Kinetics of In-111-platelets in the baboon: II. In vitro distribution and sites of sequestration

The kinetics and sites of sequestration of a fully representative population of In-111-platelets were determined in 11 baboons. The in vivo method of quantification with computer assisted scintillation camera image analysis was validated by sacrificing 5 baboons and measuring and comparing the distribution of organ radioactivity. Recovery of platelets in the circulation was 87% +/- 7, and their mean survival time was 147 hr +/- 15. The mean splenic platelet pool was 16.0 +/- 1.9. At equilibrium 15.8% +/- 2.9 of the In-111-platelets were in the hepatic blood pool. Senescent platelets were destroyed in the reticulo-endothelial system. The major sites of sequestration were: liver (37.6% +/- 6.0), and the spleen (23.3% +/- 4.6). The bone marrow sequestrated 14.4% +/- 1.7 of the labelled platelets, and 15.5% +/- 4.0 were present in various other tissues. We conclude that the in vivo method of In-111-quantification is accurate. Senescent platelets are mainly sequestrated in the reticuloendothelial tissue, with the liver, spleen and the bone marrow important sites of sequestration.     

Intrahepatic kinetics of indium-111-labelled platelets

The intrahepatic kinetics of 111indium-labelled platelets have been studied using dynamic gamma camera scintigraphy immediately following injection. Platelets labelled in saline with 111In-oxine or 111In-acetylacetonate underwent rapidly reversible hepatic sequestration, indicating that they were "activated". Platelets labelled in plasma with 111In-tropolonate, however, did not display this phenomenon. On the assumption that plasma-labelled platelets display a normal initial bio-distribution, mean intrahepatic platelet transit time, as a factor of the transit time of 99m-Tc labelled red cells, was 1.45 +/- SE 0.12 (n = 6), implying the normal presence of a small intrahepatic platelet pool. Unlike the liver, transit through the spleen was not sensitive to the labelling medium; thus the mean intrasplenic transit time of plasma-labelled platelets was 9.3 +/- SE 0.7 min (n = 10), and of saline-labelled platelets 9.5 +/- SE 0.3 min (n = 8).     

Platelet kinetics and scintigraphic imaging in thrombocytopenic malaria patients

Thrombocytopenia is a common occurrence in acute malaria. It is attributed, among other factors, to excessive splenic platelet pooling and a shortened platelet lifespan. The aim of our study was to evaluate the platelet kinetics and sequestration site by isotopic studies in uncomplicated malaria-induced thrombocytopenia. Seven thrombocytopenic malaria patients (74,000+/-36,000 platelets/ micro l) were included in the study. Autologous (111)In-labeled platelet scintigraphy was performed up to 96 hours (h) post injection (p.i.) to evaluate the platelet sequestration site. Late sequestration for the spleen (S) and the liver (L) was analyzed according to the following activity ratios: S (spleen count on the last day of the platelet lifespan / spleen count at 30 min) and L (liver count on the last day of the platelet lifespan / liver count at 30 min). Additionally, platelet survival studies were performed. A normal late sequestration (S: 0.95+/-0.06 and L: 1.04+/-0.08; normal values, S and L: 1+/-0.2.) was observed in all of our patients. The platelet lifespan was reduced (1 to 4 days; normal range, 7-9 days), recovery was normal (mean, 63+/-6%; normal range, 55-75%), and the turnover rate was enhanced (mean, 95,000+/-80,000/ micro l/day; normal value, 35,000+/-4,500/ micro l/ day). According to the results of scintigraphy, the sequestration site by uncomplicated malaria-induced thrombocytopenia appears to be non-splenic and/or hepatic, yet diffuse.     

Effects of fish oil supplementation on platelet survival and ex vivo platelet function in hypercholesterolemic patients

Little is known about the effects of dietary supplementation on platelet survival with low doses of n-3 and n-6 fatty acids in patients with hypercholesterolemia. The effects of a 6-week intervention with fish oil capsules (daily intake: 216 mg eicosapentaenoic acid, 140 mg docosahexaenoic acid, 390 mg gamma-linolenic acid, and 3480 mg linoleic acid) on in vivo platelet survival (111 In-oxine labeled platelets) and on ex vivo markers of platelet activation were investigated in a placebo-controlled, double-blind study with 26 hypercholesterolemic patients. In vivo platelet survival increased in the fish oil group (T) from a mean of 159+/-14 hours to a mean of 164+/-12 hours (p=0.025), whereas it remained unchanged in the placebo (P) group (T vs. P; p=0.055). Ex vivo, thromboxane B2 decreased from a mean of 225+/-16 to 212+/-21 ng/mL (p=0.003) in T but did not change in P (T vs. P: p=0.002). Malondialdehyde formation was lowered significantly by fish oil supplementation from a mean of 5.49+/-1.3 to 5.12+/-1.05 nM/10(9) platelets, p=0.005, as compared with P (T vs. P; p=0.018). The trendwise decrease in 11-DH-thromboxane B2 plasma levels was not significant nor was the increase in platelet sensitivity to prostaglandin I2 by fish oil. Baseline platelet survival in patients with hyperlipoproteinemia type IIa was not different from those with hyperlipoproteinemia IIb and response to treatment in terms of platelet activation markers was not either. The changes in platelet activation parameters in T were associated with significant reductions in cholesterol (-2.9%), low density lipoprotein cholesterol (-3.5%), and triglycerides (-12.4%). Both ex vivo and in vivo platelet activation parameters exhibited signs of decreased activation by a 6-week diet supplemented with n-3 and n-6 fatty acids, which might be beneficial in reducing atherothrombotic risk, in patients with hyperlipoproteinemia type IIa and IIb.     

Effects of a monoclonal antibody to glycoprotein IIb/IIIa (P256) and of enzymically derived fragments of P256 on human platelets

The anti-platelet monoclonal antibody P256 is currently undergoing development for in vivo detection of thrombus. We have examined the actions of P256 and two fragments on human platelet function. P256, and its divalent fragment, caused aggregation at concentrations of 10(-9)-3 x 10(-8) M. A monovalent fragment of P256 did not cause aggregation at concentrations up to 10(-7) M. P256-induced platelet aggregation was dependent upon extracellular calcium ions as assessed by quin2 fluorescence. Indomethacin partially inhibited platelet aggregation and completely inhibited intracellular calcium mobilisation. Apyrase caused partial inhibition of aggregation. Aggregation induced by the divalent fragment was dependent upon fibrinogen and was inhibited by prostacyclin. Aggregation induced by the whole antibody was only partially dependent upon fibrinogen, but was also inhibited by prostacyclin. P256 whole antibody was shown, by flow cytometry, to induce fibrinogen binding to indomethacin treated platelets. Monovalent P256 was shown to be a specific antagonist for aggregation induced by the divalent forms. In-111-labelled monovalent fragment bound to gel-filtered platelets in a saturable and displaceable manner. Monovalent P256 represents a safer form for in vivo applications.     

Validation of the dual radiotracer method for quantitative In-111 platelet scintigraphy

We have developed a simple in vivo scintigraphic technique that permits accurate quantitative comparison of intravascular platelet deposition in blood vessels of similar size. Regional count information from scintigraphic images of In-111 platelets and Tc-99m red blood cells is used to determine the ratio of radioactivity in the thrombus relative to that in the blood. Multiplication of this ratio by the percent injected dose (%ID) of In-111 per ml of blood yields a quantity (%ID index) that is directly proportional to the %ID of In-111 platelets in the thrombus as measured in vitro (r = .879, p less than 0.005). This technique is well suited for quantitative studies of scintigraphically detectable platelet deposition in both animals and in man.     

Platelet alpha granule depletion: findings in patients with prosthetic heart valves and following cardiopulmonary bypass surgery

The alpha granule content of platelets, as indicated by the amount of beta-thromboglobulin (beta-TG) in lysed platelet rich plasma was measured to determine whether platelet stimulation resulted in a circulating population of granule-depleted platelets. In 101 normal controls, with a mean platelet count of 242.6 +/- 6.5 X 10(9)/1, the mean platelet beta-TG content was 55.9 +/- 1.2 ng/10(6) platelets. There was a significant reduction in both these parameters (mean platelet count 195.5 +/- 5.8 X 10(9)/1 (P less than 0.001), mean platelet beta-TG 50.0 +/- 1.2 ng/10(6) platelets (P less than 0.01)) in 74 patients with prosthetic cardiac valves. In 24 patients, cardiopulmonary bypass surgery caused a much greater reduction in median platelet count from 210 X 10(9)/1 to 11.1 X 10(9)/1, two hours after surgery (P less than 0.001) but no overall change in platelet beta-TG. However, five patients who experienced diffuse haemorrhage in the postoperative period had a lower median platelet beta-TG (35.5 ng/10(6) platelets) than the other 19 patients (51.0 ng/10(6) platelets) (P less than 0.05).     

Platelet deposition at carotid endarterectomy sites in humans

Following carotid endarterectomy, early postoperative thrombosis or late restenosis occurs in up to 20% of vessels. Both complications may be related to platelet mechanisms. To assess platelet deposition at endarterectomy sites, we injected indium-111 labelled platelets in 24 men less than 30 minutes after carotid endarterectomy, with subsequent imaging 24-96 hours later. To determine if deposition decreased over time, 12 patients had follow-up studies 0.5-24 months later. For comparison, 2 control groups were studied: 1) patients with noncarotid surgery (n = 6) and 2) normal young subjects without endarterectomy and without evidence of carotid disease (n = 12). Quantitative analysis was performed performed using a deposition index that compared activity in operated with unoperated sites in surgical patients or activity in the right with left carotid arteries in normal subjects. Patients with recent endarterectomy had a mean deposition index of 1.7 +/- 0.5 (range 1.2-3.5) compared with a similarly determined ratio of 1.1 +/- 0.1 in normal subjects and 1.2 +/- 0.1 in the surgical controls (both p less than or equal to 0.05 vs. acute endarterectomy). At follow-up after endarterectomy, the mean deposition index decreased to 1.0 +/- 0.1, documenting reduced platelet deposition over time. We conclude that the arterial injury of carotid endarterectomy results in early platelet deposition, which is no longer present in most patients who are studied late. These findings suggest a reduction in platelet thrombus formation with time and are compatible with reendothelialization of the endarterectomized surface. This model may be useful for the in vivo assessment of therapies designed to reduce platelet accumulation following endothelial injury in humans.     

In vivo labeling of platelets with 75Se -- selenomethionine in patients with hepatic cirrhosis and thrombocytopenia

Labeling of platelets in vivo by 75Se -- Selenomethionine (75Se-M) was performed in nine cases of hepatic cirrhosis and thrombocytopenia for evaluation of the kinetics of platelet maturation. Folic acid and vitamin B12 deficiency was excluded by pretreatment of the patients with these agents. The platelet maturation time -- time between the injection of the isotope and maximum radioactivity of separated blood platelets -- was shortened to 7.7 +/- 1.1 days (mean +/- SD) compared to the normal 9.1 +/- 1.4 days. For explanation a disturbance of megakaryocyte maturation and/or platelet release from the bone marrow is suggested.     

Platelet production, clearance and distribution in patients with idiopathic thrombocytopenic purpura

Platelet destruction mechanism was thought to play a primary role in autoimmune thrombocytopenia (ITP). There is, however, some evidence that anti-platelet antibodies in ITP impair megakaryocytopoiesis. Using autologous In-111 platelets, we tried to elucidate this point. We measured platelet survival, platelet turnover, platelet sequestration sites, and platelet production (turnover) to the clearance (sum of liver and spleen platelet uptake) ratio in 8 normal subjects and 12 patients with ITP whose platelet counts ranged from 9 x 10(9) to 40 x 10(9)/L. The sum of platelet uptake in the liver and spleen showed a significant inverse correlation with platelet survival. Platelet survival, platelet production to clearance ratio correlated significantly with the platelet count. No significant correlation was found between platelet turnover and platelet counts. The distribution of platelet turnover showed considerable individual variation; 8 of 12 patients showed platelet turnovers which were lower than the normal value, but the others were within the normal range. We concluded that although the platelet destruction mechanism in the reticuloendothelial system shows a primary role in thrombocytopenia, the impaired rate of effective thrombopoiesis may also contribute to the severity of ITP.     

The effect of hypoxia on platelet survival and site of sequestration in the newborn rabbit

Thrombocytopenia occurs frequently in sick neonates that have experienced perinatal asphyxia. This study investigated the effect of one component of asphyxia, hypoxia, on platelet lifespan and site of sequestration. 111Indium oxine platelet survivals with scintigraphic imaging were performed in newborn and adult rabbits exposed to room air (normoxia) or following exposure to a 15 minute, severe hypoxic insult (FIO2 = 0.05). Platelet survivals in normoxic adults (n = 27) and newborn rabbits (n = 11) were similar (60 +/- 3.9 hr vs 64 +/- 8.0 hr, m +/- SEM). Inhalation of 5% oxygen for 15 minutes was not associated with an acidemia and did not produce thrombocytopenia but significantly shortened the platelet survival to 34 +/- 3 hr in the adult (n = 18) and 38 +/- 3 hr in the newborn rabbit (n = 7). Postmortem measurement of the sites of 111In-platelet accumulation showed that under normoxic conditions the platelets accumulated in the liver and spleen (23 +/- 4.3% and 8 +/- 1.0% of the total body counts) in the adult with even greater accumulation in the liver (58 +/- 6.8%) and spleen (19 +/- 4.9%) of the newborn (p less than 0.001). The latter observation was likely due to the relatively increased size of the liver and spleen in the newborn compared to the adult. Hypoxia did not alter the site of platelet sequestration in adults or newborns. Our results suggest that the newborn has the same platelet survival as the adult and that acute, severe hypoxia significantly shortens the survival of platelets in both groups. Although the sites of sequestration are qualitatively the same in the newborn, there is greater sequestration in the liver and spleen when compared to the adult.     

Kinetics of In-111-platelets in the baboon: I. Isolation and labelling of a viable and representative platelet population

A fully representative and viable platelet population was isolated from the blood of 15 baboons by a multiwash procedure, and labelled with In-111-oxine. The recovery of the total platelet population in the circulation was 85% +/- 9. Mean platelet life span was 146 hr +/- 13. Correcting for plasma radioactivity (always less than 3.5%) did not significantly affect the estimate of platelet life span (145 hr +/- 16) or recovery (85% +/- 12). Platelet survival estimates, repeated at different times, were reproducible. In 5 baboons, platelets were also harvested by a single step differential centrifugation. The mean life span of a representative platelet population was significantly longer than that of platelets harvested by a single step. Recovery values of the representative and non-representative population were similar. We conclude that it may be important to harvest and label a fully representative platelet population for kinetic studies. The proposed method is simple and reproducible, and may be applied in studies in humans.     

Platelet membrane phospholipids in euthymic bipolar disorder patients: are they affected by lithium treatment?

BACKGROUND: Abnormalities in cell membrane processes and intracellular signal transduction pathways may be implicated in the pathophysiology of bipolar disorder. In this study, we attempted to investigate, in euthymic bipolar patients: 1) in vivo signal transduction abnormalities of the phosphatidylinositol pathway in platelets; and 2) possible in vivo effects of lithium treatment on platelet membrane phospholipids. METHODS: We determined the relative absorbances of eight individual classes of platelet membrane phospholipids, using two-dimensional thin-layer chromatography in high-performance plates, followed by scanning laser densitometry, in a group of 10 lithium-treated euthymic bipolar patients and 11 normal controls. RESULTS: The mean relative absorbance of phosphatidyl-inositol-4,5-bisphosphate (PIP2) was lower in the patient group (0.29 +/- 0.08% vs. 0.39 +/- 0.12%; t = 2.35, df = 19, p = .03); no significant differences between patients and controls were found for the other phospholipids. CONCLUSIONS: This study provides in vivo evidence that bipolar patients on lithium treatment exhibit a decreased relative amount of PIP2 in the platelet cell membranes compared to normal controls.     

Autologous platelet-labeling in thrombocytopenia

Field studies performed with peripheral platelets obtained from 6 male volunteers aged 23 to 29 years revealed an extraordinary dependence of labeling efficiency on incubation time and platelet concentration after 111In-oxine platelet labeling. Since the monitoring of in vivo-platelet function in patients with thrombocytopenia may cause problems due to insufficient labeling results and homologous platelets may show a different in vivo behaviour to autologous ones, we have searched for the minimal amount of platelets necessary to allow appropriate labeling and imaging in patients with thrombocytopenia. In 15 patients with untreated thrombocytopenia aged 14 to 79 years demonstrating a mean peripheral platelet count of 2.509 +/- 1.45 x 10(4) cells/microliters autologous 111In-oxine platelet labeling was performed. The results indicate that approximately 1 x 10(8) (concentrated) platelets/ml are necessary to obtain an adequate labeling efficiency and recovery. This platelet concentration can be easily achieved by drawing one more Monovette of whole blood per each 5 x 10(4) platelets/microliter peripheral platelet count less than 2 x 10(5)/microliter. It is concluded, that calculation of the required number of platelets in advance, variation of the blood volume drawn and the volume of incubation buffer allow informative, qualitative and quantitative results using autologous platelets. The method presented effectively circumvents the requirement of homologous platelets for radiolabeling in thrombocytopenia.     

Indium-111-labelled human platelets: a method for use in severe thrombocytopenia

We describe and evaluate a simple method for labelling autologous human platelets with Indium-111-oxine in patients with severe thrombocytopenia. Twenty patients with immune thrombocytopenia and platelet counts ranging from 5 to 119 X 10(9)/1 were investigated. Platelets were isolated from blood by differential centrifugation, residual platelets were repeatedly washed from the red cell layer and buffy coat and labelled with In 111 in saline. A mean of 55% +/- 21 of platelets were harvested from the blood, labelled with 49% +/- 24 efficiency and 15.8 X 10(8) labelled platelets reinjected to the patients. Contamination of the platelets with red cells and plasma was low. The labelled platelets were viable as assessed by in vitro aggregation, recovery in the circulation and mean survival time. This method permits quantitative platelet imaging with autologous labelled platelets in patients with severe thrombocytopenia.     

Indium-111 labeled platelets: Studies on preparation and evaluation of in vitro and in vivo functions

Platelets have been labeled with a lipid soluble complex of the radionuclide indium-111. The complex is formed with 8-hydroxyquinoline and extracted in chloroform which is then evaporated to dryness. The residue is dissolved in 50 μl of ethanol and diluted to 200 μl with normal saline. Platelets are separated by differential centrifugation, washed and suspended either in Tyrodes-albumin solution or in normal saline. The solution of ¹¹¹In-complex is added to separated platelets and more than 95% of the radioactivity is incorporated with the platelets. The function of the labeled platelets has been studied by their aggregability using adenosine diphosphate and collagen as the stimulating agents. No adverse effects have been observed. The survival of the indium-labeled platelets was similar to that observed with chromium-51 labeled platelets. Labeled canine platelets have been administered to dogs in whom venous thrombi had been induced by alteration of the intima by an electric current. The thrombi were detected by imaging 50 minutes after administration of the labeled platelets. Twenty-four hours later the thrombi were removed and had 20–50 times the radioactivity of an equal weight of blood. Accumulation of large amounts of radioactivity in surgical wounds has also been observed. Damage of the intima of carotid arteries in animals by a balloon catheter demonstrated 8 to 20 times more activity in the damaged artery than in the normal artery. Results indicate that In-111 labeled platelets have potential both for platelet survival studies and for evaluation of vascular damage.     

Quantitation of platelet glycoprotein IV (CD36) in healthy subjects and in patients with essential thrombocythemia using an immunocapture assay.

Glycoprotein IV (GPIIb, CD36) is a major platelet membrane glycoprotein which is thought to participate in a number of adhesive reactions and to mediate signal transduction. In order to measure the total content of GPIV in human platelets, we have developed a simple and sensitive solid-phase radioimmunoassay based on the immunocapture of GPIV from Triton X-100-solubilized platelets. FA6-152, a monoclonal antibody to GPIV was coated on microtiter plates and bound antigen was quantified with a radiolabeled polyclonal antibody to GPIV. Using purified GPIV as a standard, the coefficients of variation of the assay were found to be less than 10% at concentrations of GPIV ranging from 0.15 to 0.75 micrograms/ml. The assay was validated by the parallelism obtained between purified GPIV dose-response curves and those obtained with platelet lysates, indicating a similar antigenic activity for GPIV in both samples. The level of GPIV in platelets from healthy donors was 0.23 +/- 0.05 (mean +/- SD, n = 15) micrograms per 100 micrograms of platelet proteins and a mean value of 27,440 +/- 6,200 (SD) molecules per platelet was calculated. The radioimmunoassay could be used to discriminate between the high level of platelet GPIV in patients with essential thrombocythemia (mean +/- SD = 81,850 +/- 27,780 molecules/platelet; n = 8) and the normal GPIV level in patients with secondary thrombocytosis (mean +/- SD = 26,810 +/- 4,030 molecules/platelet; n = 5), thereby demonstrating the clinical usefulness of the assay. The specific increase in platelet GPIV in patients with essential thrombocythemia was confirmed by immunoblot analysis whereas no increase in platelet GPIb or GPIIb-IIIa was observed by this technique.     

Effect of aspirin and ticlopidine on platelet deposition in carotid atherosclerosis: assessment by indium-111 platelet scintigraphy

The antiplatelet effects of aspirin and ticlopidine were studied by a dual-tracer method, using indium-111 labeled platelets and technetium-99m human serum albumin, in a group of 12 patients with suspected ischemic cerebrovascular disease. The magnitude of platelet accumulation at the carotid bifurcation was expressed as the ratio of radioactivity of indium-111 platelets deposited on the vascular wall to those circulating in the blood-pool (PAI, platelet accumulation index), 48 hr after injection of labeled platelets. PAI values were measured before (baseline studies) and after the antithrombotic therapies (aspirin studies: 325 mg bid for 22.3 +/- 1.3 days, ticlopidine studies: 100 mg tid for 21.8 +/- 2.1 days). At the baseline, the mean PAI value at 24 carotid bifurcations in the patient group was 15.7 +/- 15.3% (mean +/- S.D.) compared to -4.3 +/- 9.1 at 24 carotid bifurcations in 12 normal subjects (p less than 0.01). We defined the upper limit for a normal PAI (%) value to be +13.9, namely the mean PAI plus 2 SD for the carotid bifurcation in normal subjects and used this value for semiquantitative analysis. At the baseline, significant elevation of PAI (more than 13.9%; positive scintigram) was observed at 12 of 24 vessels, while 12 other regions were negative (less than 13.9%). In the lesions with positive scintigraphic results at the baseline, the mean PAI (%) value from the baseline, aspirin and ticlopidine studies was 29.5 +/- 7.0, 11.2 +/- 8.5 (p less than 0.01 versus baseline) and 21.4 +/- 21.3 (not significant from baseline), respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

In vitro thromboxane synthesis of depleted blood platelets following renal transplantation

Renal transplant rejection is associated with platelet activation in vivo which may lead to partially alpha- and delta-granule-depleted platelets that continue to circulate. These "exhausted" platelets are hemostatically defective. To quantitate the extent of platelet granule depletion following kidney transplantation, we determined intraplatelet levels of beta-thromboglobulin (beta TG), platelet factor 4 (PF4), and serotonin (5-hydroxytryptamine, 5-HT) ex vivo in Triton X-100-treated platelet lysates. To explore biochemical alterations of partially depleted platelets, we studied platelet thromboxane A2 (TXA2) synthesis in citrated platelet-rich plasma (PRP) upon stimulation with thrombin or collagen in 45 recipients of renal allografts and 10 healthy volunteers. The patients were divided into subjects with acute and chronic allograft rejection (N = 15), those with compensated renal failure after kidney transplantation but without evidence of allograft rejection (N = 15), and those with functioning renal transplant (N = 15). The mean intraplatelet content of beta TG (38.6 +/- 4.2 micrograms/10(9) platelets), PF4 (11.8 +/- 1.8 micrograms/10(9) platelets), and 5-HT (274 +/- 31 ng/10(9) platelets) in patients with acute or chronic renal allograft rejection was significantly lower than in other recipients of kidney transplants or healthy volunteers (beta TG: 59.9 +/- 4.7 micrograms/10(9) platelets; PF4: 20.4 +/- 2.3 micrograms/10(9) platelets; 5-HT: 461 +/- 48 ng/10(9) platelets; p less than 0.005 in all cases).(ABSTRACT TRUNCATED AT 250 WORDS)     

Platelet inhibitory drugs: an in vivo method of evaluation in patients

The measurement of platelet deposition in human thrombi is essential for the evaluation of platelet-inhibitory drugs and prosthetic materials for use in patients. The rate of 111Indium-labelled platelet accumulation on Dacron arterial grafts was measured in 27 patients randomised to take either aspirin and dipyridamole (ASA + DPM) or placebo. Autologous platelets were labelled and re-injected seven days following surgery and the graft thrombogenicity index calculated as the daily rise in the ratio of emissions from the graft over a reference site. The mean (+/- SD) thrombogenicity index in 12 patients undergoing femoro-popliteal bypass was 0.25 +/- 0.09 on placebo and 0.16 +/- 0.07 on ASA + DPM started pre-operatively (p less than 0.05). Post-operative ASA + DPM therapy started two days following platelet labelling in 15 patients with aorto-femoral grafts also significantly reduce thrombogenicity to 0.12 +/- 0.05 compared with 0.25 +/- 0.08 on placebo (p less than 0.01). In the latter patients the ratio of emissions from the graft over reference fell significantly on starting ASA + DPM, suggesting a net loss of platelets from the graft. These results indicate that the rate of in vivo platelet accumulation on Dacron grafts can be quantitated and that ASA + DPM reduced this rate in man.