Hereditary Hypodysfibrinogenemia with Defective Release of Fibrinogenopeptide A (Fibrinogen Freiburg)

1979 ◽  
Author(s):  
D. Böttcher ◽  
K. Hasler ◽  
E. Köttgen ◽  
J. Maurath
Keyword(s):  
Bleeding Episode ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Thromboembolic Disease ◽  
Thrombin Time ◽  
Excessive Bleeding ◽  
Major Bleeding Episode ◽  
History Of ◽  
Fibrin Monomers ◽  
Major Defect

A new autoaomally inherited hypodysfibrinogenemia was recognized in four members in three different generations of a family. Only one patient had a major bleeding episode after trauma, the other affected members had no history of excessive bleeding or thromboembolic disease.The thrombin time and Reptilase time of plasma were greatly prolonged and partially corrected by the addition of calcium. Patient plasma prolonged the thrombin time of normal plasma. Fibrinogen levels ranged between 10 to 20 mg/100ml when measured as thrombin-clottable protein, whereas immunologically the fibrinogen levels were only slightly reduced.Functionally the major defect was impaired release of fibrinogenopeptide A upon incubation of the purified abnormal fibrinogen (94% clottable protein) with thrombin and Reptilase. The abnormal fibrinogen showed a delayed polymerisation of its purified fibrin monomers. The described abnormal fibrinogen was indistinguishable from normal fibrinogen by polyacrylamide gel electrophoresis with and without sodium dodecyl sulfate.

Fibrinogen New Orleans: An Inherited Variant with Abnormal Peptide Release

1979 ◽  
Author(s):  
S.I. Chavin ◽  
W.A. Andes ◽  
W.G. Beltran ◽  
W.J. Stuckey
Keyword(s):  
Laboratory Finding ◽  
Polyacrylamide Gel Electrophoresis ◽  
Complete Fusion ◽  
Thrombin Time ◽  
Clotting Time ◽  
Excessive Bleeding ◽  
Normal Limits ◽  
Peptide Release ◽  
Mother And Daughter ◽  
History Of

An inherited fibrinogen abnormality is described in a 30-year old woman with a history of several episodes of excessive bleeding. The initial laboratory finding was a prolonged thrombin time, and a comparable prolongation was present in the plasmas of her mother and daughter, but not of her father. The abnormal fibrinogen gave a reaction of complete fusion with normal fibrinogen in an Ouchterlony plate, and by immunoelectrophoresis, it had a slightly faster than normal anodal migration. The patient’s plasma and purified fibrinogen were able to prolong the clotting time of normal plasma. Using SDS-polyacrylamide gel electrophoresis, we have detected a marked delay in release of a major proportion of the A peptide and a lesser delay in release of the B peptide, after thrombin treatment. B peptide release appeared to be completed before that of A peptide release, in contrast to the situation with normal fibrinogen. Cross-linking of the resulting fibrin was delayed but eventually complete. The rate and extent of monomer aggregation, measured by spectrophotometry, were within normal limits.

ANTI-THROMBINneo IgG IN AN ASYMPTOMATIC PATIENT WITH A BENIGN MONOCLONAL GAMMOPATHY

1987 ◽  
Author(s):  
B S Coller ◽  
J Jesty
Keyword(s):  
Monoclonal Gammopathy ◽  
Asymptomatic Patient ◽  
Protein A ◽  
Thrombin Time ◽  
X Rays ◽  
Excessive Bleeding ◽  
Bone Marrow Histology ◽  
Benign Monoclonal Gammopathy ◽  
History Of ◽  
And Control

A 68 year old male hospitalized for cardiac disease was found to have an elevated prothrombin time (18.5/12.4 s) and aPTT (59.6/25.9s). He had no history of excessive bleeding or bruising. Subsequent evaluation revealed: thrombin time >500/34.1 s; fibrinogen 260 mg/dl functional and 522 mg/dl immunologic; reptilase 25.6/18.1 s; thrombin-induced platelet release of ATP (patient=0 and control=14.6 nmoles/109 platelets at 0.5 U/ml); AT-III 89% functional and 36.5 mg/dl immunologic; and prothrombin 167%. Mixing experiments showed the presence of an inhibitor of the thrombin time, and purification of IgG by protein A affinity chromatography showed the inhibitor of fibrin formation to reside in the IgG fraction. When coupled to Affi-gel 10, patient IgG (but not control IgG) removed purified thrombin from solution; the same gel did not remove prothrombin. The patient's IgG did not inhibit thrombin’s cleavage of a chromogenic substrate (Chromozym TH). Studies on the patient's serum revealed: IgG 2,360 mg/dl, IgA 371 mg/dl, and IgM 107 mg/dl. Serum protein electrophoresis and immunoelectrophoresis showed a monoclonal IgG lambda protein with probably normal amounts of normal IgG. Other parameters (hematocrit, albumin, calcium, bone marrow histology, bone X-rays) indicated that the patient has a benign monoclonal gammopathy, not multiple myeloma. We conclude that our patient is producing an IgG inhibitor that reacts with a neo-antigen produced by the cleavage of prothrombin to thrombin; the IgG can prevent the interaction of thrombin with fibrinogen and the thrombin receptor on platelets, but not small synthetic substrates. We suspect that his monoclonal IgG is the inhibitor and find it remarkable that he has no increase in bleeding.

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.

A Novel Dysfibrinogenemia with Abnormal γ-Chain(Fibrinogen Nagoya).

1979 ◽  
Author(s):  
Junki Takamatsu ◽  
Kanji Oqata ◽  
Tadashi Kamiya ◽  
Katsuo Koie ◽  
Takagi Takashi ◽  
...  
Keyword(s):  
Amino Acid ◽  
Inhibitory Effect ◽  
Chain Structure ◽  
Japanese Family ◽  
Electrophoretic Mobilities ◽  
Sds Page ◽  
History Of ◽  
The Difference ◽  
Fibrin Monomers ◽  
Major Defect

Six individuals in 3 generations of Japanese family had prolonged thrombin clotting time, but no history of hemorrhagic or thrombotic disease. Very low fibrinogen levels were obtained by thrombin clottable protein, while immunological procedures gave normal values of fibrinogen. The serum contained 40-80μg/ml of unclottable fibrinogen related antigens. The patients’ plasma had an inhibitory effect on the fibrin formation in normal plasma. Major defect of this fibrinogen was a delayed aggregation of fibrin monomers.On CM-chromatography (CM-52) of the S-carboxymethylated fibrinogen, three different γ-chains, named γx, γL and γR, were separated. They did not differ in their electrophoretic mobilities in SDS-PAGE, but were distinguishable in PAGE containing 8M urea. Moreover, the amino acid compositions and tryptic peptide mappings of each chain revealed a little difference from those of normal fibrinogen γ chains, suggesting the difference in amino acid substitution or oligosaccharide chain structure.Based on these findings, we designated this fibrinogen as fibrinogen Nagoya; its possible identity without other dysfibrinogenemia has not been excluded.

Autoimmune Antibody in a Hemorrhagic Patient Interacts With Thrombin-Activated Factor XIII in a Unique Manner

Blood ◽  
1999 ◽  
Vol 93 (3) ◽  
pp. 909-917 ◽  
Author(s):  
Laszlo Lorand ◽  
Pauline T. Velasco ◽  
S.N. Prasanna Murthy ◽  
Phil Lefebvre ◽  
David Green
Keyword(s):  
Factor Xiii ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Prior History ◽  
Hemorrhagic Disease ◽  
Normal Range ◽  
Main Target ◽  
History Of ◽  
Unique Manner ◽  
Range Of Values

Abstract Without a prior history of hemorrhagic disease, a 62-year-old man suffered recurrent episodes of bleeding. Solubility of the patient’s clot in 5 mol/L urea indicated a problem with fibrin stabilization. The transamidase activity potential of factor XIII, measured by the incorporation of radioactive putrescine into N,N-dimethylcasein as test substrate, was 62% of control, close to the normal range of values. Examination of the patient’s clot from recalcified plasma by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that essentially none of the  chains and only about two thirds of the γ chains of fibrin became cross-linked under conditions where both were fully cross-linked in the controls. An antibody to factor XIII was isolated which, although recognizing the recombinant rA2subunits, as well as the virgin A2B2 plasma ensemble, showed a 100-fold greater affinity for the thrombin-activated rA2′ and A2′B2 forms of the zymogen, suggesting that the latter would be its main target during coagulation. Furthermore, the patient’s IgG has an ability, never seen before, for inducing an enzymatically active configuration in the thrombin-activated zymogen in the absence of Ca2+.

Fibrinogen Milano XII: a dysfunctional variant containing 2 amino acid substitutions, Aα R16C and γ G165R

Blood ◽  
2001 ◽  
Vol 98 (2) ◽  
pp. 351-357 ◽  
Author(s):  
Bettina Bolliger-Stucki ◽  
Susan T. Lord ◽  
Miha Furlan
Keyword(s):  
Dna Analysis ◽  
Ethylenediaminetetraacetic Acid ◽  
Degradation Products ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Structural Defects ◽  
Thrombin Time ◽  
Sds Page ◽  
Α Helix ◽  
Immunologic Assays

Fibrinogen Milano XII was detected in an asymptomatic Italian woman, whose routine coagulation test results revealed a prolonged thrombin time. Fibrinogen levels in functional assays were considerably lower than levels in immunologic assays. Polymerization of purified fibrinogen was strongly impaired in the presence of calcium or ethylenediaminetetraacetic acid (EDTA). Two heterozygous structural defects were detected by DNA analysis: Aα R16C and γ G165R. As seen previously with other heterozygous Aα R16C variants, thrombin-catalyzed release of fibrinopeptide A was 50% of normal. Additionally, the release of fibrinopeptide B was delayed. Immunoblotting analysis with antibodies to human serum albumin indicated that albumin is bound to Aα 16 C. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) analysis of plasmin digests of fibrinogen Milano XII in the presence of calcium or EDTA showed both normal and novel D1 and D3 fragments. Further digestion of abnormal D3 fragments by chymotrypsin resulted in degradation products of the same size as the fragments derived from normal fibrinogen. SDS-PAGE analysis under reducing conditions showed no difference between normal fibrinogen and fibrinogen Milano XII or between their plasmic fragments. Circular dichroism analysis revealed a shift in the mean residual ellipticity and a significant reduction of the α-helix content in the variant D3 fragment. It is concluded that the Aα-chain substitution is mainly responsible for the coagulation abnormalities, whereas the substitution in the γ-chain induced a conformational change in the D3 fragment.

scholarly journals An apparently higher molecular weight gamma-chain variant in a new congenital abnormal fibrinogen Tochigi characterized by the replacement of gamma arginine-275 by cysteine

Blood ◽  
1988 ◽  
Vol 71 (2) ◽  
pp. 480-487
Author(s):  
N Yoshida ◽  
K Ota ◽  
M Moroi ◽  
M Matsuda
Keyword(s):  
Molecular Weight ◽  
High Performance ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Positive Staining ◽  
Gamma Chain ◽  
Peptide Peak ◽  
Sds Page ◽  
Fibrin Monomers ◽  
Chain Variant

A gamma-chain variant with an apparently higher molecular weight than the normal gamma-chain was detected in a new congenital abnormal fibrinogen with impaired polymerization of the fibrin monomer and with normal release of fibrinopeptides A and B in a 51-year-old male. Purified fibrinogen analyzed on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under the reduced condition in the system of Laemmli contained two protein bands in the gamma-chain region (molecular weight, 50,500 as compared with 50,000 for the normal), both with normal crosslinking ability. The presence of two types of gamma- chains was more clearly detected when reduced and carboxymethylated fibrinogen was analyzed by SDS-PAGE or when reduced fragment D2 was analyzed on SDS-PAGE followed by Western blotting, and identified by positive staining for anti gamma-chain monoclonal antibody. Cyanogen bromide- or lysylendopeptidase-cleavage of purified gamma-chains analyzed on reverse-phase high performance liquid chromatography showed the decrease of one peptide compared with the normal and the appearance of an abnormal peptide peak. Amino acid sequence analysis demonstrated that the gamma arginine-275 of gamma-chain variant was replaced by a cysteine. These data suggest that some regions or conformations containing gamma 275 will affect the polymerization of fibrin monomers. The propositus' two daughters had the same abnormal fibrinogen. This unique inherited abnormal fibrinogen was designated as fibrinogen Tochigi, and the gamma-chain variant as gamma Tochigi.

scholarly journals Fibrinogen Kyoto II, a new congenitally abnormal molecule, characterized by the replacement of A alpha proline-18 by leucine

Blood ◽  
1991 ◽  
Vol 78 (1) ◽  
pp. 149-153
Author(s):  
N Yoshida ◽  
M Okuma ◽  
H Hirata ◽  
M Matsuda ◽  
K Yamazumi ◽  
...  
Keyword(s):  
Amino Acid ◽  
High Performance ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Amino Acid Replacement ◽  
Amino Acid Sequence Analysis ◽  
Alpha Chain ◽  
Molecular Weights ◽  
Peptide Peak ◽  
Fibrin Monomers

A new case of heterozygous dysfibrinogenemia characterized by an amino acid replacement in the NH2-terminal region of the fibrin alpha-chain was found in a 27-year-old woman with a bleeding problem. Her one-stage prothrombin time and activated partial thromboplastin time were slightly prolonged, and the purified fibrinogen from this patient had a markedly prolonged thrombin or reptilase time. Release of fibrinopeptides A and B was normal, but the polymerization of fibrin monomers was impaired. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified fibrinogen under the reduced condition showed no abnormalities in the apparent molecular weights of its three chains. Reverse-phase high performance liquid chromatography (HPLC) of the lysylendopeptidase-cleaved purified A alpha-chains showed a decrease in one peptide compared with the normal amount and the appearance of an abnormal peptide peak. These peptides were treated with thrombin and further separated on HPLC. Amino acid sequence analysis of the abnormal peptide indicated that A alpha proline-18, the second residue from the NH2-terminus of the fibrin alpha-chain, was replaced by leucine. The synthetic peptide Gly-Pro-Arg-Pro inhibited both thrombin- and reptilase-induced fibrin aggregation, but Gly-Leu- Arg-Pro showed little or no inhibition under the same conditions. The discovery of this abnormal fibrinogen supports the findings that A alpha proline-18 is important as part of the polymerization site in the NH2-terminus of the fibrin alpha-chain. The propositus' mother had the same abnormal fibrinogen. This unique inherited abnormal fibrinogen was designated as fibrinogen Kyoto II.

scholarly journals Abnormal adherence-related functions of neutrophils, monocytes, and Epstein-Barr virus-transformed B cells in a patient with C3bi receptor deficiency

Blood ◽  
1985 ◽  
Vol 65 (6) ◽  
pp. 1382-1390 ◽  
Author(s):  
ES Buescher ◽  
T Gaither ◽  
J Nath ◽  
JI Gallin
Keyword(s):  
B Cells ◽  
Epstein Barr Virus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Oxygen Metabolism ◽  
Barr Virus ◽  
History Of ◽  
Severe Periodontal Disease ◽  
Epstein Barr ◽  
Granule Secretion

Abstract We evaluated a 3-year-old female patient with leukocytosis, recurrent infections, severe periodontal disease, and a history of delayed separation of the umbilical stump. This patient's polymorphonuclear leukocytes (PMNs) had normal membrane depolarization responses, normal oxygen metabolism, normal granule secretion responses, normal bactericidal activity, and normal C3b rosetting. However, by fluorescent cell analysis and C3bi rosetting, it was determined that her cells lacked the C3bi receptor. In addition, the patient's PMNs showed markedly abnormal chemotaxis, adherence, and aggregation responses, and partial abnormalities were detected in PMN spreading and phagocytosis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the subject's neutrophil cytoplasts were missing a 180,000-dalton moiety. Her monocytes also had defective chemotaxis and failed to adhere and grow normally in culture. Epstein-Barr virus- transformed B cells from the patient lacked an aggregation response to phorbol myristate acetate. Laboratory and clinical evaluations of this patient's mother showed no abnormalities. These studies demonstrate that C3bi receptor deficiency can be associated with functional abnormalities in multiple myeloid cells and that the absence of C3bi receptor is associated with abnormal adherence-related functions of these cells.

scholarly journals Fibrinogen Kyoto II, a new congenitally abnormal molecule, characterized by the replacement of A alpha proline-18 by leucine

Blood ◽  
1991 ◽  
Vol 78 (1) ◽  
pp. 149-153 ◽  
Author(s):  
N Yoshida ◽  
M Okuma ◽  
H Hirata ◽  
M Matsuda ◽  
K Yamazumi ◽  
...  
Keyword(s):  
Amino Acid ◽  
High Performance ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Amino Acid Replacement ◽  
Amino Acid Sequence Analysis ◽  
Alpha Chain ◽  
Molecular Weights ◽  
Peptide Peak ◽  
Fibrin Monomers

Abstract A new case of heterozygous dysfibrinogenemia characterized by an amino acid replacement in the NH2-terminal region of the fibrin alpha-chain was found in a 27-year-old woman with a bleeding problem. Her one-stage prothrombin time and activated partial thromboplastin time were slightly prolonged, and the purified fibrinogen from this patient had a markedly prolonged thrombin or reptilase time. Release of fibrinopeptides A and B was normal, but the polymerization of fibrin monomers was impaired. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified fibrinogen under the reduced condition showed no abnormalities in the apparent molecular weights of its three chains. Reverse-phase high performance liquid chromatography (HPLC) of the lysylendopeptidase-cleaved purified A alpha-chains showed a decrease in one peptide compared with the normal amount and the appearance of an abnormal peptide peak. These peptides were treated with thrombin and further separated on HPLC. Amino acid sequence analysis of the abnormal peptide indicated that A alpha proline-18, the second residue from the NH2-terminus of the fibrin alpha-chain, was replaced by leucine. The synthetic peptide Gly-Pro-Arg-Pro inhibited both thrombin- and reptilase-induced fibrin aggregation, but Gly-Leu- Arg-Pro showed little or no inhibition under the same conditions. The discovery of this abnormal fibrinogen supports the findings that A alpha proline-18 is important as part of the polymerization site in the NH2-terminus of the fibrin alpha-chain. The propositus' mother had the same abnormal fibrinogen. This unique inherited abnormal fibrinogen was designated as fibrinogen Kyoto II.

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