220ml of blood from each healthy donor was collected into a triple-plastic bag containing ACD or CPD anticoagulants. Platelet concentrates (PC) with approximate volumes of 20ml were prepared by centrifugation. Each PC was divided into two portions and then stored at 22°C with or without agitation. pHs of plasma, cell volumes, aggregabilities, shape change abilities and shapes of platelets on PC were studied after storage of 0-72 hours.
The initial pH of ACD-PC was 7.2 and that of CPD-PC was 7.45. During 72 hours storage, pHs of plasma in both anticoagulants-PC without agitation, decreased to 7.0. However, pH values of plasma in agitated PC increased to 7.5-7.6. pH values of CPD-PC were always higher than ACD ones. On the other hand, differences on morphological changes of platelet, the average values of platelet MPV (mean particle volume), aggregabilities by ADP or collagen and shape change abilities were not observed in two anticoagulants-PC during the periods of experiments. Storage of platelets by agitation usually resulted in better preservation of platelets with regard to discoid shape, MPV, shape change abilities than observed with nonagitated storage.
The data suggest that gentle agitation (30cycles/min. on a platform rotator) should be necessary for better storage of PC using ACD or CPD anticoagulants.

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An 18-year-old hemophiliac with factor VIII-inhibitor was admitted to our hospital with 4 episodes of intracranial bleeding for the past 2 years. The initial episode of bleeding dated at Sep. 1, 1980 was successfully treated with a preparation of activated prothrombin complex concentrate (PCC), FEIBA, as described in Thromb. Res. 22, 177, 1981. We also discussed some problems including clinical efficacy, monitoring for hemostasis and thromboembolic complications. The conclusion led us to an emphasis that nonactivated-partial thromboplastin time (NA-PTT) and r-value on thrombelastogram (TEG) were important for monitoring.
Since then, the last 3 episodes of bleeding developed successively in a short period of 6 months, were all treated as follows: On the 1st and 2nd bleeding, a nonactivated PCC (Konyne) was used initially, but the clinical and laboratory improvement was hardly shown. So, an activated PCC (Autoplex) was introduced instead, and it gave dramatic effects on both bleeding. The stortening of NA-PTT and r-value on TEG were recorded for about 8 hours after injection, and the maximum effect was shown after 2 hours. For the 3rd episode, medical treatment started with another nonactivated PCC (Proplex), and the clinical and laboratory improvement was noted, but lesser improvement was observed with Proplex. Any thromboembolic complications were not observed through the above 3 episodes.

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To elucidate immunochemical properties of Rh-Hr antigens, serological method to detect blood type activity, influence of various solubilizers on the activity, isolation and purification of the antigens were investigated. As a very sensitive method to detect Rh-Hr antigens, agglutination inhibition test was recommended in which anti-D, -C, and -e diluted 1: 800, 1: 500 and 1: 400, papainized red cells and microtitrate plates (v type) were used.
As for solubilizing reagent, extraction with Brij 35 and Triton X-100 were effective but no activity was noted in the stroma treated with SDS. However, blood type activity of stroma treated with SDS was recovered by adding of reductants such as 2-mercaptoethanol and dithiothreitol. The activity was slightly demonstrated in the preparation from D negative red cells. The influence of the reductants on the specificity of solubilized stroma was discussed. SDS solubilized stroma was fractionated with Sephadex G-50, Sepharose 4B, activated thiol Sepharose 4B and Sepharose 6B. The final preparation with D activity was proved to correspond with Band 3 in SDS polyacrylamide gel electrophoresis (PAGE) which was not stained with PAS reagent. Passive agglutination test using red cell coated with the final preparation showed that D activity was contained in Band 3. Rh-Hr blood type activities other than D were also observed in the same fraction.
To ascertain Rh-Hr blood type activities in Band 3, deoxycholic acid (DOC), a mild anionic detergent, was used for solubilization of stroma. DOC-solubilized stroma was fractionated with gel filtrations and affinity chromatography with activated thiol Sepharose 4B, resulting in that the final preparation corresponding to Band 3 in SDS-PAGE had potent Rh-Hr blood type activities. The activities were detected without reductants and the preparation from D negative red cells had no D activity.
From these results, it is concluded that Rh-Hr blood type activities are extracted with ionic and nonionic detergents and that the activities are localized in Band 3 with no PAS reaction.

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The principles of biological standardization have been discussed, particularly as they apply to the quantitation of Factor VIII. A World Health Organization (WHO) standard for Factor VIII concentrate has existed since 1970, and virtually all manufacturers now calibrate their house standards against the WHO standard, and label their products in International Units. In 1982, as the result of an international collaborative study, a replacement standard was established for Factor VIII concentrate (the 3rd. International Standard), with an assigned potency of 3.9 IU per ampoule. By the same token that concentrates should be calibrated against concentrate standards, a stable plasma standard is the appropriate standard for measuring Factor VIII in patients' plasmas. Accordingly, in 1982, WHO also established a standard for Factor VIII-related activities in plasma.

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