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Platelets expressing P-selectin and platelet-derived microparticles in stored platelet concentrates bind to PSGL-1 on filtrated leukocytes.

The levels of interleukin-6 and platelet-derived microparticles (PMPs) were measured in the blood of 137 patients with side effects from platelet concentrate (PC) transfusion with leukocyte removal filtration, P-selectin-expressing platelet and PMPs in stored PC before and after the filtration, and filtered leukocytes positive for P-selectin glycoprotein ligand-1. The side effects, which were observed in 203 transfusions for 84 patients with hematologic disease and 53 patients with nonhematologic disease with no significant difference between the two groups, included urticaria (75.9%), erythema (18.7%), and fever (17.2%), but no anaphylactic reactions. The levels of interleukin-6 and PMP correlated in both groups, and were significantly higher in the hematologic disease group than in the nonhematologic disease group. The level of PMP, but not interleukin-6, was significantly higher for patients testing positive for allergic reaction than for those testing negative. In the stored PC prior to filtration, the level of interleukin-6 was normal. The level of P-selectin-expressing platelets and PMPs was elevated before filtration, but was significantly lower after filtration. Taken together, the results suggest that PMP is involved in the generation of transfusion reactions, and indicate that both platelets and PMP displaying P-selectin bind to P-selectin glycoprotein ligand-1 of leukocytes retained by the leukocyte filter.

Blood Platelets↗

Platelet deposition at angioplasty sites and platelet survival time after PTA in iliac and femoral arteries: investigations with indium-111-oxine labelled platelets in patients with ASA (1.0 g/day)-therapy.

In this study we have assessed the deposition of 111-In-oxine-labelled platelets--using a dual radiotracer method--at angioplasty sites of the lower extremities in 20 patients (14 male, 6 female; median age: 60 years) with ASA (1.0 g/day)-therapy. The platelet survival time (PST)--using the multiple hit model--was evaluated before and after percutaneous transluminal angioplasty, and we also measured the plasma levels of beta-thromboglobulin (beta-TG) and platelet factor 4 (PF 4) before and after PTA. Before PTA, scintigraphy was positive in only one patient, while 24 hours after PTA a positive scintigraphic result was observed in 16/20 patients. The median target/non target-ratio was 1.0 (0.66-1.3) before PTA, and this ratio increased significantly (p less than 0.0005) to 1.53 (1.0-3.3) after PTA. The median PST decreased significantly (185.0 hours before PTA----145.2 hours after PTA; p less than 0.001), while the median platelet turnover increased from 34,000/microliter/day to 47,900/microliter/day (p less than 0.01). The median plasma levels of the platelet specific proteins increased significantly immediately after PTA (p less than 0.001), but one day later they were not significantly different from the pretreatment values. The quantitative methods used in this study seem a valuable tool to evaluate the effects of different therapeutical--especially antiplatelet--interventions after PTA in humans, thus helping to find the best antithrombotic regimen for this widely used therapeutical procedure.

Adult↗

Combined platelet-leukapheresis: a technique for preparing separate platelet and granulocyte-platelet units from single donors.

One donor can provide separate platelet and granulocyte-platelet units that have the potential to fulfill the needs of two recipients. Combined platelet and leukapheresis were performed in the presence of hyddroxyethyl starch and citrate employing the Haemonetics Model 30 Blood Separator. Platelets and granulocytes were separated subsequently by centrifugation and two individual units were prepared for transfusion. The platelet units contained a mean of 4.4 x 10(11) platelets/unit and were nearly devoid of leukocytes. The combined granulocyte-platelet units contained a mean of 7.38 x 10(9) neutrophils/unit and 3.06 x 10(11) platelets/unit. Thus two separate units containing sufficient numbers of cells for transfusion can be obtained from pheresis of one donor.

Blood Donors↗

Automated analysis of feline platelets in whole blood, including platelet count, mean platelet volume, and activation state.

A new whole-blood flow cytometric method has been developed for counting and sizing platelets in samples from cats, a species in which platelet and red blood cell sizes overlap significantly. The method is a modified version of the two-angle laser light scattering technology used by Bayer H*System hematology analyzers. The new method provided accurate platelet counts and mean platelet volumes (MPV, fl) for cats. The method also measured mean platelet component concentration (MPC, g/dl), a parameter which was shown to be sensitive to platelet activation state, and which decreased in value as activation progressed.

Journal Article↗

Platelet storage lesion: formation of platelet fragments with platelet factor 3 activity.

Certain samples of stored platelet concentrates exhibited marked platelet factor 3 activity. This activity was associated almost completely with platelet fragments formed during storage. Platelet concentrates containing high levels of these fragments and hence high platelet factor 3 activity were characterized after storage by alkaline plasma pH values, high levels of extracellular lactate dehydrogenase activity and relatively low cell counts.

Blood Platelets↗

[Anti-leuko-platelet allo-immunization in blood replacements. Do transfusions of platelets from single donors present an advantage over platelets from multiple donors?].

42 patients with acute leukaemia, treated with cytotoxic drugs, have been evaluated retrospectively: --group I: 11 patients received packed red blood cells and platelets from single donors; --group II: 6 patients received packed red blood cells and platelets from multiple donors; --group III: 25 patients received packed red blood cells and platelets from single or multiple donors and granulocytes transfusions. There was no difference in age, sex, time of follow up, number of transfusions, in the three groups. The rate of alloimmunization defined as lymphocytotoxicity against more than 20% of a panel of 24 lymphocytes, was 33% (36% group I--33% group II--32% group III). This study shows that platelets from single donors are of no use in preventing or delaying alloimmunization. On the other hand, their major interest is to provide alloimmunized patients with compatible platelets.

Adolescent↗

Activation of platelets by platelet activating factor (PAF) derived from IgE-sensitized basophils. IV. PAF does not activate platelet factor 3 (PF3).

Platelet activating factor (PAF) derived from antigen-stimulated, IgE-sensitized rabbit basophils acts on platelets to induce aggregation and secretion of their content of granule-bound vasoactive amines. Despite this, PAF did not activate platelet factor 3. In contrast, collagen induced aggregation, secretion and PF3 activation in the washed platelets. Other stimuli (ADP, C3b, thrombin) also initiated both secretion and PF3 activation. A wide dose range of PAC, including those giving maximal secretion and aggregation, were ineffictive in making PF3 available and the possibility that PAF inhibited PF3, or its generation, was also excluded. It is concluded that PAF is a unique stimulus for platelets and that secretion and aggregation are not necessarily accompanied by PF3 generation.

Animals↗

Smooth muscle cell migration induced by shear-loaded platelets and endothelial cells. Enhanced platelet-derived growth factor production by shear-loaded platelets.

BACKGROUND: Uncontrolled migration and proliferation of smooth muscle cells (SMC) are the main mechanisms of development of atherosclerotic neointima. However, it has not yet been elucidated how mechanical stress is involved in this process. We investigated smooth muscle cell mitogenic activity induced by shear-loaded platelets and endothelial cells. METHODS EXPERIMENTAL DESIGN: in vitro experimental study. We devised four types of conditioned media; supernatant of mixed culture of platelets and endothelial cell (ST), supernatant of shear-loaded mixed culture of platelets and endothelial cell (SH), ST medium neutralised with anti-PDGF antibody (ST+), and SH medium neutralised with anti-PDGF antibody (SH+). Smooth muscle cells were cultured in each conditioned medium, and their spreading activity was determined under a microscope. RESULTS: Smooth muscle cells spreading activity in the SH group was significantly greater than that in the ST group. Their spreading activity was suppressed by anti-PDGF antibody under shear conditions (SH+), but it was not by anti-PDGF antibody under static conditions (ST+). CONCLUSIONS: Our results demonstrate that platelet-derived growth factor is produced by shear-loaded platelets and endothelial cells, and local mechanical forces may play an important role in cardiovascular disease.

Blood Platelets↗

[Effect of dietary linoleic acid on the platelet aggregation, fatty acid composition of platelet phospholipids and synthesis of platelet prostaglandins in rats].

Three groups of Wistar-Kyoto rats were kept on diets containing different quantity of linoleic acid (LA) in prenatal and postnatal periods up to 18 weeks age: I. less than 0.1 cal.% of LA, 2. 9.0 cal.% of LA and 3. 16 cal.% of LA. Aggregation of washed platelets with ADP and A23187, biosynthesis of prostaglandins from 14C-arachidonic acid in platelets and fatty acid composition of platelet phospholipids were studied using gas liquid chromatography. Modification of fatty acid composition of platelet phospholipids and changes in biosynthesis of platelet prostaglandins are the base for proaggregation effect of LA deficient and for antiaggregation effect of LA enriched diets. The effects of dietary LA were mediated not only via altered metabolism of fatty acids of linoleic family but also via metabolism of fatty acids of oleic and linolenic series.

Animals↗

Differential in vitro effects of the platelet glycoprotein IIb/IIIa inhibitors abixicimab or SR121566A on platelet aggregation, fibrinogen binding and platelet secretory parameters.

The aim of this study was to compare fibrinogen binding, inhibition of platelet aggregation and secretory potential of the MAb abciximab (0.5-5 microg/mL) and the peptidomimetic compound SR121566A (15-250 ng/mL) in vitro in whole blood. Fibrinogen binding was followed by flow cytometry; platelet function was evaluated by light transmittance and by impedance aggregometry. Secretory functions of platelets were evaluated using ATP as marker for early secretion by dense granulae and P-selectin (CD62) for alpha-granular secretion as well as CD63 for lysosomal degranulation. Results showed that fibrinogen binding induced by 5 microM TRAP was maximally inhibited greater than 80% at 3 microg/mL abciximab or at 250 ng/mL SR121566A. At these concentrations of antagonists, platelet aggregation induced by 5 microM ADP or 2 microg/mL collagen was inhibited completely. Expression of CD62 was reduced 34% with abciximab or 15% with SR121566A; CD63 expression was reduced 22% with both agents. With both agents, the EC50 for inhibition of CD62 and CD63 expressions was in similar magnitudes than the EC50 for fibrinogen binding inhibition. With 3 microg/mL abciximab, ATP secretion was maximally reduced to 50% of the control, whereas SR121566A at 250 ng/mL had no inhibitory effect on this parameter. A slight increase in ATP secretion was seen with 0.5 microg/mL abciximab and with SR121566A in concentrations of less than 45 ng/mL. The data suggest a discoupling between the anti-aggregatory and the antisecretory effects of IIb/IIIa antagonists. Because it is not established to what extend CD62 or CD63 expression can be reduced by any means, the reduction by 20-30% obtained by 3 microg/mL abciximab or 250 ng/mL SR121566A might already be the maximum possible inhibition by these agents.

Adenosine Triphosphate↗

Effect of platelet agonists on the binding of platelet-activating factor to human platelets.

The binding of PAF in vitro to its receptors on platelets gradually becomes irreversible. This property has been applied to the measurement of PAF in circulating blood, the concept being that in vivo contact with endogenous PAF diminishes subsequent in vitro binding of 3H-labeled PAF. Here we show that, when platelets are stimulated with thrombin, the subsequent binding of [3H]PAF is greatly reduced, probably due to proteolytic damage of the PAF receptors. In addition, [3H]PAF binding is decreased, when the cell's energy status is low, which is seen during thrombin-induced secretion. Thus, the detection of PAF in circulating blood based on the binding to PAF receptors on platelets is severely hampered when platelets make contact with thrombin.

Adenosine Diphosphate↗

Combined effects of mild hypothermia and glycoprotein IIb/IIIa antagonists on platelet-platelet and leukocyte-platelet aggregation.

Endovascular cooling was assessed as a potential treatment for percutaneous coronary intervention in patients with acute myocardial infarction. Here we show that mild hypothermia: (1) inhibits platelet aggregation; (2) augments eptifibatide- and tirofiban- but not abciximab-induced inhibition of platelet aggregation; (3) increases the formation of adenosine diphosphate-induced leukocyte-platelet aggregates; and (4) diminishes the glycoprotein IIb/IIIa antagonist-induced decrease in leukocyte-platelet aggregates.

Abciximab↗

Effects of a thio analog of platelet-activating factor on platelet aggregation and adenosine 3',5'-monophosphate concentration in hepatocyte suspensions and in platelets. A comparison with the naturally occurring compound.

2-Acetyl-S-octadecyl-rac-1-thioglycero-3-phosphocholine, the thio analog of platelet-activating factor, in concentrations of 10(-6) M to 10(-12) M in the medium, lowers cAMP-concentrations in rat hepatocytes. In a concentration of 10(-4) M, the thio analog aggregates human platelets irreversibly, above 10(-5) M aggregation shows a reversible course. Compared with the saturated or unsaturated ether analogs, the thio compound shows less platelet-aggregating potency. We have correlated this difference in platelet aggregation with reduced cAMP-depressing activity of the thio analog (compared with saturated and unsaturated 2-acetyl-1-O-alkyl-sn-glycero-3-phosphocholines).

Alprostadil↗

Selective modulation of the human platelet thromboxane A2/prostaglandin H2 receptor by eicosapentaenoic and docosahexaenoic acids in intact platelets and solubilized platelet membranes.

We previously demonstrated that nonesterified as well as esterified eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3) inhibit U46619-induced platelet aggregation and [3H]U46619 specific binding to washed human platelets. It was also demonstrated that esterification of these fatty acids resulted in a decrease in the affinity of [3H]U46619 for the thromboxane A2/prostaglandin H2 (TXA2/PGH2) receptor. In order to investigate the specificity of this inhibition, the effects of 20:5n-3 and 22:6n-3 on the function and binding of the platelet alpha 2-adrenergic receptor were studied. It was found that neither 20:5n-3 nor 22:6n-3 (nonesterified or esterified) altered epinephrine-induced aggregation or [3H]yohimbine specific binding. Moreover, Scatchard analysis revealed that esterification with either 20:5n-3 or 22:6n-3 did not alter the dissociation constant for [3H]yohimbine binding. Modulation of the TXA2/PGH2 receptor by 20:5n-3 and 22:6n-3 was next evaluated using CHAPS- and digitonin-solubilized platelet membranes. [3H]SQ29,548 dissociation constants of 26.5 nM and 20.8 nM were measured for CHAPS and digitonin-solubilized membranes, respectively. Competitive binding experiments in these solubilized preparations revealed that 20:5n-3 or 22:6n-3 blocked [3H] SQ29,548 binding with IC50 values in the range of 6-15 microM, while concentrations of these fatty acids of up to 100 microM showed no effect on [3H]yohimbine binding. On the other hand, the IC50 values for inhibition of [3H] SQ29,548 binding by linoleic acid (18:2n-6) and gamma-linolenic acid (18:3n-6) were in the range of 150 microM. Furthermore, 18:2n-6 and 18:3n-6 showed similar inhibitory effects on [3H]yohimbine binding. Finally, competition binding studies performed in a partially purified TXA2/PGH2 receptor preparation also demonstrated inhibition of [3H]SQ29,548 binding by 20:5n-3 and 22:6n-3. Collectively, these findings support the notion that 20:5n-3 and 22:6n-3 can selectively and directly modulate TXA2/PGH2 receptor function, and that this mechanism of action may contribute to the antiplatelet activity associated with diets rich in these fatty acids.

Blood Platelets↗

Comparison of the direct platelet immunofluorescence test (direct PIFT) with a modified direct monoclonal antibody-specific immobilization of platelet antigens (direct MAIPA) in detection of platelet-associated IgG.

Glycoprotein (GP)-specific platelet-associated IgG (PA-IgG) may be demonstrable in autoimmune-mediated thrombocytopenia. We studied 159 consecutive patients with histories of thrombocytopenia by a modified direct monoclonal antibody-specific immobilization of platelet antigens (direct MAIPA) assay, which immobilizes GP IIb/ IIIa, GP Ib/IX and GP Ia/IIa simultaneously. This modification requires smaller quantities of platelets than standard measurements performed separately. PA-IgG was present in 84/159 (53%) patients, as shown by the direct platelet immunofluorescence test (PIFT) with flow cytometry as a reference. PA-IgG against GP IIb/IIIa and/or GP Ib/IX and/or GP Ia/IIa was noted in 46 patients (29%), of whom 93% (43/46)-were also PA-IgG positive. The amount of PA-IgG detected by PIFT correlated directly with that detected by direct MAIPA (r = 0.71; P < 0.001). Only three patients 12548 with negative direct PIFT had GP-specific PA-IgG. GPV-specific PA-IgG was detected in 13 (10%) of the 125 patients, in whom further studies could be performed. In the subgroup of patients with GP-specific PA-IgG, the median fluorescence intensities of direct PIFT were higher than in patients with no GP-specific PA-IgG (P < 0.001). Direct PIFT and direct MAIPA divided the patients into asymmetric subgroups. However, the relative roles of these tests in the diagnosis of autoimmune-mediated thrombocytopenia await further studies.

Adolescent↗

Platelet associated IgG, platelet survival, and platelet sequestration in thrombocytopenic states.

Platelet-associated IgG (PAIgG), platelet mean life span (MLS), and platelet sequestration sites were studied in 69 patients with immune (ITP) and presumed nonimmune thrombocytopenias (NTP). A shortened MLS was associated with elevated PAIgG (N = 46), and with normal PAIgG (N = 15). Four patients had a normal MLS, but elevated PAIgG, four patients were normal for both parameters. The highest PAIgG values occurred in ITP patients with a very short MLS. Nine NTP patients had also elevated PAIgG, but a normal or slightly shortened MLS. There was a significant double log correlation between PAIgG and MLS for ITP, but not for NTP patients. Judged from the coefficient of determination, only 10% of PAIgG were directly related to a shortened MLS. 70% of patients (N = 63) had exclusively splenic and 30% hepatosplenic sequestration. PAIgG was elevated in 29/44 patients with splenic (66%) and in 16/19 patients with hepatosplenic sequestration (84%). In ITP, PAIgG-positive cases were observed in 69% of splenic v 82% of hepatosplenic sequestration, while in NTP the corresponding figures were 6/11 and 2/2. No significant correlation between PAIgG and either sequestration type was demonstrable. We conclude that in immunologically mediated thrombocytopenia only a small portion of PAIgG accounts for a decreased MLS, and that the concentration of PAIgG per se does not determine the platelet sequestration type.

Adolescent↗

The platelet count accuracy of platelet concentrates obtained by using automated analysis is influenced by instrument bias and activated platelet components.

BACKGROUND AND OBJECTIVES: The blood platelet content (in numbers) of platelet concentrates is required for production quality control and to predict clinical responses. MATERIALS AND METHODS: This study compared the performance of automated counting from impedance and optical instruments to data from immunoplatelet reference analysis. RESULTS: All methods showed good linearity with evidence of significant instrument-specific deviations from the line of agreement. Relational formulae largely corrected bias, but did not resolve platelet count variability. A second confounding factor, related to the proportion of small (activated) platelets, was also shown to contribute to intermethod discrepancies. CONCLUSIONS: Blood processing centres should establish correction factors for each instrument compared to reference methods, such as the immunoplatelet count.

Analysis of Variance↗

Modulation of platelet function by prostaglandins: characterization of platelet receptors for stimulatory prostaglandins and the role of arachidonate metabolites in platelet degranulation responses.

The effects of PGG2 and PGH2 on platelets are mimicked by synthetic PG analogues in which the nature and specificity of the substituents on carbons 11 and 15 (or 16) are important determinants of reactivity. Arachidonic acid and stimulatory PGs induce secretion of platelet dense granule and alpha granule constituents, but not lysomal constituents, although arachidonate metabolism is necessary for collagen-induced release of lysosomal enzymes. NO164 acts on platelets as an endoperoxide antagonist: Trimethoquinol acts as an endoperoxide and TxA2 antagonist. PGs induce platelet aggregation by combining with a specific (endoperoxide) receptor.

Blood Platelets↗