Interaction Of Plasma Proteins With Artificial Surfaces With Reference To Platelet Adhesion

1981 ◽  
Author(s):  
O P Malhotra ◽  
M N Helmus ◽  
D F Gibbons
Keyword(s):  
Plasma Proteins ◽  
Platelet Adhesion ◽  
Platelet Rich Plasma ◽  
Gel Filtration ◽  
Platelet Suspension ◽  
Platelet Poor Plasma ◽  
Artificial Surfaces ◽  
Glass Surfaces

On occasion, fewer platelets from platelet rich plasma (PRP), adhered to hydrophilic (glass) surfaces exposed to platelet poor plasma (PPP) for 3 min than areas exposed for 3 s. The decrease was dramatic and consistent when platelet suspension (gel-PLS, obtained from PRP by 2B Sepharose gel filtration) was used instead of PRP. To further explore the factors which influence platelet adhesion, we used the following: for surfaces, a) sparkleen-cleaned glass (hydrophilic), b) acid-washed (somewhat hydrophobic), and c) siliconized (hydrophobic); for proteins, a) PPP, b) fibrinogen (96% clottable), c) defi-brinogenated (defib.) plasma, and d) defib. plasma plus fibrinogen; for platelet suspension, a) PRP, b) gel-PLS, and c) platelets in defib. plasma (defib. PLS).From gel-PLS, non-siliconized surfaces exposed to fibrinogen for 3 s attached more platelets (F<0.05) than those exposed to PPP or defib. plasma plus fibrinogen. The latter two attracted more platelets (P<0.01) than defib. plasma. Hydrophilic sparkleen-cleaned glass previously exposed to PPP (or defib. plasma plus fibrinogen) attached a minimum of 10-fold as many platelets from gel-PLS than from PRP. Under similar conditions acid-cleaned surfaces attached 2-fold from gel-PLS, while hydrophobic glass did not show any change. By exposing the surfaces to PPP followed by gel-PLS, the sparkleen-cleaned glass showed the greatest decrease (P<0.001) in the number of platelets attached to areas exposed to PPP for 3 min as compared to 3 s, while siliconized showed no such decrease. If, however, the surfaces were exposed to defib. plasma, they all showed decreases in platelet attraction at 3 min.

Building and Clinical Use of Activated Plasma Clotting Time Test and the Value of APCT on Predicting the Bleeding Due to Leukemia Chemotherapy-Induced Thrombocytopenia.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3915-3915
Author(s):  
Bao-An Chen ◽  
Jin-Jin Li ◽  
Cheng-Yin Huang ◽  
Feng Gao ◽  
Jian Chen ◽  
...  
Keyword(s):  
Sodium Citrate ◽  
Platelet Rich Plasma ◽  
Venous Blood ◽  
Clinical Use ◽  
Clotting Time ◽  
Platelet Suspension ◽  
Platelet Poor Plasma ◽  
Platelet Counts ◽  
Significant Difference ◽  
Time Test

Abstract Objective This study was aimed to determine the optimal concertration of C-PG in activated plasma clotting time (APCT) test and build the APCT test.To investigate the value of activated plasma clotting time test (APCT) on predicting the bleeding due to Leukemia chemotherapy-induced thrombocytopenia. Method We collected healthy blood donors’venous blood, prepared watched platelet suspension and platelet rich plasma in a normal way to which we add different concertrations of C-PG, then test expression of AnnexinV in the surface of activated platelet by FCM. Prepared platelet rich plasma and platelet poor plasma in a normal way to which we add different concertrations of C-PG, then count the plasma clotting time.Forty-one patients with leukemia chemotherapy, twenty-one patients with petechiae or epistaxis or gum bleeding and twenty patients without bleeding symptoms, were involved in the test. Drawed their venous blood and collected in 3.2% buffered sodium citrate(9:1), the platelet counts and APCT, PT, APTT, TT, Fig were assesed. Result Bleeding group’s APCT was obviously prolonged contrast to the non-bleeding group’s. Platelet counts and APCT of the bleeding group and the non-bleeding group were (7.2±2.9) ×109/L, (105.9±8.1)s and (23.0±12.2) ×109/L,(71.4±9.0)s respectively. There was significant difference between the two groups (P&lt;0.01). Using APCT≥90s as the cut-off point to predict the bleeding caused by chemotherapy-induced thrombocytopenia, the sensibility was 100%, and the specificity was 95.7%. Using platelet counts ≤15*109/L, the sensibility and the specificity was 95.2% and 69.0%. PT, APTT, TT, Fig of the bleeding group and the non-bleeding group were 11.7±1.1)S,(31.5±1.3)S,(11.1±1.2)S,(2.37±0.41)g/L and (12.1±0.8)S,(30.8±2.1)S,(10.7±0.9)S,(2.64±0.27)g/L respectively. The results of the two groups were all not significantly different (P&gt;0.05). Conclusion APCT is a good indication of predicting the bleeding due to leukemia chemotherapy -induced thrombocytopenia.

scholarly journals Effect of platelet-activating factor (PAF) on human platelets

Blood ◽  
1982 ◽  
Vol 59 (3) ◽  
pp. 582-585 ◽  
Author(s):  
CM Chesney ◽  
DD Pifer ◽  
LW Byers ◽  
EE Muirhead
Keyword(s):  
Platelet Rich Plasma ◽  
Gel Filtration ◽  
Platelet Activating Factor ◽  
Creatine Phosphate ◽  
Major Effect ◽  
Human Platelets ◽  
Metabolic Inhibitors ◽  
Platelet Factor ◽  
Platelet Suspension ◽  
Second Wave

Abstract The effect of pure synthetic PAF (1–0-alkyl-2-acetyl-sn-glycero-3- phosphorylcholine) was studied in human platelets. PAF (0.2--2.0 micrograms/ml) produced a dose-dependent aggregation in human platelet- rich plasma (PRP) or platelet suspension obtained by gel-filtration (GFP). In addition, PAF (0.8 microgram/ml) induced secretion of 14C- serotonin (45% +/- 10%; mean +/- SD, n = 9) and platelet factor 4 (PF4) (12.89 +/- 3.81 micrograms/10(9) platelets; n = 9) in PRP. Similar results were obtained in GFP. Aggregation and release of 14C-serotonin and PF4 were inhibited by the metabolic inhibitors 2-deoxyglucose (16.7 mM) and antimycin-A (8.3 micrograms/ml), by the membrane-active drugs mepacrine (10 microM) and chlorpromazine (0.025 mM), by PGI2 (5.34 nM), which elevates intracellular c-AMP, by indomethacin (10 microM) or aspirin (100 microM). The ADP scavengers, creatine phosphate and creatine phosphokinase (CP/CPK), inhibited the second wave of aggregation but not secretion. These data suggest that the major effect of PAF on human platelets is mediated through the cyclo-oxygenase pathway and not through a third pathway.

Studies on Platelet-Heparin Interactions Utilizing a Tritium-Labelled Heparin

1975 ◽  
Author(s):  
J. N. Shanberge ◽  
J. Kambayashi
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Surface Charge ◽  
Plasma Protein ◽  
Plasma Proteins ◽  
Liquid Scintillation ◽  
Platelet Rich Plasma ◽  
Scintillation Counter ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Heparin Plasma

Intact platelets were separated from platelet-rich plasma by means of sepharose 2B columns according to the method of Tangen, Eerman and Marfey. 90% of the platelets were recovered, free of plasma proteins. When the platelet-rich plasma was treated with tritium-labelled heparin prior to chromatography the platelets obtained by gel filtration contained radioactivity as measured in a liquid scintillation counter. Radioactivity also was found in the plasma protein fractions which came out of the column before the platelet fractions. However, these latter fractions were eluted earlier than active fractions obtained when tritium-labelled heparin alone was chromatographed. Platelets obtained by gel filtration were treated with tritium-labelled heparin and then rechromatographed. Again platelets with radioactivity were obtained. Treating the platelet-rich plasma with sodium dodecyl sulfate, which alters surface charge, prevents the platelets from taking up heparin while heparin-plasma protein complexing does not seem to be inhibited. Attempts to release heparin from intact platelets has been unsuccessful. Whether platelets combine with heparin by surface complexing or by absorption has not been determined, but the ability of platelets to take up heparin does not seem dependent on prior heparin-plasma protein complexing.

scholarly journals Effect of platelet-activating factor (PAF) on human platelets

Blood ◽  
1982 ◽  
Vol 59 (3) ◽  
pp. 582-585
Author(s):  
CM Chesney ◽  
DD Pifer ◽  
LW Byers ◽  
EE Muirhead
Keyword(s):  
Platelet Rich Plasma ◽  
Gel Filtration ◽  
Platelet Activating Factor ◽  
Creatine Phosphate ◽  
Major Effect ◽  
Human Platelets ◽  
Metabolic Inhibitors ◽  
Platelet Factor ◽  
Platelet Suspension ◽  
Second Wave

The effect of pure synthetic PAF (1–0-alkyl-2-acetyl-sn-glycero-3- phosphorylcholine) was studied in human platelets. PAF (0.2--2.0 micrograms/ml) produced a dose-dependent aggregation in human platelet- rich plasma (PRP) or platelet suspension obtained by gel-filtration (GFP). In addition, PAF (0.8 microgram/ml) induced secretion of 14C- serotonin (45% +/- 10%; mean +/- SD, n = 9) and platelet factor 4 (PF4) (12.89 +/- 3.81 micrograms/10(9) platelets; n = 9) in PRP. Similar results were obtained in GFP. Aggregation and release of 14C-serotonin and PF4 were inhibited by the metabolic inhibitors 2-deoxyglucose (16.7 mM) and antimycin-A (8.3 micrograms/ml), by the membrane-active drugs mepacrine (10 microM) and chlorpromazine (0.025 mM), by PGI2 (5.34 nM), which elevates intracellular c-AMP, by indomethacin (10 microM) or aspirin (100 microM). The ADP scavengers, creatine phosphate and creatine phosphokinase (CP/CPK), inhibited the second wave of aggregation but not secretion. These data suggest that the major effect of PAF on human platelets is mediated through the cyclo-oxygenase pathway and not through a third pathway.

The Effect of Lysed Platelets on Neutralization of Heparin In Vitro with Protamine as Measured by the Activated Coagulation Time (ACT)

1991 ◽  
Vol 66 (02) ◽  
pp. 213-217 ◽  
Author(s):  
Arthur P Bode ◽  
William J Castellani ◽  
Edna D Hodges ◽  
Susan Yelverton
Keyword(s):  
Platelet Rich Plasma ◽  
Coagulation Time ◽  
Platelet Factor ◽  
Platelet Suspension ◽  
Antiheparin Activity ◽  
Unit Increase ◽  
Heparin Anticoagulation ◽  
Heparin Activity ◽  
Activated Coagulation Time

SummaryThe effect of lysed platelets on the activated coagulation time (ACT) was studied in heparinized whole blood during titration with protamine. Frozen-thawed washed platelet suspension, or a chromatography fraction thereof, or autologous frozen-thawed platelet-rich plasma was added in various dilutions to freshly drawn blood anticoagulated with 3,000 USP units/1 heparin. After a 10 min incubation, the amount of protamine needed to restore the ACT to baseline ("protamine titration dose") was determined. We found that the protamine titration dose decreased in proportion to the amount of lysed platelet material added; expressed as a percentage of the total number of platelets present, each unit increase in lysed platelets produced a 1.7% ±0.8 (SD) reduction in the protamine dose needed to normalize the ACT. A heparin activity assay showed that this effect was not due to antiheparin activity of lysed platelets such as platelet factor 4 (PF4). Our data indicate that the procoagulant activity of platelet membranes reduced the sensitivity of the ACT to heparin. These findings suggest that membranous platelet microparticles may cause an inaccurate calculation, based on the ACT, of a protamine dose to reverse heparin anticoagulation in cardiopulmonary bypass procedures.

Purification of the Antihemophilic Factor by Gel Filtration on Agarose

1969 ◽  
Vol 22 (03) ◽  
pp. 577-583 ◽  
Author(s):  
M.M.P Paulssen ◽  
A.C.M.G.B Wouterlood ◽  
H.L.M.A Scheffers
Keyword(s):  
Factor Viii ◽  
Plasma Proteins ◽  
Large Scale ◽  
Gel Filtration ◽  
Large Scale Preparation ◽  
Scale Preparation ◽  
Antihemophilic Factor

SummaryFactor VIII can be isolated from plasma proteins, including fibrinogen by chromatography on agarose. The best results were obtained with Sepharose 6B. Large scale preparation is also possible when cryoprecipitate is separated by chromatography. In most fractions containing factor VIII a turbidity is observed which may be due to the presence of chylomicrons.The purified factor VIII was active in vivo as well as in vitro.

Inhibitory Effect of Staphylokinase on Platelet Aggregation

1993 ◽  
Vol 70 (05) ◽  
pp. 834-837 ◽  
Author(s):  
Akira Suehiro ◽  
Yoshio Oura ◽  
Motoo Ueda ◽  
Eizo Kakishita
Keyword(s):  
Platelet Aggregation ◽  
Human Platelet ◽  
Degradation Products ◽  
Platelet Rich Plasma ◽  
Inhibitory Effect ◽  
Effective Concentration ◽  
Inhibit Platelet Aggregation ◽  
Platelet Suspension ◽  
Washed Platelet ◽  
Human Platelet Aggregation

SummaryWe investigated the effect of staphylokinase (SAK), which has specific thrombolytic properties, on human platelet aggregation. Platelet aggregation induced with collagen was observed following preincubation of platelets in platelet-rich plasma (PRP) or washed platelet suspension (WP) with SAK at 37° C for 30 min. SAK inhibited platelet aggregation in PRP only at the highest examined concentration (1 x 10-4 g/ml). Although SAK did not inhibit platelet aggregation in WP which contained fibrinogen, it did when the platelets had been preincubated with SAK and plasminogen. The most effective concentration in WP was 1 x 10-6 g/ml. The effect could be inhibited by adding aprotinin or α2-antiplasmin. The highest generation of plasmin in the same preincubation fluid was detected at 1 x 10-6 g/ml SAK. We concluded that SAK can inhibit platelet aggregation in WP by generating plasmin and/or fibrinogen degradation products, but is only partially effective in PRP because of the existence of α2-antiplasmin.

Antiheparin Activity of Human Serum and Platelet Factor 4

1973 ◽  
Vol 30 (01) ◽  
pp. 093-105 ◽  
Author(s):  
C.H.J Sear ◽  
L Poller ◽  
F.R.C Path
Keyword(s):  
Molecular Weight ◽  
Ionic Strength ◽  
Blood Serum ◽  
Whole Blood ◽  
Platelet Rich Plasma ◽  
Gel Filtration ◽  
Normal Serum ◽  
Platelet Factor ◽  
Mean Values ◽  
Antiheparin Activity

SummaryThe antiheparin activity of normal serum has been studied by comparing the antiheparin activities of sera obtained from normal whole blood, platelet-rich plasma and platelet-’free’ plasma with a purified platelet extract during differential isoelectric precipitation and by gel filtration chromatography.The mean values for the activity of PRP-serum and PFP-serum were 106% (S.D. 11) and 10% (S.D. 3) of untreated whole blood respectively. The activity of whole blood serum, PRP serum and whole blood serum plus platelet extract precipitated under identical physical conditions, i.e. pH 7.0, I =0.008, indicating that the activities of the three samples are probably associated with PF4. PF4 precipitated from human platelet extract at pH 4.0, but this is probably due to the difference in the two biochemical environments investigated, i.e. serum and platelet extract.The gel filtration experiments revealed striking similarities between the major antiheparin activities of serum and platelet extract. At physiological pH and ionic strength both activities were associated with high molecular weight material, but at physiological pH and elevated ionic strength both activities behaved as much smaller entities of molecular weight between 25,000 and 30,000 daltons and it seems very likely that both activities are associated with the same molecule, i.e. PF4.

Soluble Fibrin Complexes and Fibrinogen Heterogeneity in Diabetes mellitus

1980 ◽  
Vol 44 (03) ◽  
pp. 130-134 ◽  
Author(s):  
E B Tsianos ◽  
N E Stathakis
Keyword(s):  
Diabetes Mellitus ◽  
Molecular Weight ◽  
Plasma Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Lower Molecular Weight ◽  
Polyacrylamide Gels ◽  
Soluble Fibrin ◽  
Α2 Macroglobulin ◽  
Patients With Diabetes

SummaryThe presence of soluble fibrin complexes (SFC) measured by gel filtration of plasma on 4% agarose columns, fibrinogen heterogeneity on 3.5% SDS-polyacrylamide gels and the concentrations of several plasma proteins were evaluated in 39 patients with diabetes mellitus (DM) and 19 matched control subjects. A small but significant increase of SFC was found in DM (p<0.01). On individual basis 51.2% of the patients had increased SFC (>M + 2 SD of the controls). Polyacrylamide gel electrophoresis of the SFC showed no evidence of cross-linking or proteolysis. Plasma clots formed in the presence of EDTA and trasylol were analysed in SDS-polyacrylamide gels in a normal and two lower molecular weight fibrin bands (band I, II, III). The percentage of band I fibrinogen was in diabetics (65.3 ± 4.7%) lower than that of the controls (71.8 ± 4.5%) (p < 0.01). Fibrinogen levels, antithrombin III, α1-antitrypsin, α2-macroglobulin and plasminogen were significantly increased in DM. We suggest that in DM there is an enhancement of intravascular fibrin formation and accelerated fibrinogen degradation to lower molecular weight forms.

Baboon Fibrinogen Adsorption and Platelet Adhesion to Polymeric Materials

1991 ◽  
Vol 65 (05) ◽  
pp. 608-617 ◽  
Author(s):  
Joseph A Chinn ◽  
Thomas A Horbett ◽  
Buddy D Ratner
Keyword(s):  
Platelet Adhesion ◽  
Coagulation Factor ◽  
Polymeric Materials ◽  
Platelet Suspension ◽  
Perfusion Chamber ◽  
Static Conditions ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Fibrinogen Adsorption

SummaryThe role of fibrinogen in mediating platelet adhesion to polymers exposed to blood plasma was studied by comparison of the effect of plasma dilution on fibrinogen adsorption and platelet adhesion, and by the use of coagulation factor deficient plasmas. Polyetherurethane substrates were first preadsorbed with dilute plasma, then contacted with washed platelets suspended in a modified, apyrase containing Tyrode’s buffer. Platelet adhesion was studied under static conditions in Multiwell dishes, and also under shearing conditions using a parallel plate perfusion chamber. Fibrinogen adsorption and platelet adhesion were measured using 125I radiolabeled baboon fibrinogen and min radiolabeled baboon platelets, respectively. Surfaces were characterized by electron spectroscopy for chemical analysis (ESCA).When fibrinogen adsorption to Biomer was measured after 2 h contact with a series of dilute plasma solutions under static conditions, a peak in adsorption was observed from 0.26% plasma, i.e., adsorption was greater from 0.26% plasma than from either more or less dilute plasma. A peak in subsequent platelet adhesion to the plasma preadsorbed surfaces, measured after 2 h static incubation with washed platelets, was also observed but occurred on Biomer preadsorbed with 1.0% plasma.When fibrinogen adsorption was measured after 5 min contact under shearing conditions, the fibrinogen adsorption peak occurred on surfaces that had been exposed to 1.0% plasma. A peak in platelet adhesion to these preadsorbed surfaces, measured after 5 min contact with the platelet suspensions under shearing conditions, was observed on Biomer preadsorbed with 0.1% plasma. Shifts between the positions of the peaks in protein adsorption and platelet adhesion occurred on other polymers tested as well.Platelet adhesion was almost completely inhibited when baboon and human plasmas lacking fibrinogen (i. e., serum, heat defibrinogenated plasma, and congenitally afibrinogénémie plasma) were used. Platelet adhesion was restored to near normal when exogenous fibrinogen was added to fibrinogen deficient plasmas. Adhesion was also inhibited completely when a monoclonal antibody directed against the glycoprotein IIb/IIIa complex was added to the platelet suspension. Platelet adhesion to surfaces preadsorbed to von Willebrand factor deficient plasma was the same as to surfaces preadsorbed with normal plasma.While it appears that surface bound fibrinogen does mediate the initial attachment of platelets to Biomer, the observation that the fibrinogen adsorption and platelet adhesion maxima do not coincide exactly also suggests that the degree of subsequent platelet adhesion is dictated not only by the amount of surface bound fibrinogen but also by its conformation.

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