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Biomedical subjects

L Poller

Publications and source records attributed to L Poller.

At least 55 records · Page 3Linked to original sources

The diagnosis of lupus anticoagulants by the activated partial thromboplastin time--the central role of phosphatidyl serine.

Liposomes of pure phospholipids were used in a modified APTT test system and the role of phosphatidyl serine (PS) in determining the sensitivity of the test system to the presence of lupus anticoagulants was assessed. Six consecutive patients with lupus anticoagulants and seven haemophiliacs with anticoagulants directed at specific coagulation factors, were studied. Increasing the concentration of phospholipid in the test system markedly reduced the sensitivity to lupus anticoagulants but had marginal effect on the specific factor inhibitors. The same effect was achieved when the content of PS alone was increased in a vehicle liposome of constant composition. The results suggest that the lupus anticoagulants can best be detected by a screening method using an APTT test with a reagent of low PS content. The use of a reagent rich in PS will largely abolish the lupus anticoagulant's effect on the APTT. An approach using the two different types of reagent may facilitate differentiation of lupus inhibitors from other types of anticoagulant.

Adult

The bleeding time: current practice in the UK.

A questionnaire survey of current practice in the bleeding time test has been undertaken by the UK External Quality Assessment Scheme in blood coagulation. Completed returns have been received from 358 centres. Most centres (88.5%) perform bleeding times and of these the Ivy test is the most commonly performed. Only 13.6% perform the Duke method. Templates are used to control the procedure by approximately half of the hospitals. There is considerable variability in the type and depth of incision and interpretation of the endpoint. The upper limit of normality not unexpectedly differs considerably between the centres with both Ivy and Duke methods. The use of a commercial template method, 'Simplate', provides a measure of agreement amongst the group of hospitals using this instrument but it remains to be established whether this is the most reliable procedure. In the interim, gross discrepancies in technique or interpretation should be corrected in the light of the findings of the survey.

Bleeding Time

Lipid class composition and heparin sensitivity in the activated partial thromboplastin time.

In an APTT reagent, prepared from purified lipids, the role of phosphatidyl serine (PS) in determining the sensitivity of the APTT test system to measurement of the effect of heparin in plasma has been evaluated. As the concentration of PS decreases sensitivity to heparin increases but procoagulant activity decreases. Dilution of the test liposome over a wide range (1 g/l to 30 mg/l) had a minimal effect on the clotting time. At levels below 30 mg/l, however, the amount of total lipid appeared to be rate limiting; a loss of procoagulant activity being paralleled by an increase in heparin sensitivity. Phosphatidyl inositol (PI) was not a satisfactory substitute for PS in the APTT method studied. The degree of unsaturation of test liposomes appeared to have no effect on either procoagulant activity or sensitivity to heparin at the lipid concentration employed. In the light of these findings, a more critical appraisal of the phospholipid components of APTT reagents should facilitate the development of more reliable reagents for heparin control. A further benefit of this type of approach should be a reduction in the acknowledged wide variations in sensitivity to heparin which exist between available APTT reagents.

Blood Coagulation Tests

The procoagulant activity of partial thromboplastin extracts: the role of phosphatidyl serine.

The role of phosphatidyl serine (PS) in the procoagulant activity of the APTT test has been studied further, using extracts of improved purity derived from a combination of high performance liquid chromatography and thin layer chromatography. The central role of PS in the APTT has been confirmed but the anticoagulant effect described in our previous report has been localised to contamination with phosphatidyl inositol (PI). A comparatively weak procoagulant activity was detected in purified PI- and lyso PS-containing liposomes and the former had an inhibitory action in mixtures diluted to less than 1 microgram/litre. This was enhanced by the presence of PS in combination experiments. Some correlation was noted between the degree of unsaturation of test liposomes and their procoagulant activity. The observations suggest that PS has a precise and unique role in the procoagulant activity expressed in this test system. PS possesses a strong negative charge and has two fatty acid groups. Other negatively charged phospholipids in this test system were less active. It appears, therefore, that the packing of PS into the lipid bilayer, which depends upon the length and degree of unsaturation of its fatty acid groups, is of considerable importance in establishing the surface topography essential for optimal procoagulant activity. Knowledge of the role played by various phospholipids in the procoagulant activity of the APTT reagent should enable improved standardisation and quality control in the future.

Blood Coagulation

An evaluation of APTT monitoring of low-dose heparin dosage in hip surgery.

The activated partial thromboplastin time has been used to monitor the effects of low-dose subcutaneous heparin in two groups of patients undergoing hip surgery. The study was performed to determine the degree of anticoagulation required to protect these high-risk patients from post-operative deep vein thrombosis. The patients were randomised to receive a fixed regimen of subcutaneous calcium heparin (5,000 units eight-hourly) or a dose of calcium heparin monitored by maintaining the standardised APTT at 50 secs. In the adjusted group the APTT achieved the target figure in 46% of observations compared to 27% in the fixed group (p less than 0.005). Nine patients showed positive 125I-fibrinogen scans and in all, the APTT was below the target value the day before the scan became positive. In contrast, in six of the nine thrombotic patients heparin was detected by antifactor Xa clotting assay. The APTT, therefore, appears to give a better guide to the antithrombotic effect of heparin than the antifactor Xa clotting assay. These preliminary observations suggest that prolonging the standardised APTT method to just above 50 secs improves prophylaxis in high-risk cases. Furthermore, an increased dose of heparin is required than is proved during the conventional low-dose regime of 5,000 units tds. With regular control using the standardised APTT, increasing the dose to the target value does not increase post-operative haemorrhage. Further studies with larger numbers of patients are required in order to show a significant reduction in the incidence of post-operative deep vein thrombosis in hip surgery patients receiving low-dose adjusted heparin.

Adult

Dosage and control of oral anticoagulants: an international collaborative survey.

An international survey of oral anticoagulant dosage has been carried out comparing the mean dosage prescribed in hospitals in 23 countries. In addition, participants using the Quick prothrombin time test were asked to assess the adequacy of dosage of a lyophilized test plasma which was mid-therapeutic using the British Comparative Thromboplastin (BCT). The overall mean dosage proved similar for the groups of laboratories using the Quick test and human brain thromboplastin and Thrombotest although wide differences existed between individual centres. The survey indicated that these discrepancies were due partly to the adoption of different intensities of anticoagulation. In addition, local differences in patients' response to anticoagulants were apparent, e.g. North American centres prescribed a higher mean dose with a more intense therapeutic range than Europeans. Hong Kong physicians appear to prescribe a much lower dose than the rest of the world although the intensity of their treatment is comparable, whereas South African hospitals give moderate doses of warfarin despite a conservative therapeutic range. Such geographical variation in response would invalidate standardization of anticoagulant treatment based on the mean dosage approach.

Administration, Oral

An assessment of an amidolytic assay for factor VII in the laboratory control of oral anticoagulants.

A comparison has been made between the prothrombin time test using British Comparative Thromboplastin (BCT) and a chromogenic substrate assay for factor VII in the assessment of laboratory control of oral anticoagulants in short-term and long-term patients. Opportunity was also taken to compare the findings with parallel results obtained with the venous Thrombotest technique and a specific clotting assay for factor VII. There was good agreement between the amidolytic factor VII assay, using a method modified from Seligsohn et al (1978) with the Quick test using BCT and Thrombotest in 60 long-term patients. Tests in 53 patients within the first 3 weeks of starting oral anticoagulant administration gave less satisfactory agreement between the above amidolytic method and the conventional tests. In contrast, there was a good correlation between the two conventional tests in both groups and also between the clotting and amidolytic factor VII method. Although the results are an improvement on previous, less satisfactory correlations between the BCT prothrombin time method and amidolytic assays for factor II and X, the present study indicates the limitations of a specific clotting assay versus a broad spectrum extrinsic clotting test in oral anticoagulant control. While not warranting the routine use of the chromogenic assay for factor VII in place of the prothrombin time using BCT, the factor VII amidolytic assay offers a limited but dependable guide to dosage in long-term patients. The complexity of the technique in its present form militates against its adoption for routine anticoagulant control in hospital laboratories.

Acenocoumarol

Prostacyclin-like, and kallikrein activity of amniotic fluid in pre-eclampsia.

Amniotic fluid from patients with pre-eclampsia was compared with samples obtained from normotensive controls with respect to the inhibiting effect on platelet aggregation (PGI2-like activity) and activating effect on the plasma kallikrein assay and Russell's viper venom test. After 39 weeks gestation, amniotic fluid from pre-eclamptic patients showed significantly less PGI2-like activity ( p less than 0.01) and significantly lower kallikrein levels (p less than 0.01) than that from normotensive controls. The study suggests that the biosynthesis and release of PGI2-like activity and kallikrein may be impaired in pre-eclampsia. In view of the association of pre-eclampsia with intravascular clotting, the highly significant reduction of PGI2-like activity seems important and appears to warrant a clinical trial of prostacyclin administration in this disorder.

Amniotic Fluid