Platelet release of β-thromboglobulin and platelet factor 4 and serotonin in plasma samples

Ohkawa, Ryunosuke; Hirowatari, Yuji; Nakamura, Kazuhiro; Ohkubo, Shigeo; Ikeda, Hitoshi; Okada, Mitsumasa; Tozuka, Minoru; Nakahara, Kazuhiko; Yatomi, Yutaka

doi:10.1016/j.clinbiochem.2005.07.008

Cited by 37 publications

(28 citation statements)

References 10 publications

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“…This precaution is especially valid in the assay of substances that are abundantly stored in the platelets, including Sph-1-P. A well-known example in this context is the measurement of the platelet a-granule-derived CXC chemokines, b-TG and platelet factor 4; use of the antiplatelet cocktail CTAD, which potently suppresses platelet intracellular signalling events, 15 and preservation of the samples at 48C were necessary to avoid in vitro platelet activation and accurately assay the plasma concentrations of these CXC chemokines. 22 Therefore, we checked the necessity of careful blood processing for precise measurement of the in vivo plasma Sph-1-P concentration. We determined that preservation and centrifugation of the platelets at 48C were necessary as suitable preparation procedures for the plasma samples to avoid undesired Sph-1-P release in vitro, just as for the CXC chemokine measurement described above.…”

Section: Discussionmentioning

confidence: 99%

“…For example, b-TG, a platelet-specific CXC chemokine stored in and released from the platelet a-granules, is a useful marker for the detection of platelet activation in vivo, to identify patients at risk of thrombosis and for assessing the effects of antiplatelet therapy. In the evaluation of this substance, to avoid an undesired in vitro increase in the plasma b-TG concentration, it is important to use the antiplatelet cocktail CTAD 22 as the anticoagulant, and all the plasma preparation steps should be conducted at 48C. 22 Although EDTA blocks the coagulation cascade and platelet aggregation resulting from the aIIbb3 integrin interaction with fibrinogen, this chelator cannot suppress intracellular signalling events leading to platelet activation, which can be effectively blocked by CTAD.…”

Section: Establishment Of the Sphingosine-1-phosphate Assay Systemmentioning

confidence: 99%

“…In the evaluation of this substance, to avoid an undesired in vitro increase in the plasma b-TG concentration, it is important to use the antiplatelet cocktail CTAD 22 as the anticoagulant, and all the plasma preparation steps should be conducted at 48C. 22 Although EDTA blocks the coagulation cascade and platelet aggregation resulting from the aIIbb3 integrin interaction with fibrinogen, this chelator cannot suppress intracellular signalling events leading to platelet activation, which can be effectively blocked by CTAD. 15 We examined whether these precautions would also be necessary in the preparation of plasma samples for Sph-1-P measurement, since this bioactive lipid is also abundantly stored in platelets and released extracellularly upon activation of the cells.…”

Section: Establishment Of the Sphingosine-1-phosphate Assay Systemmentioning

confidence: 99%

“…EDTA vs. CTAD ( Figure 2a); on the other hand, the concentrations of b-TG were much higher in plasma samples prepared using EDTA than in those prepared using CTAD. 22 We next compared the concentrations of Sph-1-P in plasma samples prepared by centrifugation at 48C and by centrifugation at room temperature; all samples were treated with EDTA þ CTAD (Figure 2b). The plasma concentrations of Sph-1-P were significantly higher when the samples were centrifuged at room temperature: 319.5 + 76.3 nmol/L in samples obtained by centrifugation at 48C vs. 618.0 + 163.1 nmol/L in samples obtained by centrifugation at room temperature.…”

Section: Establishment Of the Sphingosine-1-phosphate Assay Systemmentioning

confidence: 99%

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Plasma sphingosine-1-phosphate measurement in healthy subjects: close correlation with red blood cell parameters

et al. 2008

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Background: Since sphingosine-1-phosphate (Sph-1-P) plays an important role as an extracellular mediator through interaction with specific cell surface receptors, especially in the area of vascular biology and immunology/haematology, determination of its plasma concentration may become important from the clinical viewpoint. Thus, we attempted to develop a method of measuring the plasma Sph-1-P concentration for use in the clinical laboratory setting. Methods: After two-step lipid extraction, Sph-1-P was coupled with o-phthaldialdehyde, and the resultant fluorescent derivative was separated by high-performance liquid chromatography. C 17 -Sph-1-P was used as the internal standard, instead of dihydrosphingosine-1-phosphate, which had been used previously for the same purpose but was actually detected in plasma. Results: Our procedures for preparing the plasma samples and assay Sph-1-P were found to be satisfactory for clinical laboratory testing. The plasma Sph-1-P concentrations were significantly higher in men (413.1 + 52.0 nmol/L; mean + SD) than in women (352.4 + 39.7 nmol/L). Unexpectedly, strong positive correlations were found between the plasma Sph-1-P concentration and red blood cell (RBC)-related parameters, rather than platelet-related parameters. Conclusions: Our present study confirmed the possibility of the clinical introduction of plasma Sph-1-P measurement, and in addition, suggested that RBCs may be involved in the regulation of plasma Sph-1-P concentrations.

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Section: Discussionmentioning

confidence: 99%

Section: Establishment Of the Sphingosine-1-phosphate Assay Systemmentioning

confidence: 99%

Section: Establishment Of the Sphingosine-1-phosphate Assay Systemmentioning

confidence: 99%

Section: Establishment Of the Sphingosine-1-phosphate Assay Systemmentioning

confidence: 99%

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Plasma sphingosine-1-phosphate measurement in healthy subjects: close correlation with red blood cell parameters

et al. 2008

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“…A variety of blood-drawing and samplehandling protocols have been used to isolate PPP. Studies that carefully examined the effect of PPP preparation parameters on observed PPP serotonin concentrations (33,60,62,66,93 ) reported lower concentrations of PPP serotonin [average reported mean for the 5 cited studies was 1.94 (1.09) nmol/L]. Conversely, if it is assumed that lower PPP serotonin values reflect a more accurate estimation of serotonin present in the prepared PPP sample, the effect of sample preparation parameters would be best examined in studies for which lower PPP serotonin concentrations were reported.…”

Section: Measurement Of Platelet-poor Plasma Serotoninmentioning

confidence: 99%

The Measurement of Platelet-Poor Plasma Serotonin: A Systematic Review of Prior Reports and Recommendations for Improved Analysis

2011

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BACKGROUND: Recent reports of new and important roles for serotonin (5-hydroxytryptamine, 5-HT) in the periphery have substantially increased interest in measuring peripheral serotonin. Nearly all circulating serotonin is found within platelets and this pool has been assessed by measuring serotonin in whole blood or in platelet-rich plasma. Measurement of the much smaller but potentially critically important pool of human free plasma serotonin in plateletpoor plasma (PPP) has proven much more difficult, with a wide range of reference values reported. CONTENT:To characterize the available data we carried out a systematic literature search of previous reports of PPP serotonin and attempted to determine the best estimate of true PPP serotonin concentration in humans. A total of 101 published reports that included PPP serotonin values in healthy controls were found and included in the summary statistical analyses. The distribution of PPP serotonin values demonstrated high skewness (ϩ1.98), and the reported values ranged from 0.6 to 179 nmol/L, with a mean of 31.6 nmol/L, an SD of 38.9 nmol/L, and a median of 14.8 nmol/L.

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Determination of free serotonin and its metabolite 5‐HIAA in blood human samples with consideration to pre‐analytical factors

Yubero-Lahoz

Rodríguez

Faura

et al. 2014

Biomedical Chromatography

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Significant differences have been reported over the years in measuring physiological levels of free circulating serotonin (f5-HT) in platelet-poor plasma (PPP). This work shows that there are crucial pre-analytical factors in sample manipulation that can provoke an artifactual release of 5-HT from platelets, and that, even when the sample is accurately processed to obtain PPP, f5-HT levels are approximately 2.8 times higher than those of f5-HT in blood. An alternative methodology consisting of ex vivo blood microdialysis coupled to high-performance liquid chromatography-electrochemical detection is proposed and validated. It is considered the most accurate technique to measure physiological circulating f5-HT and its metabolite 5-hydroxyindoleacetic acid (f5-HIAA), owing to its sensitivity (limits of quantification of 0.08 ng/mL) and reliability since there is no sample manipulation. The f5-HT and f5-HIAA levels in blood and in PPP were studied in control subjects, hypertensive and end-stage renal disease patients, who have a deregulated serotonergic system. This work reveals that blood is the best matrix to determine f5-HT concentrations, and the clinical relevance of the accuracy of f5-HT determination is discussed.

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Platelet release of β-thromboglobulin and platelet factor 4 and serotonin in plasma samples

Cited by 37 publications

References 10 publications

Plasma sphingosine-1-phosphate measurement in healthy subjects: close correlation with red blood cell parameters

Plasma sphingosine-1-phosphate measurement in healthy subjects: close correlation with red blood cell parameters

The Measurement of Platelet-Poor Plasma Serotonin: A Systematic Review of Prior Reports and Recommendations for Improved Analysis

Determination of free serotonin and its metabolite 5‐HIAA in blood human samples with consideration to pre‐analytical factors

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