PubMed HealthSearch

PubMed · 6996867

New perspectives in coagulation testing.

Abstract

The field of coagulation testing has undergone some major technological and conceptual developments, which are briefly reviewed here. The assessment of coagulation parameters is no longer restricted to the study of clot formation and its dissolution. The understanding of the biochemical nature of coagulation processes, coupled with the development of new therapeutic agents in the treatment of hemostatic disorders, has brought about the development of fast, reliable, and clearly defined laboratory test procedures to evaluate the components of this system. The introduction of methods involving synthetic substrates has been very significant because many of the coagulation parameters can now be measured with a spectrophotometer or fluorometer, by methods that lend themselves to the automation found in most large clinical chemistry laboratories. In our laboratory, we use automated synthetic-substrate methods for antithrombin-III, plasminogen, and prothrombin, and are developing the synthetic-substrate assay equivalent of clot-based prothrombin time and partial thromboplastin. Immunological methods such as laser/rate nephelometry, enzyme-linked immunoassays, electroimmunodiffusion, and radioimmunoassays have been utilized to evaluate coagulation proteins. The relation of functional and immunological properties of these proteins to their physiological function is being studied. In coming years the testing of coagulation function will undergo some major changes and will require input from clinical chemists and other laboratory scientists to facilitate the technology transfer and proper standardization of new methods.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

J Fareed, H L Messmore, E W Bermes. 1980. New perspectives in coagulation testing.. https://pubmed.ncbi.nlm.nih.gov/6996867/

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

Fluorescence and circular dichroism studies during the interactions of sulfated polysaccharides with antithrombin III.

The changes in relative fluorescence of antithrombin III (AT-III) during its interaction with sulfated xylans were compared with that of sulfated glycosaminoglycans by measuring the ratio of the increase in fluorescence of AT-III in the presence of sulfated polysaccharide to the fluorescence of AT-III alone for various mass ratios. Interactions of corn cob xylan sulfate (CCXS) and sodium pentosan polysulfate (SP-54) with AT-III resulted in enhancements of relative fluorescence which were lower than commercial heparin. At mass ratios below 1, heparan sulfate and low molecular weight heparin (LMWH) gave increases in the relative fluorescence higher than that of commercial heparin, while highly sulfated semisynthetic chondroitin sulfates A and C gave much smaller increases. The relative fluorescence enhancements of AT-III by heparan sulfate, commercial heparin, LMWH and heparin derived pentasaccharide (HDP) increased with increasing mass ratios while the enhancements by CCXS, SP-54 and the highly sulfated chondroitin sulfates A and C were reversed at higher mass ratios. The estimated dissociation constants (kd) for the interaction of AT-III and the heparin-related compounds showed that heparin sulfate and LMWH gave the lowest kd values indicating a higher affinity for AT-III while commercial heparin and HDP gave higher kd values, indicating a lower affinity for AT-III. SP-54 gave a kd value lower than CCXS, indicating a greater affinity for AT-III. A comparison of the near ultraviolet (UV) circular dichroism (CD) spectrum of AT-III alone and during its interaction with oat spelts xylan sulfate (OSXS) showed enhancements of the two aromatic amino acid regions corresponding to phenylalanine and tryptophan.

Antithrombin III

Inactivation of factor XIa in human plasma assessed by measuring factor XIa-protease inhibitor complexes: major role for C1-inhibitor.

From experiments with purified proteins, it has been concluded that factor XIa (FXIa) is inhibited in plasma mainly by alpha 1-antitrypsin (a1AT), followed by antithrombin III (ATIII), C1-inhibitor (C1Inh), and alpha 2-antiplasmin (a2AP). However, the validity of this concept has never been studied in plasma. We established the relative contribution of different inhibitors to the inactivation of FXIa in human plasma, using enzyme-linked immunosorbent assays (ELISAs) for the quantification of complexes of FXIa with a1AT, C1Inh, a2AP, and ATIII. We found that 47% of FXIa added to plasma formed complexes with C1Inh, 24.5% with a2AP, 23.5% with a1AT, and 5% with ATIII. The distribution of FXIa between these inhibitors in plasma was independent of whether FXIa was added to plasma, or was activated endogenously by kaolin, celite, or glass. However, in the presence of heparin (1 or 50 U/mL), C1Inh appeared to be the major inhibitor of FXIa, followed by ATIII. Furthermore, at lower temperatures, less FXIa-C1Inh and FXIa-a1AT complexes but more FXIa-a2AP complexes were formed. These data demonstrate that the contribution of the different inhibitors to inactivation of FXIa in plasma may vary, but C1Inh is the principal inhibitor under most conditions.

Antithrombin III

Mechanism of antithrombin III inhibition of factor VIIa/tissue factor activity on cell surfaces. Comparison with tissue factor pathway inhibitor/factor Xa-induced inhibition of factor VIIa/tissue factor activity.

Recent studies have shown that antithrombin III (AT III)/heparin is capable of inhibiting the catalytic activity of factor VIIa bound either to relipidated tissue factor (TF) in suspension or to TF expressed on cell surfaces. We report studies of the mechanism of which by AT III inhibits factor VIIa bound to cell surface TF and compare this inhibitory mechanism with that of tissue factor pathway inhibitor (TFPI)-induced inhibition of factor VIIa/TF. AT III alone and AT III/heparin to a greater extent reduced factor VIIa bound to cell surface TF. Our data show that the decrease in the amount of factor VIIa associated with cell surface TF in the presence of AT III was the result of (1) accelerated dissociation of factor VIIa from cell surface TF after the binding of AT III to factor VIIa/TF complexes and (2) the inability of the resultant free factor VIIa-AT III complexes to bind effectively to a new cell surface TF site. Binding of TFPI/factor Xa to cell surface factor VIIa/TF complexes markedly decreased the dissociation of factor VIIa from the resultant quaternary complex of factor VIIa/TF/TFPI/factor Xa. Addition of high concentrations of factor VIIa could reverse the AT III-induced inhibition of cell surface factor VIIa/TF activity but not TFPI/factor Xa-induced inhibition of factor VIIa/TF activity.

Antithrombin III