Assessment of complex peptide degradation pathways via structured multicompartmental modeling approaches: The metabolism of dynorphin A1–13 and related fragments in human plasma

SummaryA mixture of adsorbed normal human plasma and chicken plasma was prepared as reagent for factor IX measurement using a one-stage method. The substrate was found to be specific for factor IX. Its performances tested on samples displaying factor IX activity ranging from <l%–2,500% compared favorably with those obtained when using the plasma of severe haemophilia B patients as substrate.

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Plasma Components which Interfere with Ristocetin-induced Platelet Aggregation

Thrombosis and Haemostasis ◽

10.1055/s-0038-1647847 ◽

1975 ◽

Vol 33 (03) ◽

pp. 540-546 ◽

Cited By ~ 3

Author(s):

Robert F Baugh ◽

James E Brown ◽

Cecil Hougie

Keyword(s):

Platelet Aggregation ◽

Human Plasma ◽

Plasma Proteins ◽

Binding Protein ◽

Plasma Heating ◽

Protein S ◽

Fold Increase ◽

Normal Human Plasma ◽

Preliminary Examination ◽

Normal Human

SummaryNormal human plasma contains a component or components which interfere with ristocetin-induced platelet aggregation. Preliminary examination suggests a protein (or proteins) which binds ristocetin and competes more effectively for ristocetin than do the proteins involved in ristocetin-induced platelet aggregation. The presence of this protein in normal human plasma also prevents ristocetin-induced precipitation of plasma proteins at levels of ristocetin necessary to produce platelet aggregation (0.5–2.0 mg/ml). Serum contains an apparent two-fold increase of this component when compared with plasma. Heating serum at 56° for one hour results in an additional 2 to 4 fold increase. The presence of a ristocetin-binding protein in normal human plasma requires that this protein be saturated with ristocetin before ristocetin-induced platelet aggregation will occur. Variations in the ristocetin-binding protein(s) will cause apparent discrepancies in ristocetin-induced platelet aggregation in normal human plasmas.

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Studies on the Fibrinolytic System in Human Plasma: Quantitative Determination of Plasminogen Activators and Proactivators

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646786 ◽

1979 ◽

Vol 41 (02) ◽

pp. 365-383 ◽

Cited By ~ 80

Author(s):

C Kluft

Keyword(s):

Pilot Study ◽

Plasma Level ◽

Human Plasma ◽

Plasminogen Activators ◽

Venous Occlusion ◽

Quantitative Information ◽

Optimal Recovery ◽

Dextran Sulphate ◽

Baseline Levels

SummaryEffects due to plasma plasminogen activators and proactivators are usually studied in assay systems where inhibitors influence the activity and where the degree of activation of proactivators is unknown. Quantitative information on activator and proactivator levels in plasma is therefore not availableStudies on the precipitating and activating properties of dextran sulphate in euglobulin fractionation presented in this paper resulted in the preparation of a fraction in which there was optimal recovery and optimal activation of a number of plasminogen activators and proactivators from human plasma. The quantitative assay of these activators on plasminogen-rich fibrin plates required the addition of flufenamate to eliminate inhibitors. The response on the fibrin plates (lysed zones) could be coverted to arbitrary blood activator units (BAU). Consequently, a new activator assay which enables one to quantitatively determine the plasma level of plasminogen activators and proactivators together is introduced.Two different contributions could be distinguished: an activity originating from extrinsic activator and one originating from intrinsic proactivators. The former could be assayed separately by means of its resistance to inhibition by Cl-inactivator. Considering the relative concentrations of extrinsic and intrinsic activators, an impression of the pattern of activator content in plasma was gained. In morning plasma with baseline levels of fibrinolysis, the amount of extrinsic activator was negligible as compared to the level of potentially active intrinsic activators. Consequently, the new assay nearly exclusively determines the level of intrinsic activators in morning plasma. A pilot study gave a fairly stable level of 100 ± 15 BAU/ml (n = 50). When fibrinolysis was stimulated by venous occlusion (15 min), the amount of extrinsic activator was greatly increased, reaching a total activator level of 249 ± 27 BAU/ml (n = 7).

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Potentially Thrombogenic Materials in Factor IX Concentrates

Thrombosis and Haemostasis ◽

10.1055/s-0038-1647857 ◽

1975 ◽

Vol 33 (03) ◽

pp. 617-631 ◽

Cited By ~ 47

Author(s):

H. S Kingdon ◽

R. L Lundblad ◽

J. J Veltkamp ◽

D. L Aronson

Keyword(s):

Human Plasma ◽

Antithrombin Iii ◽

Factor Ix ◽

In Vitro Assays ◽

Substantial Agreement ◽

Coefficient Of Correlation

SummaryFactor IX concentrates manufactured from human plasma and intended for therapeutic infusion in man have been suspected for some time of being potentially thrombogenic. In the current studies, assays were carried out in vitro and in vivo for potentially thrombogenic materials. It was possible to rank the various materials tested according to the amount of thrombogenic material detected. For concentrates not containing heparin, there was substantial agreement between the in vivo and in vitro assays, with a coefficient of correlation of 0.77. There was no correlation between the assays for thrombogenicity and the antithrombin III content. We conclude that many presently available concentrates of Factor IX contain substantial amounts of potentially thrombogenic enzymes, and that this fact must be considered in arriving at the decision whether or not to use them therapeutically.

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Analysis of Intrinsic Fibrinolysis in Human Plasma Induced by Dextran Sulfate

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648467 ◽

1992 ◽

Vol 67 (04) ◽

pp. 440-444 ◽

Cited By ~ 3

Author(s):

Hiroko Tsuda ◽

Toshiyuki Miyata ◽

Sadaaki Iwanaga ◽

Tetsuro Yamamoto

Keyword(s):

Molecular Weight ◽

Human Plasma ◽

Dextran Sulfate ◽

Tissue Type ◽

Factor Xii ◽

Normal Human Plasma ◽

High Molecular Weight Kininogen ◽

Single Chain ◽

Major Pathway ◽

Normal Human

SummaryThe analysis of normal human plasma by fibrin autography revealed four species of plasminogen activator (PA) activity related to tissue-type PA, factor XII, prekallikrein and urokinase-type PA (u-PA). The u-PA activity increased significantly by incubating plasma with dextran sulfate. This increase was coincident with both the cleavage of factor XII and the complex formation of activated factor XII with its plasma inhibitors, which were determined by immunoblotting procedure. The dextran sulfate-dependent activation of u-PA required both factor XII and prekallikrein, but did not require either plasminogen or factor XI. High molecular weight kininogen was required only at a low concentration of dextran sulfate. Thus the results indicate that the factor XII and prekallikrein-mediated activation of single chain u-PA (scu-PA) operates as a major pathway of scu-PA activation in whole plasma in contact with dextran sulfate.

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Presence and Possible Origin of ɛ(γ-Glutamyl)Lysine Isodipeptide in Human Plasma

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648380 ◽

1992 ◽

Vol 67 (01) ◽

pp. 060-062 ◽

Cited By ~ 9

Author(s):

J Harsfalvi ◽

E Tarcsa ◽

M Udvardy ◽

G Zajka ◽

T Szarvas ◽

...

Keyword(s):

Human Plasma ◽

Fibrinolytic Therapy ◽

Normal Human Plasma ◽

Normal Human ◽

Hplc Technique ◽

Significant Change

Summaryɛ(γ-glutamyl)lysine isodipeptide has been detected in normal human plasma by a sensitive HPLC technique in a concentration of 1.9-3.6 μmol/1. Incubation of in vitro clotted plasma at 37° C for 12 h resulted in an increased amount of isodipeptide, and there was no further significant change when streptokinase was also present. Increased in vivo isodipeptide concentrations were also observed in hypercoagulable states and during fibrinolytic therapy.

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