In whole blood stored at a lower temperature (1±1°C) than the current preservation (4-6°C), ATP and 2, 3-DPG can be maintained at higher levels. However, the cryogenic changes in plasma Na⁺, K⁺, and Hb are accelerated markedly.
In order to inhibit those changes, an appropriate volume of glycerol and mannitol mixture (GM) was added to the whole blood preserved with CPD solution, and I found that GM was able to surpress the cryogenic changes derived from the storage at 1±1°C for 42 days. This action seems to be mainly developed by glycerol and the mechanisms are thought to be as follows; 1) GM can decrease the damage of ATPase activity induced by the low temperature, so that the active transport can be maintaind under optimal conditions. 2) GM can stabilize the red cell membrane through the hydrogen bond, because GM prohibited the increases of phospholipids and cholesterol values in plasma more than control (CPD preservation).
Although ATP is thought to be related to the posttransfusion viability, ATP like 2, 3-DPG also seems to be restored to its normal value immediately after transfusion as long as the morphologic function of red cells being maintained by utilizing ATP. Therefore, it seems that the high ATP value is not merely the important point for the posttransfusion viability.
Conclusively, the key point on the storage of the whole blood at 1±1°C seems to be how to keep the favorable conditions for ATP utilization, and as to this point, GM can keep the satisfactory condition in the whole blood storage.

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The titer of hepatitis C virus (HCV) antibodies (Ab) was assayed in 55 transfused patients with haematological disorders using enzyme immunoassay (EIA) technique, and the effects of screening test of HCV Ab in blood donors of Red Cross Blood Center on post transfusion hepatitis (PTH) was evaluated.
Seropositive conversion was found in 5 out of 55 (27.3%) transfused overall patients, and in 13 out of 38 (34.2%) patients before screening test was introduced and in 2 out of 17 (11.8%) patients after. The occurrence rare of PTH was 32.3% in patients with blood transfusion before screening test was introduced and reduced to 21.3% after.
Titration of HCV Ab in γ-globulin products were also assayed. All 6 products made from the plasma before screening test was introduced had high titration, HCV Ab was negative in 2 products made from the plasma after.
Although these data indicated the screening test of HCV Ab of blood donors was effective to reduce the occurrence of PTH, more sensitive assay systems were needed to reduce the occurrence of PTH completely.

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Sera from patients with collagen diseases followed at Nagoya City University Hospital were investigated for the detection of antibody (C100-3) to hepatitis C virus (HCV), and HCV-RNA using the method of reverse transcriptase polymerase chain reaction (RT-PCR). The overall prevalance of HCV antibody positivity was 4.8% (12/252) in the patients, and significantly higher than that (1.1-1.2%) of Japanese populations of blood donors. Relative high prevalence (6.5%) was observed in patients with Sjogren's syndrome (SjS). Seven of 12 patients with anti-HCV antibodies were detected HCV-RNA in their sera. All of these patients, except one case, had chances of the infection for HCV. 5 patients, whose sera was negative result of HCV-RNA, had no history of HCV infection. Therefore, the presence of false-positive tests for anti-HCV antibodies was 2.0% (5/245) in patients with collagen diseases. Sera from patients with SjS showed the higher false-positive tests. The false-positive tests for antibodies to HCV did not correlate with the presence of antinuclear antibodies, rheumatoid factor, and hypergamma-globulinemia.

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The significance of hepatitis C virus (HCV) antibody determination as a screening test was examined prospectively by clarifying the positive rate of HCV antibody in transfusional blood and examining the incidence of posttransfusional non-A non-B hepatitis and characters of HCV antibody positive blood after HCV antibody screening in Tokushima Prefecturte.
The following results were obteined.
1. The HCV antibody positive rate in blood after conventional screening with aspartate aminotranserase (ALT) activity was 0.9% and the rate in all donors blood were 1.35%.
2. The HCV antibody positive rate in blood of more than 36 KU ALT activity was 7.6%. The positive rate in blood of less than 35 KU ALT activity was 1.2%. The HCV antibody positive rate in higher ALT activity blood showed significantly higher values.
3. No significant difference was observed in HCV antibody positive rate between HBs antigen negative group and positive group.
4. Various test values were compared between HCV positive group and negative group after conventional screening. As a result, ALT showed significantly heighter values in the HCV antibody positive group. No significant difference was observed in alkalinephosphatase, total protein, albumin, albumin-globulin-index, blood urea nitorogen, cholesterol between both groups.
5. Posttransfusional non-A non-B hepatitis occurred in 3 (3.4%) of 87 cases that received transfusion with HCV antibody screened blood. It was significantly lower than the rate before initiation of HCV antibody screening.
6. It became clear that introduction of trnasfusion blood screening with HCV antibody is effective in the prevention of posttransfusional hepatitis.

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A new red cell preservation solution is described in which the red cells may be stored for 42 and 49 days. An in vivo and vitro study of 42 and 49 days stored red cells with saline, adenine, glucose, mannitol (SAGM) preservation solution (OPTISOL, Terumo Corporation) was conducted using 20 healthy volunteers to document in vivo efficacy and analyze the validity of the ⁵¹Cr technique. Standard in vitro parameters of OPTISOL red cell concentrates were well preserved with low levels of hemolysis and high levels of red cell ATP which is compatible with their survival after 42 and 49 days storage. Osmotic pressure at hemolysis ending point of 42 and 49 days stored red cells did not change, while osmotic pressure at hemolysis starting point increased on 49 days. The red cell viscosity increased and scanning electron-microscopic studies showed that the majority of them became echinocyte and spherocyte on 49 days. In vivo autologous post-transfusion recovery was measured by using a ⁵¹Cr-labeled red cells. After 42 days of storage, post-transfusion 24-hour recovery averaged 82.3±8.4% and 48-hour recovery was 79.1±9.1%, and after 49 days, 24-hour recovery was 75.9±3.6%, but 48-hour recovery, which averaged 69.9±5.2%, was significantly lower.
The present study reports in vivo and in vitro evaluation of a new red cell storage medium which allows high levels of post-transfusion recovery and permits without significant hemolysis. The results suggested that 42 days if a more suitable shelf life than 49 days.

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The detection and identification of platelet-specific antibodies are clinically important for diagnosing and treating refractoriness to platelets transfused or neonatal alloimmune thrombocytopenia due to platelet-specific antibodies. The serologic test of platelet-specific antibodies requires the platelet antigen panels.
Storage duration of platelet antigen panels has been limited to at most for 2 months. Although Shibata and Kim developed prolonged storage antigen panels for the MPHA (mixed passive haemagglutination) test using a U-type microplate coated with platelet soluble antigen, results are still influenced by presence of HLA Class I antigens.
The MPHA test was used to assess whether or not the removal of platelet surface HLA Class I antigens were possible by chloroquine treatment on microplate coated with platelet soluble antigen. The removal of HLA Class I antigens was possible by the 0.8M chloroquine treatment method. Platelet-specific antigens (HPA-1a, HPA-3a, HPA-4a, HPA-4b, Nak^a) were present, while HLA Class I antigens were removed on chloroquine treated platelet soluble antigens.

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Anti-HPA-2a (Miy), IgG class, was detected in a woman with platelet-transfusion refractoriness. The reaction pattern was the same as that of original anti-HPA-2a (reported by van der Weerdt) against 65 platelet panels by the MPHA (mixed passive haemagglutination) test (r=1.0). This is the first report on anti-HPA-2a in Japan. The dosage effect was observed, thus the titer of Miy serum was 1: 64 against heterozygous for HPA-2a and 1: 256 against homozygous for HPA-2a. We examined 500 Japanese voluntary blood donors using anti-HPA-2a and anti-HPA-2b. The phenotype frequencies were for HPA-2a as 98.2% and for HPA-2b as 26.6%. We suppose anti-HPA-2a antibody plays an important role in platelet transfusion refractoriness like anti-HPA-2b antibody.

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We have already demonstrated that Nak^a antigen (GPIV) is expressed on the monocytes in platelet-Nak^a- subjects. To define the molecular basis for the discrepancies of GPIV expression between monocytes and platelets in Nak^a- subjects, we studied whether there are differences in monocyte GPIV mRNA obtained from Nak^a- and Nak^a+ subjects, and compared these monocyte GPIV mRNA with Nak^a- platelet GPIV mRNA which we have recently reported.
In Northern blot analysis, monocyte GPIV mRNA from two platelet-Nak^a- subjects exhibited the same size of 2.9kb and nearly the same amounts as that from platelet-Nak^a+ subject. And restriction enzyme analysis of GPIV cDNAs which were amplified by a reverse transcription polymerase chain reaction (RT-PCR) showed predictable fragment length.
We have already shown that RT-PCR analysis of GPIV mRNA from both Nak^a- and Nak^a+ platelets and restriction enzyme analysis of these PCR products show no apparent differences between them. Thus, we conclude that GPIV mRNA from both monocytes and platelets in Nak^a- subjects has no apparent abnormalities and seems to be the same as that in Nak^a+ subjects.

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We have developed a double-bag system to extend the storage period of platelet concentrates (PC) at liquid condition. The double-bag system consists of two compartments in which the inner compartment is separated from the outer one by a semipermeable membrane. One hundred ml of PC prepared by platelet apheresis was put in the inner compartment and 500ml of the synthetic medium for platelet storage developed by Holme et al. was put in the outer compartment. To adjust the colloidal osmotic pressure, 2.5% w/v dextran was added to the platelet storage medium. The double-bag was vertically rotated at 5rpm at an angle of 45° in a 22°C incubator under normal air atmosphere.
The medium in the outer compartment was not changed during the storage period. As a control, the same volume of PC was put in polyolefin containers and stored on a horizontal agitator at 22°C.
The PC stored in the double-bag system could maintain platelet count and mean platelet volume as well as those preserved in the polyolefin containers. In contrast hypotonic shock response and aggregability of platelets were much better preserved when plateles were maintained by this new system than when stored in the polyolefin containers at the 10th day of storage, and there function retained even after 21 days.

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A new freezing system for preparing high quality source plasma was investigated. The MPIII™ portable freezing system consists of a freezer called the “Cell Unit” and a storage case. The MPIII cell unit and the MPIIIs cell unit, about half the size of the MPIII, were transferred to storage cases after “recharging” (storage at -40°C) and their ability to freeze apheresis plasma bags was tested. The MPIII and the MPIIIs freezers could freeze 8 and 4 plasma bags respectively to a temperature below -20°C. However, the MPIII could freeze 16 bags and the MPIIIs 9 bags, before the cell unit temperature rose up to -20°C. They could freeze and store plasma bags below -20°C for about one day when there were 12 bags in the MPIII or 6 bags in the MPIIIs.
The recovery of factor VIII activity (FVIII: C) of the plasma frozen in the MPIIIs was high (97%) and was similar to that of plasma frozen in a blast freezer. This high FVIII: C did not decrease even after overnight storage in the MPIIIs.
In addition, the MPIII™ portable freezing system has several other merits. No electricity is required after “recharging” the cell unit, it is safe because neither liquid nitrogen nor ethanol is required and the plasma bags are flat after freezing, which allows easy bar code reading and safe, efficient storage and transportation.
Thus it is possible for the MPIII and MPIIIs systems to freeze the collected plasma at blood collection sites and to retain high FVIII: C.

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This is a case report of a hemophilia B patient who became seropositive for HIV-1 more than 3 and a half years after switching to heat-treated prothrombin complex concentrates. This 17 year old boy has been treated in our institution since 1973, when he was a year of age. He became HIV-1 seropositive for the first time in July, 1990. Because of the delayed HIV seroconversion, we studied his serum samples stored separately, using RNA-PCR (polymerase chain reaction) method. All of his serum samples tested were positive for RNA sequences of HIV, and the earliest positive sample was obtained in August of 1985. In Japan, prothrombin complex concentrates were switched to heat-treated ones, in January, 1986. Therefore, we conclude that this is a case of hemophilia who seroconverted after a prolonged window period, showing that silent infection is also possible in hemophilia patients at risk as well as in homosexual men, although the incidence must be low (less than 1%).

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Solid phase adherence assay (SPAA) was developed for the detection of irregular IgG red cell antibodies.
The procedure of SPAA are as follows: 1) Antibody screening red cells were coated to the wells of U-bottom, protein adsorptive microplates by centrifugation. 2) Low ionic strength solution and test sera were added to the wells and incubated. 3) After washing plates, anti-human globulin reagent and IgG coated indicator red cells were added, and the plates centrifuged.
Warm type IgG antibodies considered to be clinically important were completely detected by SPAA. Moreover, antibody titers obtained with SPAA were significantly higher than with conventional LISS-antiglobulin test.
The great advantage provided by this SPAA is the easiness of reading compared with hemagglutination methods because of the use of red cell adherence as an end point.
We consider that SPAA is suitable for pre-transfusion tests since it has the following characteristics: sensitivity and specificity are good, and procedure is simple, especially on reading.

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Transfusion of leukocyte-depleted platelet concentrates prevents or delays HLA immunization in multitransfused patients. The authers devised a polyvinylchloride triple-bag system with a filter for preparation of leukocyte-depleted platelet concentrates (PC) from apheresis platelet-rich plasma (PRP). We spliced a Imugard PL filter (Terumo Corporation) into the tubing between the primary 1-liter bag and the second 1-liter bag using a sterile connection divice (SCD). Apheresis PRP was filtered through the Imugard PL in a closed system by gravity with neither priming before filtration nor rinsing after filtration.
The platelet recovery was 94.4±3.8% and leukocyte removal was 99.8±0.4%. The functions of platelet were not affected by the filtration. The filtered PRP was then centrifuged and yield leukocyte-depleted PC and plasma for freezing as fresh-frozen plasma. PC was stored in the 1-liter bag at 22°C with flatbed agitation for up 96 hours of storage. Mean platelet count did not change during the storage. pH, collagen-induced platelet aggregation response and plasma LDH activity became significantly higher than that in the ordinary PC.% HSR, MPV and morphology score were approximately the same in comparison with the ordinary PC. It may be concluded that the method is effective in preparing leukocyte-depleted PC from apheresis RPR.

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The purpose of this study is to advance the appropriate use of red cell products (whole blood+concentrated red cells) which are derived from whole blood donations to Red Cross Blood Centers in our country. We surveyed the effective utilization of red cell products in 1008 hospitals which have blood products in their deposit systems, by reviewing the number of units delivered and administered, and the number of units delivered, not administered, returned to the Blood Center and then finally discarded.
The usage rate of red cell products among delivered units in each hospital showed a remarkable variation from 100% to 5.5%. There was no correlation between usage rate and the following three factors: (1) number of beds in each hospital, (2) number of delivered red cell product units. (3) time needed for transportation of blood products from the Blood Center. 127 hospitals having blood transfusion service units had a high usage rate of red cell products and also appropriate use of deposited blood products. On the other hand, hospitals not having blood transfusion service units showed a low usage rate of red cell products.
It is therefore concluded that hospitals having blood deposit systems should establish transfusion service units in their facilities in order to achieve appropriate use of blood and blood components.

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The 1990th year's annual meeting of workshop on red cell grouping test within the jurisdiction of Central Blood Center, The Japanese Red Cross Society, has been opened.
From the 4 blood centers, 4 blood preparations, each of which was suspected as “partial D” red cells, were submitted as the test samples.
On the D antigen, judgement of operators agreed with each other for 3 out of the 4 preparations. But on remained 1 preparation (sample I), judgements of them were not agreed. On this sample I, agglutination reaction by monoclonal anti-D antibody of JRC was remarkably weak but surely observed. As the results, “Partial D” as the suspected antigenicity, was proved to be false, rather, “weak D” on some specific epitopes was suitable on this sample I, presumably.
On the otherhand, one preparation which was suspected to have antigenic variation on E antigen was found.

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