To find a suitable mode of agitation for the preparation and storage of platelet concentrates (PC), we evaluated the effects of a newly designed rotary motion flat-bed agitator on the resuspension of platelets and maintenance of platelet integrity during storage, in comparison with two conventional types of agitators, the reciprocal flat-bed and elliptical agitators. PC were stored at 20-24°C for 72 hours under agitation, and changes in pH, collagen aggregation, hypotonic shock response, morphology score and lactate dehydrogenase (LDH) discharge were measured. The percentage of PC resuspended after 16 hours of elliptical, rotary and reciprocal agitaion was 85%, 82% and 77%, respectively. The pH of PC was maintained at 7.1 to 7.5 for 72 hours with any mode of agitation, but the pH varied more widely in the reciprocal group than in the other two groups. The two types of flat-bed agitation showed better preservation of platelets than elliptical agitation as evaluated by collagen aggregation. The morphology scores of PC stored on the rotary agitator were highest among the three groups after 72 hours of storage. The greatest LDH discharge was observed in PC stored on the elliptical rotaor. These data indicate that the rotary mode is more efficient than the reciprocal mode and causes less damage to platelets due to mechanical stress than the elliptical mode. We consider the rotary mode of flat-bed agitation to be preferable for preparing and storing PC.

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Adult T-cell leukemia associated antibody (ATLA-Ab) positive persons were screened by indirect immunofluorescence (IF) testing. and their peripheral blood mononuclear cells (PBMC) collected from concentrated red blood cell (CRC) were cultured in vitro with and without phytohemagglutinin (PHA-P) after the irradiation of 2000·4000 and 8000rad. The appearance of ATL virus positive PBMC during the in vitro culture for 10 days was analyzed by IF assay using mouse monoclonal antibody ATLV-19 reactive to p19 core protein of ATLV. All of 12 samples showed ATLV positive PBMC after the culture for 10 days without irradiation. Irradiated samples decreased the appearance of ATLV positive cells, and 8000rad irradiation failed to induce ATLV positive cells in 10 out of 12 cases. This inhibition of ATLV positive cell induction is well coincident with that of in vitro cell proliferation stimulated by PHA-P.
These evidences showed the possibility that the irradiation of blood product is effective for the prevention of ATLV transmission by blood transfusion.

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White cells contaminating platelet concentrates (PC) were analyzed with respect to the number of contamined red cells. PC were prepared from 200ml whole blood anticoagulated with CPD solution and then stored at 22°C with flat-bed agitation for 3 days under the blood center condition. The white cell count per unit volume in PC was positively proportional to the red cell count per unit volume, but their relationship was not high enough (r=0.70; y=10.97x-8.65; n=118). Some PC with low red cell count contained a large number of contaminated white cells. PC containing red cells below 4×10⁴per mm³ have routinely been delivered to hospitals, in which the mean number of white cell was 2.74×10⁷per PC and platelet yields of 3.33×10¹⁰. Light microscopic studies showed that most of the contaminated white cells were recognized as lymphocytes.
The rate of alloimmunization and the incidence of febrile reaction in patients who received passive transfusion of PC were discussed in relation to the absolute number of infused white cells.

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Platelet fluorochromasia cytotoxicity test (PFCT) was employed for the detection of HLA antigens on platelets. 5×10⁴ platelets which were incorporated with carboxyfluorescein diacetate (C-FDA), were at first incubated with 1μl of antiserum for 60min at room temp, and 5μl of rabbit complement was added. After 2hrs incubation at room temp, the viability of platelets was counted by inverted fluoresent microscope.
The sensitivity of PFCT was compatible with that of lymphocyte microcytotoxity test with 30min preincubation and 1hr incubation, when HLA-A2 antisera was used. ABO blood group anti-sera were shown to be weakly cytotoxic to ABO mismatched platelets.
HLA-ABC typing of platelets from 7 type 0 donors was done by PFCT using standard HLA-ABC typing trays. Almost all HLA-A and -B antigens defined by lymphocyte cytotoxicity, were identified by PFCT, while 3 HLA-A and -B antigens failed to be typed because anti-sera of these HLA specificities showed no or weak cytotoxicity. All of HLA-Bw4 and Bw6 were well typed. However, none of HLA-C antigen could be detected with platelets.
These data indicate that the quantitative differences of HLA-A, -B and -C antigens on platelets should be taken into consideration for HLA-compatible platelet transfusion.

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Clinical and coagulation effects of RCG-5 were studied by non-blinded multicenter system with regard to single or multiple infusion in patients with hemophilia A and von Willebrand's disease.
When a single dose of 30-45 units of F. VIII: C/kg of RCG-5 was given to 10 patients with severe or moderate hemophilia A, F. VIII: C showed a rise of 1.9%/u/kg 30min after infusion. The recovery of F. VIII: C was 90.6±13.8 (mean±1SD)% and biological half-life time of F. VIII: C was 12.40hr. However, no remarkable increase of fibrinogen was seen. These data were identical to those of heattreated F. VIII concentrates now available in Japan.
When a single dose of 30-55 units of RCof/kg of RCG-5 was given to 6 patients with von Willebrand's disease (Type I, 1; IIA, 2; IIB, 1 and III, 2 cases), RCof showed a rise of 3.34%/u/kg 30min to 2hr after infusion. A recovery of RCof was 131% and biological half-life time of RCG-5 was 19.7hr. The correction of the prolonged bleeding time and the secondary rise of F. VIII: C were obtained by the single infusion of RCG-5.
Next, 341 infusions of RCG-5 were performed in 239 episodes of hemorrhages (joint, muscle, renal, cutaneous, nasal bleeding etc.) in patients with hemophilia A or von Willebrand's disease and evaluated from the hemostatic and clinicopathological points of view. The initial infusion of RCG-5 for each bleeding episode was judged to be effective in 85.4% (excellent, 56.5%+good, 28.9%) and almost all (93.7%) of episodes were treated successfully.
No significant side effects of RCG-5 were seen 24hr and 3 months after infusion with regard to clinicopathological examinations. Neither F. VIII inhibitor nor von Willebrand factor inhibitor developed. Four out of 27 patients developed allergic reactions to RCG-5 which manifested as moderate urticaria and transient local itch/pain. The frequency of the side effects of RCG-5 was 9/341 (2.6%) and slightly lower than that of cryoprecipitate.
From these results it is considered that RCG-5 is a new useful preparation to treat patients with hemophilia A as well as von Willebrand's disease.

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The patients of liver resection and postoperative hepatic failure were transfused cryoprecipitate-removed plasma (CRP) and were studied in comparison with the patients transfused fresh-frozen plasma (FFP). CRP was transfused daily to 8 patients (CRP group) from the first postoperative day, FFP to 10 patients (FFP group). Each of 3 patients of postoperative hepatic failure underwent plasma exchanges using CRP and FFP alternately as replacement fluid. Coagulation studies were carried out preoperative day and 1st, 3rd, 5th and 7th postoperative days in the patients of liver resection, and before and after plasma exchange in those of hepatic failure. Compared with FFP, CRP contains 65% of fibrinogen (Fbg) level, 25% of factor VIII clotting activity, 45% of von Willebrand factor antigen (vVF: Ag) level and 50% of fibronectin (FN) level, but other factors, including various proteins, lipids, electrolytes, etc., remain at the same levels.
In the postoperative patients there were no discrepancies in the levels of Fbg, factor VIII and FN between CRP group and FFP group. And the levels of Fbg, factor VIII and vWF: Ag (examined only in CRP group) increased postoperatively. The levels of factor VIII and vWF: Ag, which are reported to increase in various liver diseases, and the concentration of Fbg decreased immediately after plasma exchange when CRP was used as replacement fluid, but the levels of these factors were also reduced to the same degree as FFP was used. The concentration of FN increased with CRP after plasma exchange, but diminished with FFP. Although further study of the effect of CRP on FN level is needed, it is concluded that CRP can be substituted for FFP in most liver diseases.

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