-
British Journal of Haematology Aug 2002The prothrombin G20210A polymorphism, which correlates with the plasmatic prothombin levels, is the second genetic risk factor for deep venous thrombosis (DVT), although...
The prothrombin G20210A polymorphism, which correlates with the plasmatic prothombin levels, is the second genetic risk factor for deep venous thrombosis (DVT), although its prothrombotic role is mild. Recently, the prothrombin A19911G polymorphism, also associated with slight variations of the prothrombin level, has been suggested to modulate the thrombotic risk of the G20210A polymorphism in a preliminary study including few patients and controls. Our study evaluated the effect of the A19911G polymorphism in the arterial and venous thrombotic risk of the prothrombin 20210G/A genotype, analysing 204 consecutive DVT patients and 204 matched controls. Moreover, we analysed 213 carriers of the 20210G/A genotype (152 with DVT, 26 with arterial thrombosis and 35 healthy subjects) and 10 homozygous 20210 A/A carriers. We developed a simple method to simultaneously determine the genotype of both polymorphisms. In accordance with our case/control study, the A19911G polymorphism did not play a significant role in the development of DVT. Analysis of 120 20210 A alleles demonstrated a complete linkage disequilibrium with the 19911 A allele. These polymorphisms (alone or combined) did not modify the risk of arterial thrombosis. However, the 19911A/G genotype slightly increased the risk of developing DVT in carriers of the 20210G/A genotype (OR 3.34 vs 5.86), supporting that the prothrombin 19911 polymorphism could modulate the risk of the G20210A polymorphism in developing DVT.
Topics: Factor V; Female; Genetic Linkage; Genotype; Humans; Male; Middle Aged; Polymorphism, Genetic; Prothrombin; Risk Factors; Venous Thrombosis
PubMed: 12139755
DOI: 10.1046/j.1365-2141.2002.03624.x -
Thrombosis Research Aug 1999Four-factor PCCs are most frequently used for replacement of vitamin K-dependent clotting factors and inhibitors proteins C and S in patients bleeding after... (Review)
Review
Four-factor PCCs are most frequently used for replacement of vitamin K-dependent clotting factors and inhibitors proteins C and S in patients bleeding after phenprocoumon or warfarin overdose, in vitamin K-deficient patients presenting life-threatening bleeding, and liver disease. Since many of these patients are prone to thromboembolic complications including DIC, all conceivable measures should be taken against the thrombogenic potential of PCC preparations. This thrombogenic potential of PCCs is obviously dependent on several factors including activated clotting factors, lack of inhibitors of blood coagulation, and coagulation factor overload, as well as predisposing factors referred to recipients and drug interactions. The composition of PCC should meet the following criteria: Antithrombin in addition to heparin for the neutralization of FIXa and FXa should be present in the preparations; no overloading with FII and FX; substantially lower FVII than FIX potencies in order to minimize contamination with or generation of FVIIa; and substantial protein C as well as protein S activities. Quality control should include determinations as recommended by the European Pharmacopoeia. Specific assays for quantification of FIXa and FXa are urgently required, and validity of these assays must be proven in surveys. All lots should also be tested for their FVIIa content. Furthermore, the safety of PCCs must be proven by suitable animal models. Whenever possible, patients receiving PCCs should be under low-dose heparin prophylaxis; simultaneous administration of heparin-neutralizing drugs or antifibrinolytic agents must be avoided.
Topics: Blood Coagulation Factors; Coagulation Protein Disorders; Disseminated Intravascular Coagulation; Humans; In Vitro Techniques; Prothrombin; Thrombosis
PubMed: 10499903
DOI: 10.1016/s0049-3848(99)00078-x -
Hepatology (Baltimore, Md.) 1982
Topics: Prothrombin; Vitamin K
PubMed: 7095751
DOI: 10.1002/hep.1840020419 -
Schweizerische Medizinische... Aug 1945
Topics: Blood; Hemostatics; Prothrombin
PubMed: 21008986
DOI: No ID Found -
Thrombosis and Haemostasis Aug 1984Laser nephelometry is a technique which allows the evaluation of the concentration of several serum proteins and clotting factors. By means of this technique it is also... (Comparative Study)
Comparative Study
Laser nephelometry is a technique which allows the evaluation of the concentration of several serum proteins and clotting factors. By means of this technique it is also possible to study the kinetics of the reaction between antigen and antibody. We studied the kinetics of the reaction between prothrombin and an antiprothrombin antiserum using several prothrombins namely: Prothrombin Padua, prothrombin Molise, which are two congenital dysprothrombinemias, cirrhotic, coumarin or normal prothrombins. Different behaviors in the kinetics of the reactions were shown even when the concentration of prothrombins was about the same in all plasma tested. These differences were analyzed by means of a computer (Apple II 48 RAM) programmed to solve four unknown equations (Rodbard's equation). From the data so obtained one can see that when voltages at the beginning and at the end of the reaction are in all cases about the same, a clear difference in the time required to reach half the maximum value of the voltage can still be demonstrated. This parameter, which is expressed in minutes, is longer in coumarin and prothrombin Molise than in controls. On the contrary it is shorter in prothrombin Padua and has about the same value of controls in the cirrhotic patient. Moreover the time at which the maximum rate is obtained is longer in coumarin and prothrombin Molise than in controls and shorter in liver cirrhosis and prothrombin Padua. In conclusion data obtained show that coumarin prothrombin behaves in a different way from cirrhotic prothrombin and also that there is a different behaviour between the two congenital dysprothrombinemias.
Topics: Anticoagulants; Antigen-Antibody Reactions; Humans; In Vitro Techniques; Kinetics; Lasers; Liver Cirrhosis; Nephelometry and Turbidimetry; Prothrombin
PubMed: 6495258
DOI: No ID Found -
Thrombosis Research Sep 2000A common mutation in the prothrombin gene (G20210A) is associated with elevated prothrombin levels and thrombosis. The pathomechanism related to the G20210A mutation is...
A common mutation in the prothrombin gene (G20210A) is associated with elevated prothrombin levels and thrombosis. The pathomechanism related to the G20210A mutation is currently not understood and the interdependence of prothrombin activity and prothrombin concentration in plasma is still poorly defined. Six of 191 blood donors examined in the present study carried the prothrombin allele G20210A. Despite the small number of cases, plasma samples from these individuals had significantly higher prothrombin activities than wild type carriers, whereas their prothrombin concentrations-although elevated-did not differ significantly from wild type. In subjects with the G20210A mutation there was also no significant correlation between prothrombin activity and concentration. Analyzing data from healthy blood donors without the prothrombin G20210A mutation, we found only weak correlations between prothrombin activity and concentration of immunoreactive prothrombin. Samples with a relatively high prothrombin concentration but low activity were observed as well as samples with a relatively high activity for a given concentration (hyperactive prothrombin). F1+2 concentrations as indicators of activated coagulation were only elevated in 13 of 125 investigated samples and could not explain any of these findings. Dysfunctional variants of prothrombin, a well known phenomenon, may be responsible for the former, and we speculate that posttranscriptionally modified prothrombin species may explain the observed functional diversity of this factor including hyperactivity. The genotype-phenotype association of the non-coding G20210A mutation is not clear cut. Therefore, further studies are needed to determine which factors apart from the known G20210A polymorphism regulate prothrombin concentration and/or activity and may trigger the manifestation of thrombosis.
Topics: Adult; Age Factors; Alleles; Antigens; Enzyme-Linked Immunosorbent Assay; Female; Genotype; Heterozygote; Humans; Male; Middle Aged; Peptide Fragments; Point Mutation; Prothrombin; Prothrombin Time; Reference Values; Sex Factors
PubMed: 10974339
DOI: 10.1016/s0049-3848(00)00281-4 -
Clinical and Applied... Apr 2017Recombinant factor VIIa (rFVIIa) is used in the management of bleeding in patients with hemophilia. A generic biosimilar version of NovoSeven is also developed...
Recombinant factor VIIa (rFVIIa) is used in the management of bleeding in patients with hemophilia. A generic biosimilar version of NovoSeven is also developed (AryoSeven). To compare the activation profile of NovoSeven and AryoSeven, 2 commercially available protein complex concentrates (PCCs) were used. Profilnine activated by RecombiPlasTin 2G resulted in conversions of prothrombin to prethrombin and thrombin at 5 to 30 minutes. However, addition of rFVIIa at final concentration range of 0.25 to 0.5 µg/mL to the same mixture resulted in total conversion of prothrombin to thrombin with a doublet at 36 kDa. Recombinant factor VIIa alone did not generate thrombin in native Beriplex, and the addition of rFVIIa to Beriplex failed to generate thrombin. Beriplex activated by RecombiPlasTin 2G resulted in complete conversion of prothrombin to thrombin. Both NovoSeven and AryoSeven exhibited similar activation profiles. These studies indicate that the activation of PCCs by both rFVIIa preparations results in comparable generation of thrombin.
Topics: Blood Coagulation Factors; Factor VIIa; Hemorrhage; Humans; Prothrombin; Recombinant Proteins; Thrombin
PubMed: 27553841
DOI: 10.1177/1076029616663848 -
Thrombosis Research Sep 1987Electroimmunoassay of normal (10-Gla) and Gla-deficient prothrombins containing 0 to 9 Gla (gamma-carboxyglutamyl) residues performed in EDTA against anti- (normal)...
Electroimmunoassay of normal (10-Gla) and Gla-deficient prothrombins containing 0 to 9 Gla (gamma-carboxyglutamyl) residues performed in EDTA against anti- (normal) prothrombin showed that each of the Gla-deficient proteins contained as much antigenic activity as does the normal molecule. In the presence of Ca2+, however, normal prothrombin appeared to show the least while 0- to 2-Gla variants, the most activity. This anomaly arises from the presence of antibodies (Ca-IIAb) which form antigen-antibody (Ag-Ab) complexes when the conformation of the antigen is stabilized in Ca2+. For example, upon fractionation of the antisera and mixing Ca-IIAb (bound to 10-Gla prothrombin, Affi-gel column and eluted by replacing Ca2+ with EDTA) with those reacting with prethrombin1 (non-Gla portion of the molecule), rocket heights in Ca2+ decreased the most with 10- and 9-Gla (32%), followed by 13% with 8-Gla, 2% with 7-Gla and none with 2-Gla prothrombins--indicating that Ca-IIAb do cross react with 7-, 8- and particularly 9-Gla prothrombins. This was also confirmed by the fact that anti 7-Gla but not anti 0-Gla sera contained some antibodies which reacted only in the presence of Ca2+.
Topics: Calcium; Edetic Acid; Glutamates; Glutamic Acid; Immunoassay; Immunochemistry; Prothrombin
PubMed: 2890218
DOI: 10.1016/0049-3848(87)90355-0 -
Thrombosis Research Sep 1987Hybridoma technology was used for the production of murine monoclonal antibodies to bovine normal prothrombin. Hybrid cell cultures were assayed for the production of...
Hybridoma technology was used for the production of murine monoclonal antibodies to bovine normal prothrombin. Hybrid cell cultures were assayed for the production of antibodies, both in the absence and presence of calcium ions, by Enzyme-Linked Immunosorbent Assay (ELISA). Antibody-producing cell lines were cloned two times and grown as ascites tumors. Monoclonal antibodies (McAb), isolated by affinity chromatography (Protein A-Sepharose), were tested for their affinity for normal (10-Gla) and dicoumarol-induced abnormal prothrombins containing 2, 5, 7, 8 and 9 gamma-carboxyglutamyl (Gla) residues. A total of 24 McAb were obtained and the immunoglobulins were of the IgG1 subclass. Nine of the twenty-four McAb did not require Ca2+ for the formation of Ag-Ab complexes, and reacted equally with normal and Gla-deficient prothrombins. These antibodies had affinity for prethrombin1 (P1) but not for the Gla-containing prothrombin fragment1 (F1) portion of the molecule. In contrast, the 15 Ca2+-dependent McAb reacted with F1 but not with P1. They discriminated the abnormal prothrombins based upon their Gla content. For example, though all the Ca2+-dependent McAb formed Ag-Ab complexes with 9-, essentially none formed with 5- or less-Gla prothrombins. [Some reacted equally with 9- and 10-Gla (normal) prothrombin while others had only 25% of normal affinity for 9-Gla isomer]. Only four and twelve of the 15 McAb had some affinity for 7- and 8-Gla variants, respectively. These results show that antibodies which react with the Ca2+-stabilized conformation of prothrombin are not specific for normal prothrombin, as has been reported in the literature.
Topics: Animals; Antibodies, Monoclonal; Antigen-Antibody Reactions; Calcium; Cattle; Glutamates; Glutamic Acid; Hybridomas; Mice; Prothrombin
PubMed: 2890219
DOI: 10.1016/0049-3848(87)90356-2 -
Annals of Internal Medicine Oct 1976
Topics: Blood Coagulation; Disseminated Intravascular Coagulation; Dose-Response Relationship, Drug; Hemophilia A; Humans; Prothrombin
PubMed: 970783
DOI: 10.7326/0003-4819-85-4-531