Transfer of arachidonic acid from phosphatidylcholine to phosphatidylethanolamine during storage of human platelets for 5 days

1990 ◽  
Vol 68 (1) ◽  
pp. 117-122 ◽  
Author(s):  
Julie Lacasse ◽  
Rosalind S. Labow ◽  
Morris Kates ◽  
George A. Adams
Keyword(s):  
Arachidonic Acid ◽  
Storage Conditions ◽  
Phospholipid Metabolism ◽  
Human Platelets ◽  
Acyl Transfer ◽  
Acid Uptake ◽  
Molar Composition ◽  
Platelet Storage ◽  
Head Groups ◽  
Glycerol Uptake

Human platelets are routinely stored for 5 days prior to transfusion, but they deteriorate during storage. Since very little information is available concerning the effect of storage on platelet phospholipid metabolism, the biosynthesis and remodelling of platelet phospholipids were studied. Platelets were incubated separately with [14C]glycerol, [14C]arachidonic acid, or a mixture of [14C]glycerol and [3H]arachidonic acid, and stored in a platelet storage medium at 22 °C. Maximum glycerol uptake (20%) was attained after 6 h. [14C]Glycerol was incorporated into phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol, and to a much lesser extent phosphatidylserine, under storage conditions for 5 days. The distribution of the initial arachidonic acid uptake was not as would be expected based on the molar composition of endogenous phospholipids. The arachidonic acid (75%) which was taken up within 10 min of incubation distributed 55% into the phosphatidylcholine and only 14% into the phosphatidylethanolamine; the molar composition is actually 18% phosphatidylcholine and 47% phosphatidylethanolamine. During storage, there was a continuous transfer of the radiolabeled arachidonic acid from phosphatidylcholine to phosphatidylethanolamine until, after 5 days, the distribution of arachidonic acid was identical to the endogenous distribution. In contrast, no change in the glycerol incorporation pattern was detected during storage. This suggested that the mechanism for arachidonic acid redistribution was not through exchange of polar head groups, but through acyl transfer of arachidonic acid from phosphatidylcholine to phosphatidylethanolamine.Key words: human, platelet, storage, arachidonate, phospholipids.

Arachidonic acid uptake by human platelets is mediated by CD36

Platelets ◽  
1996 ◽  
Vol 7 (5-6) ◽  
pp. 291-295 ◽  
Author(s):  
A. K. Dutta-Roy ◽  
M. J. Gordon ◽  
F. M. Campbell ◽  
L. C. Crosbie
Keyword(s):  
Arachidonic Acid ◽  
Human Platelets ◽  
Acid Uptake

Altered phospholipid metabolism in thrombin-stimulated human platelets

1984 ◽  
Vol 62 (6) ◽  
pp. 341-351 ◽  
Author(s):  
Bruce J. Holub
Keyword(s):  
Arachidonic Acid ◽  
Species Composition ◽  
Phospholipid Metabolism ◽  
Human Platelets ◽  
Inositol Triphosphate ◽  
Molecular Species Composition ◽  
The Past ◽  
Free Arachidonic Acid ◽  
The Individual ◽  
Individual Enzyme

Extensive research during the past few years has indicated that dramatic alterations in phospholipid metabolism and composition represent early and important biochemical events in the response of human platelets to thrombin stimulation. The individual enzyme-catalyzed steps which provide for the release of free arachidonic acid for thromboxane A2 formation via the initial degradation of phosphatidylcholine and phosphatidylinositol have been studied. Their importance in this regard is influenced by the molecular species composition of the corresponding phospholipid precursors. A role for stimulated phosphatidylinositol 4,5-bisphosphate degradation in the phosphatidylinositol response and inositol triphosphate release associated with calcium mobilization has also been proposed. The 1,2-diacylglycerol released by the action of phospholipase C on phosphatidylinositol and its 4,5-bisphosphate derivative has been implicated as an activator of protein phosphorylation; the derived phosphatidic acid has been proposed as a mediator for promoting an intracellular flux of calcium associated with platelet responses.

Effect of Dilazep on Phospholipid Metabolism and Arachidonic Acid Cascade in Human Platelets in vitro

1984 ◽  
Vol 12 (2) ◽  
pp. 363-370
Author(s):  
Mitsuko TAKENAGA ◽  
Haruo KITAGAWA ◽  
Aizan HIRAI ◽  
Yasushl TAMURA ◽  
Sho YOSHIDA
Keyword(s):  
Arachidonic Acid ◽  
Phospholipid Metabolism ◽  
Human Platelets ◽  
Arachidonic Acid Cascade

Loss of Thrombin-Induced Ca2+ Mobilization in a Subpopulation of Platelets during Storage

1991 ◽  
Vol 66 (03) ◽  
pp. 350-354 ◽  
Author(s):  
Rob Fijnheer ◽  
Christa H E Homburg ◽  
Berend Hooibrink ◽  
Martine N Boomgaard ◽  
Dirk de Korte ◽  
...  
Keyword(s):  
Human Platelets ◽  
Flow Cytometer ◽  
Maximal Concentration ◽  
Gradual Decline ◽  
Platelet Storage ◽  
Functional Defect ◽  
Total Fluorescence ◽  
Induced Changes

SummaryThrombin-induced changes in cytosolic free Ca2+ ([Ca2+]i) were studied in human platelets that had been stored for up to 6 days. Changes in [Ca2+]i were measured with Indo-1-loaded platelets and quantitated with two different methods: (i) measurement of the changes in total fluorescence; (ii) measurement of the [Ca2+]i changes in individual platelets in a flow cytometer, allowing the detection of non-responding platelets. The maximal concentration of [Ca2+]i after stimulation with 0.5 U of thrombin/ml decreased from 544 ± 58 nM (mean ± SEM, n = 6) on day 0, to 276 ± 9 nM on day 3 and to 203 ± 23 nM on day 6. The percentage of platelets responding to 0.5 U of thrombin/ml declined from 90 ± 2% on day 0 to 72 ± 4% on day 3, and to 47 ± 8% on day 6. Nevertheless, also the responding platelets showed a decreased rise in [Ca2+]i.The study shows that during platelet storage a decrease in the rise in [Ca2+]i upon thrombin stimulation occurs. This decrease is partly due to the formation of a subpopulation of platelets that is completely unresponsive and partly due to a decreased responsiveness in the remainder of the platelets; it is not due to a gradual decline in [Ca2+]i rise in all platelets. This phenomenon provides new insight in the functional defect of stored platelets.

Antithrombin III Antigen in Human Platelets

1985 ◽  
Vol 54 (03) ◽  
pp. 599-602 ◽  
Author(s):  
M Léon Alhenc-Gelas ◽  
M Aiach ◽  
A Gorenflot ◽  
J P Andreux
Keyword(s):  
Arachidonic Acid ◽  
Enzyme Immunoassay ◽  
Antithrombin Iii ◽  
Human Platelets ◽  
Mean Value ◽  
Normal Blood ◽  
Freezing And Thawing ◽  
Healthy Donors ◽  
At Iii ◽  
Storage Granules

SummaryImmunoreactive AT III was found in human platelets. AT III antigen was quantified in platelets taken from each of 17 healthy donors by a specific competitive enzyme immunoassay using purified AT III and AT III antibodies. AT III antigen levels in extracts of washed platelets disrupted by freezing and thawing ranged from 32 to 140 ng per 109 platelets with a mean value of 70.3 ± 27.3. When stimulated by arachidonic acid, the platelets released AT III antigen together with immunoreactive fibrinogen. These results show that AT III is present in platelets at a level corresponding to approximately 0.01% of total antithrombin in normal blood, and suggest that platelet AT III, like fibrinogen, is contained in the storage granules.

Pretreatment of Human Platelets with Plasmin Inhibits Responses toThrombin, but Potentiates Responses to Low Concentrations of Aggregating Agents, Including the Thrombin Receptor Activating Peptide, SFLLRN

1997 ◽  
Vol 77 (04) ◽  
pp. 741-747 ◽  
Author(s):  
R L Kinlough-Rathbone ◽  
D W Perry ◽  
M L Rand ◽  
M A Packham
Keyword(s):  
Arachidonic Acid ◽  
Platelet Activating Factor ◽  
Human Platelets ◽  
Thrombin Receptor ◽  
Fibrinolytic Agents ◽  
Albumin Solution ◽  
Fibrinogen Receptor ◽  
Low Concentrations ◽  
Induced Aggregation

SummaryEffects of plasmin on platelets, that influence subsequent responses to aggregating agents, are relevant to attempts to prevent rethrombosis following administration of fibrinolytic agents. We describe plasmin-induced inhibition of platelet responses to thrombin, but potentiation of responses to other aggregating agents. Washed human platelets were labeled with 14C-serotonin, treated for 30 min at 37° C with 0, 0.1 or 0.2 CU/ml of plasmin, followed by aprotinin, washed and resuspended in a Tyrode-albumin solution with apyrase. Incubation with 0.2 CU/ml of plasmin almost completely inhibited thrombin-induced (0.1 U/ml) aggregation, release of 14C-serotonin, and increase in cytosolic [Ca2+]. In contrast, with plasmin-pretreated platelets, aggregation and release of 14C-serotonin were strongly potentiated in response to low concentrations of the thrombin receptor-activating peptide SFLLRN, ADP, platelet-activating factor, collagen, arachidonic acid, the thromboxane mimetic U46619, and the calcium ionophores A23187 and ionomycin. Aspirin or RGDS partially inhibited potentiation. Plasmin-pretreated platelets resuspended in plasma anticoagulated with FPRCH2C1 (PPACK) also showed enhanced responses to aggregating agents other than thrombin. The contrasting effects on responses to thrombin and SFLLRN are noteworthy. Plasmin cleaves GPIIb/IIIa so that it becomes a competent fibrinogen receptor, and binding of 125I-fibrinogen during ADP-induced aggregation was greatly potentiated within 10 s. Potentiation of aggregation by other agonists may be due to increased binding of released fibrinogen. Thus, platelets freed from a thrombus may have increased responsiveness to low concentrations of aggregating agents other than thrombin. These results provide further support for the use of inhibitors of platelet reactions in conjunction with administration of fibrinolytic agents.

Correlation Between Noradrenaline Fluxes, Metabolism and Compartmentalisation in Human Platelets

1982 ◽  
Vol 48 (01) ◽  
pp. 062-066 ◽  
Author(s):  
Chantal Legrand ◽  
Véronique Dubernard ◽  
Philippe Meyer
Keyword(s):  
Noradrenaline Release ◽  
Human Platelets ◽  
Release Of Noradrenaline ◽  
Storage Pool ◽  
Platelet Storage ◽  
Efflux Of Noradrenaline ◽  
Preferential Localization

Summary(3H) noradrenaline was taken up by human platelets and partially converted into sulfoconjugated noradrenaline. This uptake was inhibited by drugs which have been previously shown to impair the uptake of 5-HT (ouabain, chlorimipramine) or the storage of 5-HT (tyramine, reserpine) by platelets. In addition, tyramine and reserpine stimulated the formation of sulfoconjugated noradrenaline. The efflux of noradrenaline from platelets was measured in parallel and was found to be directly related to the proportion of non metabolized to metabolized noradrenaline in the cells. Unlike tyramine, which induced a similar release of noradrenaline and 5-HT, reserpine was less effective at inducing noradrenaline release than 5-HT release. This study indicates a preferential localization of noradrenaline in the granular pool of human platelets with the existence of an extragranular sulfoconjugated pool which is increased when the granular storage of noradrenaline is impaired. Studies of noradrenaline fluxes and metabolism may be useful in the understanding of both acquired and inherited platelet storage pool defects.

Effects of Dipyrone on Prostaglandin Production by Human Platelets and Cultured Bovine Aortic Endothelial Cells

1983 ◽  
Vol 49 (02) ◽  
pp. 132-137 ◽  
Author(s):  
A Eldor ◽  
G Polliack ◽  
I Vlodavsky ◽  
M Levy
Keyword(s):  
Endothelial Cells ◽  
Arachidonic Acid ◽  
Competitive Inhibition ◽  
Inhibitory Effect ◽  
Human Platelets ◽  
Ionophore A23187 ◽  
Aortic Endothelial Cells ◽  
Bovine Aortic Endothelial Cells ◽  
Aortic Endothelial

SummaryDipyrone and its metabolites 4-methylaminoantipyrine, 4-aminoantipyrine, 4-acetylaminoantipyrine and 4-formylaminoan- tipyrine inhibited the formation of thromboxane A2 (TXA2) during in vitro platelet aggregation induced by ADP, epinephrine, collagen, ionophore A23187 and arachidonic acid. Inhibition occurred after a short incubation (30–40 sec) and depended on the concentration of the drug or its metabolites and the aggregating agents. The minimal inhibitory concentration of dipyrone needed to completely block aggregation varied between individual donors, and related directly to the inherent capacity of their platelets to synthesize TXA2.Incubation of dipyrone with cultured bovine aortic endothelial cells resulted in a time and dose dependent inhibition of the release of prostacyclin (PGI2) into the culture medium. However, inhibition was abolished when the drug was removed from the culture, or when the cells were stimulated to produce PGI2 with either arachidonic acid or ionophore A23187.These results indicate that dipyrone exerts its inhibitory effect on prostaglandins synthesis by platelets or endothelial cells through a competitive inhibition of the cyclooxygenase system.

In Vitro Incorporation and Metabolism of Icosapentaenoic and Docosahexaenoic Acids in Human Platelets - Effect on Aggregation

1986 ◽  
Vol 56 (01) ◽  
pp. 057-062 ◽  
Author(s):  
Martine Croset ◽  
M Lagarde
Keyword(s):  
Arachidonic Acid ◽  
Fatty Acid ◽  
Platelet Aggregation ◽  
Fatty Acid Distribution ◽  
Thromboxane A2 ◽  
Human Platelets ◽  
Aggregation Response ◽  
Docosahexaenoic Acids ◽  
Membrane Level

SummaryWashed human platelets were pre-loaded with icosapentaenoic acid (EPA), docosahexaenoic acid (DHA) or EPA + DHA and tested for their aggregation response in comparison with control platelets. In fatty acid-rich platelets, an inhibition of the aggregation could be observed when induced by thrombin, collagen or U-46619. The strongest inhibition was observed with DHA-rich platelets and it was reduced when DHA was incorporated in the presence of EPA.Study of fatty acid distribution in cell lipids after loading showed that around 90% of EPA or DHA taken up was acylated into phospholipids and a very small amount (less than 2%) remained in their free and hydroxylated forms. DHA was more efficiently acylated into phosphatidylethanolamine (PE) than into phosphatidylinositol (PI) in contrast to what observed with EPA, and both acids were preferentially incorporated into phosphatidylcholine (PC). EPA inhibited total incorporation of DHA and increased its relative acylation into PE at the expense of PC. In contrast, DHA did not affect the acylation of EPA. Upon stimulation with, thrombin, EPA was liberated from phospholipids and oxygenated (as judged by the formation of its monohydroxy derivative) whereas DHA was much less metabolized, although consistently transferred into PE.It is concluded that EPA and DHA might affect platelet aggregation via different mechanisms when pre-loaded in phospholipids. Whereas EPA is known to alter thromboxane A2 metabolism from endogenous arachidonic acid, by competing with it, DHA might act directly at the membrane level for inhibiting aggregation.

scholarly journals Platelet storage and functional integrity

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Gianmatteo Vit ◽  
Harald Klüter ◽  
Patrick Wuchter
Keyword(s):  
Platelet Transfusion ◽  
Structural Characteristics ◽  
Storage Conditions ◽  
Transfusion Practice ◽  
Common Interest ◽  
Laboratory Staff ◽  
German Medical Association ◽  
Platelet Storage ◽  
Technological Advances ◽  
Assessment Procedures

AbstractPlatelet transfusion is a topic of common interest for many specialists involved in patient care, from laboratory staff to clinical physicians. Various aspects make this type of transfusion different from those of other blood components. In this review, the challenges in platelet transfusion practice that are relevant for laboratory colleagues will be discussed, highlighting how the biochemical and structural characteristics of these blood elements directly affect their function and consequently the clinical outcome. More than 1,300 platelet concentrates are transfused in Germany every day, and several types are offered by their respective manufacturers. We describe the technological advances in platelet concentrate production, with a focus on how the storage conditions of platelets can be improved. Laboratory quality assessment procedures for a safe transfusion are discussed in detail. For this purpose, we will refer to the Hemotherapy Directives (Richtlinie Hämotherapie) of the German Medical Association.

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