Reproducibility and temporal stability of ADP‐induced platelet aggregation: Comparison of the anticoagulants sodium citrate and D‐phenylalanyl‐L‐prolyl‐L‐arginyl chloromethyl ketone

Horne, William C.; Cohen, Donald J.; Anderson, George M.

doi:10.1002/ajh.2830380108

Cited by 4 publications

(2 citation statements)

References 27 publications

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“…This agrees with findings by Horne et al. [14], who found a more reproducible and stable ADPTPA in PRP anticoagulated with PPACK compared with citrate obtained from nine individuals and stored between 30 min and 4.5 h after venipuncture.…”

Section: Discussionsupporting

confidence: 93%

“…Previous studies have established specific direct thrombin inhibitors, such as hirudin or PPACK, as anticoagulants for platelet function studies because these agents, unlike citrate, maintain physiological levels of calcium ions and have no direct effect on platelets [5,13–16]. However, the different effects of citrate and direct thrombin inhibitors on platelet storage lesion have not been examined systematically over longer times after blood collection.…”

Section: Discussionmentioning

confidence: 99%

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Preservation of in vitro function of platelets stored in the presence of a synthetic dual inhibitor of factor Xa and thrombin

Hellstern¹,

Stürzebecher

Wuchold

et al. 2007

Journal of Thrombosis and Haemostasis

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Summary. Background: The use of citrate anticoagulant limits the clinical significance of platelet function tests. Thrombin inhibitors cannot prevent thrombin-induced platelet activation completely. We examined the influence of benzylsulfonyl-D-Arg-Pro-4-amidinobenzylamide (BAPA), a dual inhibitor of Factor Xa (FXa) and thrombin, on platelet responsiveness to agonists when measured between 2 and 24 h after venipuncture. Methods:Blood samples from 36 individuals were anticoagulated with citrate and BAPA, respectively. Turbidimetric platelet aggregometry (TPA) and impedance platelet aggregometry (IPA), a whole blood platelet counting assay for measuring platelet aggregation (PCA), and Platelet Function Anlayzer-100 TM (PFA-100 TM ) closure times (CTs) were determined after whole blood storage between 2 and 24 h after venipuncture. Native whole blood was studied over 48 h to determine the inhibition of thrombin generation by BAPA, hirudin and melagatran. Results: BAPA inhibited thrombin generation completely for 48 h, while hirudin and melagatran did not. The use of citrate resulted in significantly reduced TPA induced by arachidonic acid (AA) or adenosine 5¢-diphosphate (ADP), and significantly reduced IPA regardless of agonist when measured 10 and 24 h after blood collection. PCA ratios in citrated blood also dropped significantly 10 and 24 h after venipuncture. The length of storage of BAPAanticoagulated blood samples over 24 h had no significant influence on any platelet response. The reproducibility of platelet function assay results obtained from BAPA-anticoagulated samples was significantly better than corresponding data from citrated blood. Conclusion: TPA, IPA, PCA or PFA-100 TM CTs remain stable for 24 h when whole blood is anticoagulated with a dual inhibitor of FXa and thrombin. This would greatly simplify the shipment of samples for platelet function testing.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Preservation of in vitro function of platelets stored in the presence of a synthetic dual inhibitor of factor Xa and thrombin

Hellstern¹,

Stürzebecher

Wuchold

et al. 2007

Journal of Thrombosis and Haemostasis

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Heterogeneity of the aggregation response of human platelets to arginine vasopressin

et al. 1995

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Previous reports have alluded to variability in the aggregation response of normal human platelets to the neuropeptide arginine vasopressin (AVP). Since it has not been well documented, the current studies were undertaken to characterize this response. AVP (1-100 nM) produced a concentration-dependent aggregation response. Although the aggregation response to 100 nM AVP did not correlate with age or sex, there was a bimodal response distribution based on the presence or absence of a second wave of aggregation. In kinetic studies, the apparent km of AVP was 18.3 +/- 5.4 nM. There was a significant inverse relationship between the maximal aggregation response to 100 nM AVP and the km (r = -0.82). One hundred nanomolar AVP increased the intracellular calcium concentration of platelets by 406 +/- 120 nM in calcium free buffer and by 658 +/- 233 nM in the presence of 1.0 mM CaCl2. The aggregation response to 100 nM AVP correlated most strongly with the transmembrane influx of calcium (r = 0.84). In individuals whom 100 nM AVP was able to generate a second wave of aggregation, the selective protein kinase C inhibitor bis-indolylmaleimide significantly decreased the platelet aggregation response. Thus, there is significant heterogeneity in the aggregation response of normal human platelets to AVP. Based on our kinetic studies and the effects of PKC inhibition on the aggregation response to AVP, we would hypothesize that the variability of the aggregation response of normal human platelets to AVP is related to a polymorphism of the platelet AVP V1 receptor.

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Use of thrombin inhibitors ex vivo allows critical care clinical chemistry and hematology testing on common specimens

Lyon

Harding

Drobot

et al. 1997

Clinical Biochemistry

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Reproducibility and temporal stability of ADP‐induced platelet aggregation: Comparison of the anticoagulants sodium citrate and D‐phenylalanyl‐L‐prolyl‐L‐arginyl chloromethyl ketone

Cited by 4 publications

References 27 publications

Preservation of in vitro function of platelets stored in the presence of a synthetic dual inhibitor of factor Xa and thrombin

Preservation of in vitro function of platelets stored in the presence of a synthetic dual inhibitor of factor Xa and thrombin

Heterogeneity of the aggregation response of human platelets to arginine vasopressin

Use of thrombin inhibitors ex vivo allows critical care clinical chemistry and hematology testing on common specimens

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