Effect of the Combination of Mannitol and Ringer Acetate or Hydroxyethyl Starch on Whole Blood Coagulation In Vitro

The changes of factor VIII (F. VIII) during blood coagulation process is still controversial. We analyzed the F. VIII antigen (F. VIII:Ag) at various intervals of in vitro blood clotting by immunoassays using polyclonal and different kinds of monoclonal antibodies to F. VIII.We used two immunoassays, an immunoradiometric assay (IRMA) and an enzyme-linked immunosorbent assay (ELISA). The IRMA was performed by the method of Peake et al. using high-titer allo-antibodies to F. VIII. The ELISA was performed by two-site solid phase system consisting of alloantibodies as the first and one of three kinds of monoclonal antibodies (NMC-VIII/1, -VIII/2 or C5*) as the second antibody. Using the immunoblotting technique, it had been shown that NMC-VIII/1 recognized the 80/79 kDa derived from C-terminus and both NMC-VIII/2 and C5 recognized 54 kDa derived from N-terminus.The mean levels of F. VIII:Ag in 20 normal plasmas and sera were 0.97±0.23 U/ml and 0.68±0.21 U/ml, respectively using the polyclonal IRMA. The mean levels of F. VIII:Ag in normal sera were 0.14±0.05 U/ml (NMC-VIII/1), 0.71±0.21 U/ml (NMC-VIII/2), and 0.012±0.02 U/ml (C5) using the monoclonal ELISAs. In the initial phase of whole blood coagulation in vitro, the increase of F. VIII: Ag was observed by the polyclonal IRMA as F. VIII:C assayed by a one-stage clotting method increased. On the other hand, the F. VIII:Ag assayed by NMC-VIII/1 or C5 monoclonal ELISA progressively decreased to the serum level within 30 min. The F. VIII:Ag by NMC-VIII/2 declined to the serum level at a slow rate. In order to study the influence of thrombin on F. VIIIrAg during blood clotting, a synthesized selective thrombin inhibitor (MD-805, Mitsubishi Chemical Ind.) was previously added to the whole blood tested. The changes of F. VIII:Ag with MD-805 by the monoclonal ELISAs were almost the same as those without MD-805.It is suggested that in the whole blood coagulation process the antigenicity of F. VIII molecule changes in the initial phase (within 30 min.), but that thrombin does not play the main role of the phenomenon in physiological concentration.*C5 was kindly supplied from Dr. C. Fulcher (Scripps Clinic and Research Foundation, USA)

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Role of Serotonin in Blood Coagulation

American Journal of Physiology-Legacy Content ◽

10.1152/ajplegacy.1957.189.3.470 ◽

1957 ◽

Vol 189 (3) ◽

pp. 470-474 ◽

Cited By ~ 8

Author(s):

W. L. Milne ◽

S. H. Cohn

Keyword(s):

Blood Coagulation ◽

Therapeutic Effect ◽

Whole Blood ◽

Active Role ◽

Clotting Time ◽

Platelet Factor ◽

Coagulation Defect ◽

Increased Sensitivity

Effects of serotonin (5HTA) were studied a) in whole blood ( in vitro) from both normal and x-irradiated animal, and b) in isolated clotting systems of plasma and purified fibrinogen. 5HTA was found to play an active role in blood coagulation in addition to its previously demonstrated role as a vasoconstrictor. In whole blood from normal and irradiated animals, and in platelet-deficient plasma, 5HTA acts in a manner similar to platelet factor 2 in accelerating the conversion of fibrinogen to fibrin. In a purified fibrinogen solution, 5HTA inhibits the fibrinogen-fibrin reaction, and thus differs from platelet factor 2. This may be due to the absence of an antithrombin in the purified fibrinogen solution. Serotonin appears to have a therapeutic effect on the postirradiation coagulation defect in that it decreases the prolonged heparin clotting time. This increased sensitivity to heparin during the postirradiation thrombocytopenia may be due to a deficiency of serotonin, which acts as an antagonist to heparin (an antithrombin). The action of 5HTA is postulated as an antagonist of an antithrombin which blocks the fibrinogen to fibrin reaction.

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