Automated amidolytic method for determining heparin, a heparinoid, and a low-Mr heparin fragment, based on their anti-Xa activity.

Cate, Hugo Ten; Lamping, R. J.; Henny, C. P.; Prins, Astrid H.; Cate, J. W. Ten

doi:10.1093/clinchem/30.6.860

Cited by 41 publications

(8 citation statements)

References 10 publications

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“…Thrombin‐AT complexes (TATc; Dade Behring, Marburg, Germany) and D‐dimer (Diagnostica Stago, Roche, Almere, the Netherlands) were measured by ELISA. AT [16] and plasminogen activator (PA) activity [17] were measured by automated amidolytic techniques. Tissue‐type plasminogen activator (t‐PA) antigen was measured with an ELISA.…”

Section: Methodsmentioning

confidence: 99%

Vagus nerve stimulation inhibits activation of coagulation and fibrinolysis during endotoxemia in rats

Westerloo

Giebelen

Meijers

et al. 2006

Journal of Thrombosis and Haemostasis

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To cite this article: van Westerloo DJ, Giebelen IAJ, Meijers JCM, Daalhuisen J, de Vos AF, Levi M, van der Poll T. Vagus nerve stimulation inhibits activation of coagulation and fibrinolysis during endotoxemia in rats. J Thromb Haemost 2006; 4: 1997-2002 Summary. Background: Sepsis and endotoxemia are associated with concurrent activation of inflammation and the hemostatic mechanism, which both contribute to organ dysfunction and death. Electrical vagus nerve stimulation (VNS) has been found to inhibit tumor necrosis factor (TNF)-a release during endotoxemia in rodents. Objective: To determine the effect of VNS on activation of coagulation and fibrinolysis. Methods: Rats received a sublethal i.v. dose of lipopolysaccharide (LPS) after electrical VNS or sham stimulation. Activation of coagulation and fibrinolysis, as well as cytokine release, was measured before LPS injection and 2, 4 and 6 h thereafter. Results: LPS induced activation of the coagulation system (increases in the plasma concentrations of thrombin-antithrombin complexes and D-dimer, and a decrease in antithrombin) and biphasic changes in the fibrinolytic system [early rises of plasminogen activator activity and tissue-type plasminogen activator, followed by a delayed increase in plasminogen activator inhibitor type 1 (PAI-1)]. VNS strongly inhibited all LPS-induced procoagulant responses and more modestly attenuated the fibrinolytic response. In addition, VNS attenuated the LPS-induced increases in plasma and splenic concentrations of the proinflammatory cytokines TNF-a and interleukin-6 (IL-6), while not influencing the release of the anti-inflammatory cytokine IL-10. Conclusion: These data illustrate a thus far unrecognized effect of VNS and suggest that the cholinergic anti-inflammatory pathway not only impacts on inflammation but also on the coagulantanticoagulant balance.

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Section: Methodsmentioning

confidence: 99%

Vagus nerve stimulation inhibits activation of coagulation and fibrinolysis during endotoxemia in rats

Westerloo

Giebelen

Meijers

et al. 2006

Journal of Thrombosis and Haemostasis

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“…It has been over 90 years since the discovery of heparin as an effective anticoagulant. The mechanism of anticoagulant is to activate antithrombin III (ATIII) by the allosteric effect through the speci c combination of polysaccharide sequence and ATIII [12][13][14] . The activated ATIII can inhibit a variety of coagulation factors including Xa, IIa, IXa, XIa, XIIa etc., to exert the anticoagulant activity.…”

Section: Discussionmentioning

confidence: 99%

Release Characteristics of Enoxaparin Sodium Loaded Polymethylmethacrylate Bone Cement

Sun

et al. 2020

Preprint

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Background: This study aimed to prepare the polymethylmethacrylate (PMMA) bone cement release system with different concentrations of enoxaparin sodium (ES) and to investigate the release characteristics of ES after loading into the PMMA bone cement. Methods: In the experimental group, 40g Palacos®R PMMA bone cement was loaded with various amount of ES 4000, 8000, 12000, 16000, 20000, and 24000 AXaIU, respectively. The control group was not loaded with ES. Scanning electron microscopy (SEM) was used to observe the surface microstructure of the bone cement in the two groups. In the experiment group, the mold was extracted continuously with pH7.4 Tris-HCL buffer for 10 days. The extract solution was collected every day and the anti-FXa potency was measured. The experiment design and statistical analysis were conducted using a quantitative response parallel line method. Results: Under the SEM, it was observed that ES was filled in the pores of PMMA bone cement polymer structure and released from the pores after extraction. There was a burst effect of the release. The release amount of ES on the first day was 0.415, 0.858, 1.110, 1.564, 1.952 and 2.513 respectively from the six groups with various ES loading amount of 4000, 8000, 12000, 16000, 20000 and 24000 AXaIU, all reaching the peak of release on the first day. The release decreased rapidly on the next day and entered the plateau phase on the fourth day. Conclusion: PMMA bone cement can be used as a carrier to effectively release enoxaparin sodium within a short term.

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“…Determination of the activated partial thromboplastin time (aPTT) was performed according to unfractionated procedures in citrated plasma. The chromogenic anti-factor Xa activity assay was performed using an EPOS 5060 analyzer, as described previously (20), and calibrated against a Fraxiparin standard. The plasma anti-factor Ila assay was performed as described elsewhere (21 ).…”

Section: Blood Sampling and Assaysmentioning

confidence: 99%

Additive Effect of the Combined Administration of Low Molecular Weight Heparin and Recombinant Hirudin on Thrombus Growth in a Rabbit Jugular Vein Thrombosis Model

et al. 1994

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SummaryRecombinant Hirudin (r-Hirudin) is a new anticoagulant with specific antithrombin activity independently of antithrombin III. Low molecular weight heparins (LMWH) exert predominantly anti-Xa activity. Therefore, we hypothesized that combined administration of r-Hirudin and LMWH would induce a stronger antithrombotic effect as compared to r-Hirudin administered alone or combined with unfractionated heparin. To assess the effect on thrombus growth, we determined the accretion of l25I-labeled fibrinogen onto autologous non-radioactive thrombi preformed in the jugular veins of rabbits. The rabbits received unfractionated heparin (80 anti-factor Xa U), LMWH (80 anti-factor Xa U) or r-Hirudin (0.3, 5.0 and 10.0 mg/kg) either separately or by combined infusion for a 3 h period.R-Hirudin reduced the thrombus growth in a dose dependent fashion. The combined administration of 80 anti-Xa U LMWH and r-Hirudin at a dose of 0.3 mg/kg resulted in a stronger antithrombotic effect as compared to the combined infusion of unfractionated heparin and r-Hirudin (thrombus growth: 14.3% ± 6.0 vs 28.9% ± 6.5; p = 0.001). This difference in additive antithrombotic effect of 80 anti-Xa ULMWH versus unfractionated heparin on r-Hirudin was also observed when LMWH was combined with 5.0 mg/kg and 10.0 mg/kg r-Hirudin versus unfractionated heparin combined with r-Hirudin (thrombus growth: 16.4% ± 1.6 vs 29.1% ± 3.9; p = 0.01 and 10.1% ± 1.8 vs 20.4% ± 4.5; p = 0.001, respectively).In conclusion, this study showed an additive antithrombotic effect of LMWH on the thrombus growth reducing effect of r-Hirudin. This may potentially be of clinical interest, i. e. much lower doses of r-Hirudin may be used to achieve a similar effective anticoagulant effect when combined with LMWH.

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Automated amidolytic method for determining heparin, a heparinoid, and a low-Mr heparin fragment, based on their anti-Xa activity.

Cited by 41 publications

References 10 publications

Vagus nerve stimulation inhibits activation of coagulation and fibrinolysis during endotoxemia in rats

Vagus nerve stimulation inhibits activation of coagulation and fibrinolysis during endotoxemia in rats

Release Characteristics of Enoxaparin Sodium Loaded Polymethylmethacrylate Bone Cement

Additive Effect of the Combined Administration of Low Molecular Weight Heparin and Recombinant Hirudin on Thrombus Growth in a Rabbit Jugular Vein Thrombosis Model

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