title: Determination of the levels of unfractionated and low-molecular-weight heparins in plasma: their effect on thrombin-mediated feedback reactions in vivo. Preliminary results after subcutaneous injection.
abstract: We define a standard independent unit (SIU) of heparin as that amount that, in plasma containing 1 mumol of ATIII, raises the (pseudo-)first-order breakdown constant of factor Xa by 1 min-1. These units measure all material with a high affinity for ATIII (HAM); only material above the critical chain length of 17 monosaccharide units (above critical chain length material; ACLM) catalyzes the inactivation of thrombin. An SIU of ACLM is therefore analogously defined as the amount that, in plasma containing 1 mumol of ATIII, will raise the (pseudo-)first-order breakdown constant of thrombin by 1 min-1. Of any given heparin preparation one can determine the specific HAM and ACLM activities in terms of SIU/mg. On the basis of the factor Xa and thrombin breakdown constants found in a plasma sample one can then determine the levels of HAM and ACLM. Preliminary experiments were carried out in plasma samples obtained after subcutaneous injection of unfractionated heparin (UFH) and of two types of low-molecular-weight heparin (LMWH). About three times more of UFH activity than of LMWH activity has to be injected to obtain the same levels of ACLM in the plasma. Only with the LMWHs significant amounts of BCLM are found, which rises higher and persists longer than the ACLM. We determined the course of thrombin generation in platelet-rich plasma (PRP) and in platelet-poor plasma (PPP), as well as in the PPP factor Xa generation curve and the course of prothrombin conversion. The observed inhibitions correlated much better with the levels of ACLM than with those of below critical chain length material. The difference between UFH and LMWHs can therefore not be explained in terms of antithrombin and anti-factor-Xa activity. The essential difference between UFH and LMWH appears in the feedback effect of thrombin in PRP, where thrombin generation is both inhibited and retarded by LMWH, while it is only retarded but hardly inhibited by UFH.
authors.affiliation: Department of Biochemistry, Rijksuniversiteit Limburg, Maastricht, The Netherlands.
authors.forename: H C
authors.initials: HC
authors.lastname: Hemker
authors.forename: S
authors.initials: S
authors.lastname: Béguin
authors.forename: A V
authors.initials: AV
authors.lastname: Bendetowicz
authors.forename: S
authors.initials: S
authors.lastname: Wielders
mesh_terms: D000328:Adult; D004789:Enzyme Activation; D065427:Factor Xa Inhibitors; D005246:Feedback / Q000187:drug effects; D006495:Heparin, Low-Molecular-Weight / Q000097:blood* / Q000493:pharmacokinetics / Q000494:pharmacology; D006801:Humans; D007279:Injections, Subcutaneous; D008297:Male; D010865:Pilot Projects; D011516:Prothrombin / Q000378:metabolism; D013917:Thrombin / Q000378:metabolism*; D013925:Thromboplastin / Q000378:metabolism; D014894:Weights and Measures
chemical_list: D065427:Factor Xa Inhibitors; D006495:Heparin, Low-Molecular-Weight; D011516:Prothrombin; D013925:Thromboplastin; D013917:Thrombin