The prevalence of antibodies to hepatitis C virus (HCV) was investigated using HCV Ab ELASA kit (Ortho diagnostics) on a total of 167 Japanese patients with hemophilia and its related disorders. Seropositive reaction was found in 82 out of 126 (65%) patients with hemophilia A, 10 out of 17 (59%) patients with hemophilia B, 8 out of 17 (47%) patients with von Willbrand disease, and one out of 7 (14%) patients with congenital platelet function disorders. The patients having positive HCV Ab reaction received clotting factor concentrates on a total amount of 37, 570 units annually, whereas those with the negative 23, 689 units. The higher percentage of seropositive reaction was observed in the patients aged from 10 to 50 years old. The five hemophilias showed seronegative conversion on their clinical course over 5 years, and one had transient seropositive conversion.

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Transmission of blood-borne viruses, particularly non-A non-B hepatitis virus and human immunodeficiency virus, has been a major problem in hemophiliacs. An enzyme immunoassay (Orth-HCV ELISA) for antibodies against the hepatitis C virus (HCV) was used to test the serum samples from 210 patients with hemophilia. In addition to HCV antibody test, we tested for HBsag, HBsab, HBcab, HIV, HTLV-1 and CMV antibodies.
One hundred ninty-seven received non-heat-treated concentrates and 154 (78.2%) were seropositive; of the 13 received only heat-treated concentrates, 4 (30.8%) were seropositive. Over all, one hundred fifty eight (75.2%) of 210 patients tested for anti HCV antibody had an antibody to HCV; The prevalence of anti-HCV was well correlated to that of anti-HBV, indicating that these two viruses could be infected in the similar manner. In contrast, the prevalence of anti-HCV was not higher for patients who were HIV-positive than for patients who were HIV-negative. There was also no relation to the type and severity of hemophilia. The patients who had a high titer of antibody to HCV showed elevated level of serum alanine aminotransferase (ALT), even though 30% of hemophiliacs with persistently normal ALT level had anti-HCV antibody. Six of 210 patients with persistently elevated ALT showed antibody against neither HAV, HBV nor HCV.

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Twenty-one hemophilia patients followed at St. Marianna University Hospital were investigated for the detection of antibody against hepatitis C virus (HCV). All patients had been received at least three years. We examined HCV antibody serially for those patients. The relation between titer of HCV antibody and the levels of alanine aminotransferase (ALT) was studied.
The overall prevalence of HCV antibody positivity was 85.7% (18/21 patients). Four patients went from seropositive to seronegative, however, fourteen patients were repeatedly seropositive. In these four patients, three of them showed normalized ALT levels accompanied with decreased levels of HCV antibody.
Furthermore, six repeatedly seropositive patients and three seroconverted (from seropositive to seronegative) patients were investigated to detect HCV genome by reverse transcription-polymerase chain reaction (RT-PCR) technique using NS-5 region primers diverted from the HCV nucleotide sequence (Houghton et al. European patent 0318216. 1989).
In five of the former six patients HCV genome were detected, but in the latter three patients HCV genome could not be detected.

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Platelet concentrate (PC) was prepared in triple-unit bags which has a 30ml satellite bag made of polyolefin (PO), and stored in the PO bag for 120 hours at room temperature. Plasma pH of PC stored in the PO-bag was higher than that stored in a PVC bag, although the decreases of plasma pH in both bags were slight. Platelet count, platelet volume and β-TG leakage were about equal to the control, but %HSR was apparently higher in the PO-bag. Aggregability was also higher in the PO-bag. Furthermore, in vivo recovery and survival of platelets stored in a PO-bag exceeded the results of a PVC bag. These results suggest that platelet functions are maintained well in PO-bag when pH decrease dose not occur.

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Recent studies have shown that the transfusion of leukocyte-depleted platelet concentrates (PCs) could reduce HLA alloimmunization and platelet refractoriness. The authors developed a new porous polyurethane filter for the depletion of leukocytes from PCs and evaluated its efficiency in removing leukocytes. The new filter, the Imugard PL, is designed to be used without priming at the patient's bedside. The leukocyte removal rate, platelet recovery and platelet functions were studied. The leukocyte removal rate was higher than 99.9% when a pool of 10 units of PCs stored for 1-3 days or apheresis PCs similar in number to 15 units of PCs stored for 1 day was filtered.
The platelet recovery was higher than 92.5% for the apheresis PCs and 96% for the pools of 10 PCs. The morphology and functions of platelets were not affected by the filtration except for a slight increase of the response to aggregation and a decrease of mean platelet volume after filtration. The Imugard PL is effective in removing leukocytes without losing platelets and should help to improve platelet transfusion therapy.

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We described seven thrombocytopenic patients.
The first case was a neonatal alloimmune thrombocytopenia with a platelet-specific alloantibody, anti-HPA-4b. The baby was female. This female baby's clinical conditions were always normal (no bleeding signs, no purple spot on her skin) thought her platelet count was 2.2×10⁴/μl. On the 13th day of her birth, her platelet count rose to 11.9×10⁴/μl.
The second case was a 32-year-old woman who had developed thrombocytopenia after delivery of her 1st child (7 years ago). We found anti-HPA-2b in her serum. Her husband was typed as HPA-2 (b+) and she had a history of blood transfusion. Therefore, we could not identify the cause of this platelet antibody production.
The other five thrombocytopenic patients were refractory to the transfusions of random and HLA-matched platelets. The specificity of all of these antibodies was anti-HPA-4b.
We carried out screening tests for platelet antibodies on 15, 522 female Osaka donors. Thirty-one out of the 15, 522 donors had platelet-specific antibodies. The specificities of the antibodies were 30 anti-HPA-4b and one anti-Nak^a.
In the present paper, we suggested a strong association between the presence of the HLA antigen DR2 (A24-Bw52-Cw—-DR2) and HPA-4b immunization in HPA-4b negative individials.

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We detected anti-platelet alloantibodies in the sera from blood donors by MPHA (mixed passive haemagglutination). Anti-Yuk^a antibodies were detected with high frequency (0.12%) and 15 of 21 (71.4%) donors who have anti-Yuk^a antibodies were HLA A24-Bw52-DR2 positive.
The phenotype frequencies of Br^a (HPA-5b) and Br^b (HPA-5a) were estimated to be 8.5% and 99.8% in the Japanese population. The gene frequencies for Br^a (HPA-5b) and Br^b (HPA-5a) were estimated to be 0.043 and 0.959 respectively.

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A gelatin particle agglutination (PA) assay for detecting serum antibody to cytomegalovirus (CMV-Ab) was established. CMV antigen extracted from CMV infected human embryonic lung fibroblast (HEL) cells was coated on gelatin particles. These particles were mixed with diluted test serum in 96 well round bottom microtiterplates. After 2h incubation at room temperature, the positivity of test serum was judged by the sedimented pattern of agglutinated particles in the wells. The sensitivity and the positivity of this PA were compared with an enzyme immunoassay (EIA) and a latex agglutination (LA) assay. The results of the PA assay were well coincident with those of the EIA. On the other hand, the LA assay was less sensitive than the PA assay and the EIA. These results indicated that the PA assay had good specificity and sensitivity for the detection of CMV-Ab in sera. Furthermore, simplicity of the PA assay was considered to be advantageous for the mass screening of blood donors. A total of 2553 blood donors were tested using PA assay at Aichi Blood Center. The CMV-Ab positive rate was 85%, and the positive rate increased with age (18-20 y. o. 61%; 61-64 y. o. 100%).

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Intraoperative blood loss during marrow harvesting were approximately more than 1000ml for allogenic and autologous bone marrow transplantation. The leap flog technique was used in 8 cases with saving around 1100ml of blood before operation. All cases were not required homologous blood transfusion. We concluded that the blood saving technique for autologous blood transfusion is effective and safe, since homologous transfusion is not utilized, and the possibility of alloimmunization and the risk of infection are negligible.

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In autotransfusion, the limitation of blood volume preserved is the major problem in liquid storage of blood donated for elective surgery. Using the Sterile Connection Device (SCD, Du-Pont, U. S. A.) for 64 patients, it was possible to collect on an average 1179ml of autologous blood by an average of 5 times of venipunctures. Experimentally no bacterial contamination was detected in the media of the connected tubes.
By this new method avoiding multiple venipunctures for collecting and infusing autologous blood, enough blood volume can be collected.
We conclude that it is very useful in donating autologous blood especially for pediatric patients, small adults and patients with narrow veins.

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Since April 1986 we have made a marked effort to promote the proper use of blood and blood products based on the guidelines enacted by the Ministry of Health and Welfare in 1996. Our effort was conducted by means of discussion and technical cooporation with surgeons and physicians in Kyoto hospitals. In order to evaluate the results of our effort, we reviewed the number of units delivered to hospitals and transfused, and the number of units delivered, not transfused and returned to the Blood Center and finally discarded.
The ratio of transfused concentrated red cells derived from 400ml whole blood donation (400-CRC) in total of. 400-CRC and 200-CRC increased from 28.2% in 1986 to 53.0% in 1989. The use of whole blood decreased slightly, and conversely the use of CRC increased during the same period. In particular, the use of fresh frozen plasma (FFP) decreased remarkably and the FFP/CRC ratio changed from 1.7 in 1986, to 1.3 in the first half of 1990. The use of blood and blood products was revealed to be more appropriate in several hospitals having an established blood transfusion service unit.
We think promotion of the proper use of blood and blood products should be emphasized continuously especially in the surgical field under the powerful guidance of the Ministry of Health and Welfare.

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A Japanese girl with blood phenotype O who is complicated with a rare deficiency of selective anti-A agglutinin is described. (A) Tests for patient red blood cell (RBC): Commercial anti-A or anti-B serum did not agglutinate the patient RBCs. Neither anti-A nor anti-B agglutinin activity was recovered from patient RBCs by absorption-elution experiment according to the method of Landsteiner & Miller. Flowcytometric analysis revealed that patient RBC is totally lacked of immunoreactive material against both anti-A and anti-B antibodies. (B) Patient serum: Patient serum preferentially agglutinated RBCs from type B individuals, but not from type A individuals. Both the activities of A-and B-transferase in the patient serum were undetectable. In patient saliva, excretion of H-substance was indicated by inhibition test using anti-H lectin (Ulex-lectin). Furthermore, neither the presence of irregular antibodies nor quantitative decrease of immunoglobulins G, A, and M was observed in patient serum. Studies of her family members confirmed that the propositus was an only patient having blood phenotype O which lacked anti-A agglutinin activity despite of normal expression of anti-B agglutinin.

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