A long-term, multi-center clinical study (Stage III) was performed in 20 patients with moderate or severe hemophilia A to evaluate the safety and efficacy of a recombinant human factor VIII preparation, BAY w 6240 (rFVIII). Five had participated in a pilot study (Stage I/II) and then entered into this study, and 15 subjects were newly enrolled. One subject discontinued 1 month after treatment and efficacy analysis was excluded in this subject. Treatment period of rFVIII ranged from 24 weeks to 112 weeks.
The mean T1/2 value for rFVIII in the newly enrolled patients was 18.0 hours (n=14), comparable to that for plasma-derived FVIII concentrates. The mean in vivo recovery rate with rFVIII was 72.3% (n=14), comparable to that with plasma-derived FVIII concentrates.
Five hundred and sixty-six bleeding episodes were assessed during this study. Hemostatic effect was confirmed with an efficacy rate of 98.8%, which was similar to our pilot study results. Five surgical operations (2 for transurethral resections of bladder tumor and 3 for teeth extractions) were also managed satisfactorily by rFVIII treatment. Only two adverse reactions (0.2%), mild nausea and local vascular pain attributed to rFVIII, were observed in a total of 1247 infusions. Neither FVIII-inhibitor nor antibodies to foreign proteins were detected. No significant changes attributed to rFVIII were seen in vital signs or laboratory findings.
These results led to the conclusion that rFVIII is efficacious and safe for long-term replacement therapy for hemophilia A.

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It has been indicated the highly purified factor VIII concentrates may be activated through the purification process. For the purpose of examining the activation in the factor VIII concentrates, the relation of factor VIII activated by thrombin to the potency in plasma-derived factor VIII (Hemofi1M) and recombinant factor VIII (BL-160, Baxter). The potency was measured by one stage clotting assay (VIIIC) and amidolytic assay (VIIIam), and then VIIIam/VIIIC ratio was calculated. At same time, factor VIII proteins were monitored by immunoblotting assay. Before thrombin activation, the ratio was 1.0 approximately, but it became 0.2-0.1 after the activation. The ratio of both products (3 lots) was within a range of normal plasma (n=30). L chain cleavage of BL-160 by thrombin was slower than Hemofi1M. But L chain was cleaved more rapidly, when vWF was added to BL-160.
These results suggest that the ratio may be useful as a parameter to estimate the degree of activation in factor VIII concentrates. In addition it is assumed that vWF contributes to the cleavage of L chain, and that vWF, when added to BL-160, gives higher potency by VIIIC.

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The number of residual leukocytes in blood filtered through a leukocyte removal filter was determined by flow cytometric technique after concentrating specimens (a sample concentration method). In this method, propidium iodide (PI) was used to stain the nuclei of leukocytes remaining in the blood. To determine the number of leukocytes in red cell concentrates (RCC), seven times as much PI solution (3500μl) as volume of specimen (500μl) was added, and the suspension was centrifuged. After removal of the supernatant, it was resuspended in 400μl of PI solution. With platelet concentrates (PC), 1.5 times as much PI solution (3000μl) as the volume of specimens (2000μl) was added, and the specimen was centrifuged, the supernatant removed, and the sample resuspended in 400μl of PI solution. The regression line obtained after concentration was: y=0.770x-27 (r=0.986, n=21) for RCC and y=0.755x-20 (r=0.970, n=38) for PC. The mean leukocyte removal rate revealed 99.97±0.02% by filtration of 100ml RCC using a Pall RC 100 filter, and 99.90±0.07% by filtration of 20 units pooled PC using a Pall PL100 filter. The concentration method used in the present study caused no silting in the flow cytometer which can result from insoluble aggregates generated by concentration. It increased the sensitivity for determination of extremely low numbers of leukocytes. This method will also provide an effective means of increasing the sensitivity of other types of flow cytometries.

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Knowing the population of residual leukocytes in filtered blood products is as important as knowing the number of leukocytes to elucidate the mechanisms of the side effects caused by transfused leukocytes and to find ways to prevent them completely. We concentrated residual leukocytes in the filtered red cell concentrates (RCC) and platelet concentrates (PC) by centrifugation with Ficoll-Paque and Percoll respectively, and analyzed the differential population of leukocytes, as well as subpopulations and subsets of lymphocytes using a flowcytometer. Among leukobytes collected from the filtered PC, monocytes were found to be most effectively removed by all the filters tested in this study; the Pa11 PL100, Sepacell PL5N, Imugard PL and Imugard IG-400Y. When using filters made for the RCC, Imugard IG-400Y, Pall RC100, Sepacell R-500N, Imugard RC, monocytes are also most effectively removed. Granulocytes tended to come out of filters made of non-woven polyseter fibers (Sepacell R-500N and Pall RC100) if the RCC was fresh and filtered above 4°C. B cells were removed more effectively than T cells by filters made for RCC. On the other hand the specificity to the B cells of filters made for PC was much less than that observed using the filters made for RCC. CD8⁺ cells were more specifically removed by the filters made for PC. Among the RCC filters, the Imugard RC filter made of microporous polyvinyl alcohol could remove CD8⁺ cells most effectively. The filters made for RCC removed NK cells more effectively than T cells, but the filters made for PC did not show specificity to NK cells.
This study should help to clarify the mechanisms of alloimmunization and viral infection transmitted by specific leukocytes and also to elucidate the mechanism of leukocyte-depletion by filters, which are not perfectly understand at present.

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To see whether the introduction of screening for hepatitis C virus (HCV) would be worthwhile, the incidence of post-transfusion non-A, non-B hepatitis (PT-NANBH) was assessed among patients receiving blood during operation at Fukushima Medical College Hospital.
230 patients before screening and 108 after screening, who received blood from an average of 6.7 donors and 5.3 donors, respectively, were followed up and evaluated. Remarkable reduction of the incidence of confirmed cases of PT-NANBH was observed. 20 (8.7%) of the patients before screening showed biochemical evidence of PT-NANBH, whereas only 3 (2.8%) cases after screening did. In contrast to confirmed cases, there was a little reduction rate of suspected PT-NANBH by the screening, 16.1% before and 12.0% after screening, respectively.
These results suggest that mass-screening for anti-HCV is effective but still not perfect in reducing the PT-NANBH.
Moreover, irradiating the blood positive for anti-HCV had no protective effect in the incidence of PT-NANBH in recipients.

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We examined the effect of donor blood screening of HCV antibody on the reduction of the incidence of posttransfusion hepatitis (PTH).
Before this screening period, from September 1 to November 13, 1989, 263 cases in Hokkaido area were given with the mean dose of 11.0 units and followed for more than 3 months. Fourteen cases were diagnosed as posttransfusion hepatitis (5.3%), i. e. 9 cases definite, and 5 cases probable. Eight patients were diagnosed as hepatitis C, since either sera from patients themselves or donor blood samples turned out to be positive for aniti-HCV retrospectively.
After the screening period, from November 14, 1989 to July 31, 1990, 555 cases were given with the mean dose of 14.2 units and followed for more than 3 months. Twelve cases were diagnosed as PTH (2.2%), i. e. 5 cases definite, and 7 cases probable. In 5 cases patient's anti-HCV converted to positive and in one of those cases two of the implicated donors seroconverted after the blood donation.
In conclusion, it was revealed that incidence of PTH reduced from 5.3% (14/263) to 2.2% (12/555) significantly by HCV antibody screening of donor blood.

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Non-A, non-B post transfusion hepatitis (NANB-PTH) occurred in 10 out of 122 patients (8.2%) with orthopedic surgery between June 1989 and June 1990. The incidence of NANB-PTH did not decreased significantly after starting laboratory screening tests for antibody to hepatitis C virus, Chiron C100, in Japanese Red Cross Blood Centers (8.3% before vs 8.0% after). Mean number of blood donors was 3.9. There was no significant increase in the incidence of NANB-PTH between the groups transfused from less than 5 donors and from more than 6 donors, nor between the groups transfused from donors with ALT level below 20KU and over 21 to 35KU.
There were one donor with both anti-C100 and anti-GOR antibodies and two with anti-GOR antibody in 501 donors of blood transfused into 122 patients. Anti-C100 antibody developed in one of the patients with NANB-PTH, who received blood with both anti-C100 and anti-GOR antibodies. Another PTH patient developed anti-GOR antibody, who received only blood with neither anti-C100 nor anti-GOR antibody. Two patients transfused blood with anti-GOR antibody showed no evidence of PTH.
The current screening method of blood donors for HCV associated antibodies should be improved more in sensitivity and specificity.

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During a period from August, 1988 to March, 1991, 273 transfused patients were cheked consecutively for anti-HCV antibody, anti-HBV antibody, HB antigen, and surrogate markers for hepatitis before blood transfusion and at 2 weeks interval until 3 months or more thereafter. Hepatitis was diagnosed according to the criteria by Japan Hepatitis Research Committee made in February, 1985. According to the quality of transfused blood and blood components these patients were divided into 3 groups: Group I comprized 60 recipients of blood with and without a past history of blood transfusion, produced following the old safety standards of Japan Red Cross Blood Programme, group II 111 ones of blood without such a history and group III 102 ones of blood with no such a history and no anti-HCV antibody. Incidences of NANBH of group I, II, and III were 10.0%, 7.2%, and 4.9%, respectively. Statistical analysis made clear that the difference of incidences between group I and group III is significant (p<0.05). Our results also show clear association (11/19; 58%) between anti-HCV seroconversion and NANBH.
In conclusion, our preliminary data suggest that deferral of anti-HCV seropositive blood with a past history of blood transfusion apparently decrease the incidence of NANBH. However, the preventive effect of anti-c100-3 antibody screening as available now is not sufficient, aproximately 50%. Early development of more effective tests such as anti-HCV 2nd generation antibody test is expected urgently in our blood centers.

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In the United States, the FDA licenses Red Blood Cells Frozen (RBCF) up to 10 years and the AABB (AMERICAN ASSOCIATION OF BLOOD BANKS)'s Standards allow frozen storage of blood for routine transfusion up to 10 years.
But in Japan, the storage period of RBCF for routine transfusion allowed is 5 years and that of the thawed, deglycerolized red blood cells (DRBC) is only 12 hours at 4-6°.
The quality of RBCF stored for 5 years or 10 years was investigated to extend the storage period of RBCF and DRBC.
ATP levels and morphology scores of DRBC stored for 10 years were high as those of DRBC stored for 5 years but 2, 3-DPG levels were slightly low.
The hemolysis of DRBC immediately after deglycerolization was low, but increased rapidly with the storage. The result obtained indicated that the quality of the DRBC was maintained in good conditions. We would like to propose to extend the storage period of the RBCF up to 10 years.

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Almost four years have past since autologous blood donation program was introduced into our blood transfusion service, and an increasing number of patients scheduled for elective surgery has donated autologous blood. We experienced a total of 395 subjects as of May 31, 1991. In 1990, of the 1158 patients who underwent surgery in need of transfusion, 137 (12%) were autologous blood donors; 103 of these were operated on without homologous blood transfusion. Of the 480 units collected over a year, 86 (18%) were not used.
As more experience has accumulated, it becomes clear that candidates with the age of older than 64 years or younger than 16 years were allowed to participate in this program considering their general conditions.
Our data also indicate that the so-called “Hb=10g/dl, Hct=30% rule” for blood transfusion has become nothing but a standard, and autologous blood transfusion has also appeared to be beneficial from the viewpoint of cost-benefit ratio.

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We describe a rapid method for detection of antibodies to T-lymphotropic virus type I (HTLV-I) by using synthetic peptide. An ELISA using synthetic peptide antigen (in this case residues 100 to 130 of the HTLV-I gag p19 protein) permitted the detection of antibodies in 100 sera from blood donors within 15 minutes.
The results obtained with a rapid ELISA were identical to those in standard ELISA, requiring at least 2 hours to complete. A rapid ELISA is potentially suitable for simple, rapid, qualitative screening for antibodies to HTLV-I.

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A-43-year-old woman presented with thrombocytopenia and a positive direct antiglobulin test, and was found to have an antibody with anti-Jk^a spesificity in her serum and elute. Her red blood cells treated with chloroquine were typed as Jk(a+b+) and absorbed the antibody. And she had never had a blood transfusion, not taken any drugs. She was diagnosed as Sjögren syndrome and had no clinical or hematologic evidence of hemolytic anemia. The titration of autoantibody was 8 in her serum and 4 in her elute and was non reactive after treatment with 2-mercaptoethanol. Class and subclass of the antibody was determind as IgG₁.
Through the clinical course in this case, thrombocytopenia was improved without any treatments, titration of antibody was almost no change and any signs of hemolysis were not found. Autoanti-Jk^a antibody was not common and we were aware of only 13 previously reported examples.

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