Platelet dysfunction often accompanies trauma-induced coagulopathy. Because soluble fibrin impairs platelet glycoprotein VI (GPVI) signaling and platelets of trauma patients can display impaired calcium mobilization, we explored the role of fibrinolysis on platelet dysfunction during trauma. Methods Convulxin-induced GPVI calcium mobilization was investigated in healthy platelet-rich plasma (PRP) pretreated with thrombin and tissue plasminogen activator (tPA). Blood samples from healthy participants (n = 7) and trauma patients (n = 22) were tested for platelet calcium mobilization, plasma D-dimer, platelet D-dimer binding (via flow cytometry), and platelet lumi-aggregometry. Results For healthy platelets, maximal platelet dysfunction was observed when cross-linked soluble fibrin (no tPA) or cross-linked fibrin degradation products (FDPs) were generated in suspension before convulxin stimulation. Lack of fibrin polymerization (inhibited by Gly-Pro-Arg-Pro [GPRP]) or lack of factor XIIIa cross-linking (T101-inhibited) restored GPVI signaling, whereas non–cross-linked FDPs only partially blocked signaling induced by convulxin. In addition, D-dimer added to healthy PRP impaired platelet aggregation and dense granule release induced by various agonists. Plasma D-dimer level was strongly correlated (R = 0.8236) with platelet dysfunction as measured by platelet calcium mobilization induced with various agonists. By 48 to 120 h after trauma, plasma D-dimer levels declined, and platelet function increased significantly but not to healthy levels. Trauma platelets displayed elevated D-dimer binding that was only partially reduced by αIIbβ3-inhibitor GR144053. After 60-minute incubation, washed healthy platelets resuspended in plasma from trauma patients captured approximately 10 000 D-dimer equivalents per platelet. Conclusions During trauma, D-dimer and FDPs inhibit platelets, potentially via GPVI and integrin αIIbβ3 engagement, contributing to a fibrinolysis-dependent platelet loss-of-function phenotype.

scholarly journals Production and characterization of a monoclonal antibody reactive with a specific neoantigenic determinant (comprising B beta 54-118) in degradation products of fibrin and of fibrinogen

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 437-441 ◽  
Author(s):  
PW Koppert ◽  
J Koopman ◽  
F Haverkate ◽  
W Nieuwenhuizen
Keyword(s):  
Degradation Products ◽  
Blood Clot ◽  
Immunoaffinity Chromatography ◽  
Tissue Type ◽  
Spleen Cells ◽  
Fibrin Degradation Products ◽  
Type Plasminogen Activator ◽  
Complete Lysis ◽  
Fibrin Monomers

Abstract Balb/c mice were immunized with a mixture of fibrin degradation products (XDPs) prepared by complete lysis of a human blood clot by tissue-type plasminogen activator and purified by immunoaffinity chromatography. Spleen cells of the mice were fused with P3 X 63 Ag 8653 myeloma cells. A clone (FDP 14) was selected that produces monoclonal antibodies (MoAbs) of the IgG1 kappa type that react with a neoantigenic determinant exposed in these XDPs, but not in intact fibrinogen or in fibrin monomers. Furthermore, the MoAb is reactive with some pure, individual degradation products of fibrinogen (fragments X, Y, E, and the N-terminal disulphide knot) and with the fibrinogen B beta-chain but not with A alpha- and gamma-chains or with fragments D, FCB-2 and FCB-3. Comparison of the known primary structures of these fibrinogen fragments indicates that the stretch B beta 54–118 comprises at least an important part of the epitope recognized by FDP-14. Apparently, this stretch contributes importantly to a neoantigenic determinant that is not functional in intact fibrinogen and fibrin monomer and that can be made functional by reduction of fibrinogen, or by digestion with plasmin or CNBr.

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