External quality assessment scheme for blood coagulation.

The aim of setting an External Quality Assessment Scheme (EQAS) for Blood Coagulation is to achieve a harmonization among the participating laboratories on blood coagulation testing. In Indonesia EQAS for blood coagulation is organized by the Department of Clinical Pathology, Faculty of Medicine, University of Indonesia which cooperates with the Indonesian Society of Clinical Pathologists, Jakarta Chapter. Currently coagulation tests in Indonesia are only performed in a limited number of laboratories especially in the hospital. Therefore only 65 laboratories participated in the first trial of EQAS. The control material for EQAS was provided by Organon Teknika and parameters involved in the first trial were PT, INR, APTT, and fibrinogen. Currently there are 7 brands of reagents for coagulation tests available in the market, i.e.: Behring, Biomerieux, Biopool, Human, Nycomed, Organon, and Ortho. In the evaluation, the results of each participant were compared to the median of participants who used the same reagent. If the number of participants using a given reagent was less than 10, then the result of each participant was compared to the median of overall participants. The result of a given parameter was classified as within consensus if it fell into the range of median +/- 15%. In the overall evaluation, the percentage of participants which was classified as within consensus for PT, INR, APTT, and fibrinogen were 68%, 64%, 63%, and 67%, respectively, but the CV for PT, INR, APTT, and fibrinogen were 19.84%, 17.89%, 20.21%, and 23.96% respectively. In the evaluation of participating laboratories using Behring's reagent, the percentages of participants classified as within consensus for PT, INR, APTT, and fibrinogen were 72%, 82%, 54%, and 74%, respectively. For Organon's product users, the percentages of those parameters were 84.6%-90%, 48%-75%, 86%, and 67%, respectively. It is concluded that around two-thirds of overall participating laboratories achieved harmonization in the results of coagulation test, but the variation of all parameters is still wide.

Identification of C5+ extraband of butyrylcholinesterase and two protein bands cathodic to it.

Electrophoresis of human plasma yields 4 butyrylcholinesterase (BChE) protein bands, i.e. C1, C2, C3, C4 and in some individuals also an extraband C5+. In addition to that other protein bands called "S" bands are also invariably detected. In order to know whether the C5+ and the "S" bands are related to the BChE protein, we have carried out immunological and peptide mapping studies on these proteins. The immunology approach was done by raising polyclonal antibodies against each protein bands (S1, S2, C4 and C5+) and reacted to the plasma protein bands transferred on nitrocellulose papers. Individual raised antibodies recognized all protein bands studied including the C4, an isozyme of BChE, indicating that the protein bands contain similar epitopes. Several protein bands cathodic to S1 also reacted with the antibodies, suggesting that they are probably fractions of the BChE protein, as well. When individual protein bands were digested with S. aureus V8 toxin and α-chymotrypsin, they revealed a striking similarity in peptide pattern among each other. These studies indicate that the S1, S2 and C5+ protein bands belong to the BChE protein.

Country report: the Indonesian NEQAS on hematology. National External Quality Assessment Scheme.

Around six hundred clinical laboratories in all the province of Indonesia participated the Indonesian National External Quality Assessment Scheme (NEQAS) on Hematology (Program Nasional Pemantapan Kualitas Laboratorium Klinik bidang Hematologi). Automated analyzer gave better results compared to the manual method. For hemoglobin, the CV of automated analyzer and manual method were 2.8% and 9.1%, respectively. The CV of automated analyzer and manual method for leukocyte count were 8.3% and 32.3%; for erythrocyte count were 9.7% and 80.8%; and for thrombocyte count were 10.3% and 45.9%. We observe no significant improvement of the overall performance from 1986 to 1998. Quality control material for NEQAS on hematology is still a problem. The artificial particles seem not behave exactly like the human cells (leukocytes, thrombocytes).

PF3 activity in normal subjects and beta-thalassemia trait.

Platelet factor 3 (PF3) is a platelet membrane component that plays an important role in the activation of the coagulation mechanism. Whenever platelet activation occurred, PF3 is released and participates in thrombin formation. Erythrocyte membrane fraction has also some PF3 like activity, and in abnormal erythrocyte membrane disorders, eg thalassemia, some of the membrane fraction accelerates platelet activation by increasing the PF3 activity. Formerly it was difficult to measure the PF3 activity in plasma. Recently a sensitive chromogenic test to determine the PF3 activity, which could detect the changes in PF3 activity with time, was introduced. This study was done to observe the effect of abnormal erythrocyte on platelet activation. The results obtained using the chromogenic method are the following: whole blood taken from normal subjects showed OD 0.11 +/- 0.06 at 0 minutes after blood collection and then increased significantly (p < 0.01) to 0.21 +/- 0.10 after 90 minutes, while the platelet count did not differ significantly (p > 0.05). Those results showed that there were some platelet activation after 90 minutes as seen by the increased PF3 activity, with no significant change in platelet counts. In beta-thalassemic trait subjects the PF3 activity in whole blood at 0 minutes did not differ significantly compared to the normal subjects, but after 90 minutes it was significantly higher (p < 0.01), OD 0.52 +/- 0.35. However the PF3 in platelet rich plasma at 90 minutes did not increase. The platelet count after 90 minutes was significantly decreased (p < 0.01) This result suggest that the increase in PF3 activity was caused by the role of the abnormal erythrocytes.

The effect of Norplant on some hemostatic parameters in Indonesian women.

Many reports have indicated that oral contraceptives can increase the incidence of thromboembolic disorders. Norplant, an implant contraceptive containing levonorgestrel, has been developed recently. The aim of this study is to observe the effect of Norplant on some hemostatic parameters. The subjects in this study were divided into 5 groups. Group 1 (control) consisted of 25 female blood donors. Group 2 (N = 25), group 3 (n = 25), group 4 (n = 17) and group 5 (N = 20) consisted of subjects who had been using Norplant for 2, 3, 4, and 5 years, respectively. Prothrombin time, activated partial thromboplastin time, fibrinogen level, assay of F VII and X, antithrombin III activity, plasminogen activity, alpha 2-plasmin inhibitor activity and platelet aggregation test were done in all subjects. Our results showed that there was a significant difference (p < 0.05) on platelet aggregation induced by 10 microM of ADP between the control group and Norplant users for more than 2 years, while the other parameters did not differ significantly. It is concluded that 5 years users of Norplant did not alter blood coagulability, but increased platelet response to 10 microM of ADP.