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Aya Sugiyama,1 Teruhisa Fujii,2,3 Yoshiko Okikawa,4 Fumie Sasaki,5 Masazumi Okajima,3,6 Hidekuni Hidaka,3,7 Koji Iwato,3,8 Kazuyoshi Sato,3,9 Akira Kokubunji,3,10 Noboru Takata,3,11 Masahiro Yamamoto,3,12 Junko Tanaka1,3 1Department of Epidemiology, Infectious Disease Control and Prevention, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan; 2Division of Transfusion Medicine, Hiroshima University Hospital, Hiroshima, Japan; 3Joint Committee for Blood Transfusion Therapy in Hiroshima Prefecture, Hiroshima, Japan; 4Department of Oncology, Hiroshima City Hiroshima Citizens Hospital, Hiroshima, Japan; 5Division of Laboratory, Hiroshima City Hiroshima Citizens Hospital, Hiroshima, Japan; 6Department of Surgery, Hiroshima City Hiroshima Citizens Hospital, Hiroshima, Japan; 7Division of Surgical Services, Fukuyama City Hospital, Hiroshima, Japan; 8Department of Hematology, Hiroshima Red Cross Hospital & Atomic-Bomb Survivors Hospital, Hiroshima, Japan; 9Division of Laboratory, Shobara Red Cross Hospital, Shobara, Japan; 10Department of Medical Science and Technology, Faculty of Health Sciences, Hiroshima International University, Hiroshima, Japan; 11Oda Internal Medical Clinic, Hiroshima, Japan; 12Hiroshima Red Cross Blood Centre, Hiroshima, JapanCorrespondence: Teruhisa FujiiDivision of Transfusion Medicine, Hiroshima University Hospital, 1-2-3, Kasumi, Minami-ward, Hiroshima, 734-8551, JapanTel +81-82-257-5581Email terchan@hiroshima-u.ac.jpPurpose: Given the chronic shortage of blood for transfusion in Japan, promotion of appropriate use of fresh frozen plasma (FFP) urgently needs to be addressed by the national blood project in Japan. Whether FFP transfusions are administered appropriately in Japan is currently unclear. In this study, we aimed to investigate the outcomes of patients who undergo FFP transfusion and the appropriateness of use of FFP.Patients and Methods: This multicentre, prospective, observational cohort study was conducted from September 2017 to April 2019 at the 15 medical institutions in Hiroshima Prefecture that are the top providers of FFP. All patients who underwent FFP transfusion during the study period were included, relevant data being extracted from the medical records. The indications for FFP transfusion were classified in accordance with the Guidelines of the Ministry of Health, Labour and Welfare of Japan. Factors associated with patient outcomes at day 28 after FFP transfusion were subjected to multivariable logistic regression analysis.Results: In total, data of 1299 patients were eligible for analysis. At least 63.8% of indications for FFP were in accordance with the guideline for FFP transfusions. The mortality rate at day 28 after FFP transfusion was 16.2%. Older age (65– 74 years: adjusted odds ratio [AOR]=4.3, ≥ 75 years: AOR=4.1), non-perioperative use (AOR=4.5), coagulopathy associated with liver damage (AOR=2.7), large volume of FFP transfused (AOR=2.5), and lack of improvement in blood coagulation following FFP transfusion were independently and significantly associated with death within 28 days after FFP transfusion.Conclusion: Our findings do not support the simple conclusion that FFP transfusions contribute to prognosis. However, given that coagulopathy in patients with end-stage liver disease is infrequently improved by FFP transfusion, “inappropriate” use of FFP should be avoided. It is important to promote appropriate use of FFP so as not to waste blood resources.Keywords: prognosis, coagulopathy, inappropriate use, compliance with guideline

BACKGROUND Granulocyte mobilization and harvesting, the two major phases of granulocyte collection, have not been standardized. STUDY DESIGN AND METHODS The data on 123 granulocyte collections were retrospectively investigated for the effect of the mobilization regimen and the harvesting technique. After a single subcutaneous dose (600 µg) of granulocyte-colony-stimulating factor (G-CSF) with (n = 68) or without (n = 40) 8 mg of orally administered dexamethasone, 108 granulocyte donors underwent granulocyte collections. Moreover, 15 peripheral blood stem cell (PBSC) donors who had received 400 µg/m2 or 10 µg/kg G-CSF for 5 days underwent granulocyte collections on the day after the last PBSC collections (PBSC-GTX donors). Granulocyte harvesting was performed by leukapheresis with (n = 108) or without (n = 15) using high-molecular-weight hydroxyethyl starch (HES). RESULTS Granulocyte donors who received mobilization with G-CSF plus dexamethasone produced significantly higher granulocyte yields than those who received G-CSF alone (7.2 × 10(10) ± 2.0 × 10(10) vs. 5.7 × 10(10) ± 1.7 × 10(10) , p = 0.006). PBSC-GTX donors produced a remarkably high granulocyte yield (9.7 × 10(10) ± 2.3 × 10(10) ). The use of HES was associated with better granulocyte collection efficiency (42 ± 7.8% vs. 10 ± 9.1%, p

In order to confirm engraftment of sibling-donor marrow cells or lekuemia relapse from the patient leukemic cells, DNA from nucleated cells in peripheral blood or bone-marrow fluid from patients who had received bone marrow transplantation (BMT) was examined using the MCT118 gene (D1S80) (a variable number of tandem repeats: VNTR) and 6 other genes and 9 restriction enzymes before and after BMT. In 5 of 10 patients (Case 1, 3, 4, 7, and 9) a difference between the DNA from donor cells and recipient cells was confirmed using the MCT118 gene, and engraftment of the donor marrow cells could be demonstrated. In Case 7, not only the engraftment of donor cells but also the leukemia relapse from recipient cells were confirmed using MCT118. Among the remaining 5 cases, Case 5, 8, and 10, the engraftment of donor cells was confirmed using the COLA2A1 gne (12q14.3). In Case 5, and 10, the engraftment could be detected using a gene in the CA2 locus (8g22). In Case 2, the presence of cells from the sibling was identified with the THRB gene (3p24) alone. In Case 6, the presence of cells from the donor could be confirmed using the DMD gene (Xp21.3-21.1) alone. In Case 8, the AK1 gene (9q34.1-34.2) could identified the difference between DNA from donor cells and recipient cells only, but the difference could also be detected using the COLA2A1 gene. In none of the cases, the INS gene (11p15.5) could identify the difference between the DNA from donor and recipient cells. Although only 10 cases were investigated, the findings suggest the use of first the MCT118 gene, then the COLA2A1 gene and the other genes when trying to determine engraftment of donor marrow cells or leukemia relapse after BMT.

Granulocyte mobilization and harvesting, the two major phases of granulocyte collection, have not been standardized.The data on 123 granulocyte collections were retrospectively investigated for the effect of the mobilization regimen and the harvesting technique. After a single subcutaneous dose (600 µg) of granulocyte-colony-stimulating factor (G-CSF) with (n = 68) or without (n = 40) 8 mg of orally administered dexamethasone, 108 granulocyte donors underwent granulocyte collections. Moreover, 15 peripheral blood stem cell (PBSC) donors who had received 400 µg/m2 or 10 µg/kg G-CSF for 5 days underwent granulocyte collections on the day after the last PBSC collections (PBSC-GTX donors). Granulocyte harvesting was performed by leukapheresis with (n = 108) or without (n = 15) using high-molecular-weight hydroxyethyl starch (HES).Granulocyte donors who received mobilization with G-CSF plus dexamethasone produced significantly higher granulocyte yields than those who received G-CSF alone (7.2 × 10(10) ± 2.0 × 10(10) vs. 5.7 × 10(10) ± 1.7 × 10(10) , p = 0.006). PBSC-GTX donors produced a remarkably high granulocyte yield (9.7 × 10(10) ± 2.3 × 10(10) ). The use of HES was associated with better granulocyte collection efficiency (42 ± 7.8% vs. 10 ± 9.1%, p 0.0001).G-CSF plus dexamethasone produces higher granulocyte yields than G-CSF alone. Granulocyte collection from PBSC donors appears to be a rational strategy, since it produces high granulocyte yields when the related patients are at a high risk for infection and reduces difficulties in finding granulocyte donors. HES should be used in apheresis procedures.

Aims/Objectives: To clarify transfusion incidence of hepatitis B virus (HBV) infected blood negative for mini pool-nucleic acid amplification testing (MP-NAT). Background: Japanese Red Cross (JRC) blood centres screen donated blood to avoid contamination with HBV. However, a low copy number of HBV may be overlooked. Methods/Materials: In Hyogo-Prefecture, JRC blood centres screened 787 695 donations for HBV from April 2005 to March 2009. Of these, 685 844 were donations from the repeat donors. To detect the donors with HBV, serological tests, MP-NAT and/or individual donation (ID)-NAT were performed. To detect the recipients with transfusion-transmitted HBV infection (TTHBI), serological analysis and/or ID-NAT were performed. Results: In this study, 265 of the 685 844 repeat donations were serologically and/or MP-NAT positive for HBV. Their repository samples from the previous donation were examined in a look-back study; 13 of the 265 repository samples proved ID-NAT positive. Twelve recipients were transfused with HBV-infected blood components derived from 10 of the 13 HBV-infected donors. Only 1 of the 12 recipients was identified as TTHBI case. Seven of the 12 recipients escaped from our follow-up study and 4 recipients were negative for HBV during the observation period. Conclusion: On the basis of the look-back study among the repeat donors in Hyogo-Prefecture, Japan, donations with HBV-infected blood negative for MP-NAT occurred with a frequency of 13 in 685 844 donations (∼1/53 000 donations). However, more than half of the recipients transfused with HBV-infected blood negative for MP-NAT could not be followed up. It is necessary to establish a more cautious follow-up system.

WE report a patient who developed severe anaphylactic shock and life-threatening ventricular fibrillation immediately after starting infusion of 5% human plasma protein fractions (Albuminar-5%; ZLB Behring, Tokyo, Japan) during general anesthesia. He was diagnosed postoperatively with ahaptoglobinemia associated with haptoglobin gene deletion. We postulated that this event may have occurred as a result of allergic reaction to a precipitating antibody to haptoglobin associated with use of plasma protein fractions.

We experienced a patient who showed severe allergic symptoms immediately after blood transfusion using a filter for removal of leukocytes. To investigate the cause of this incident, we conducted a maximization test on the antigenicity of substances extracted from filters used for leukocyte removal. The tests were performed in guinea pigs. Acetone extracts were obtained from filters made by three manufacturers (A, B and C) and sensitization and evocation were tested at 10% concentration. It was confirmed that extracts from one filter (B) induced sensitization in guinea pigs. Sensitization of the extracts was also tested at 1%, 0.1% and 0.01%, and was induced at 1% but not at 0.1% and 0.01%. Next, skin irritation and sensitization of the substances were tested using mice. 5%-ethanol extracts were prepared from filters made by 2 manufacturers (A and B) and the extracts (5% or 0.5% concentrations) were injected intradermally into the auricle and the ear swelling was observed 1, 24 and 48 hours and 7 days after injection. Significant ear swelling was induced by the extracts from B-filters. In the skin sensitization test, 5% extracts were injected subcutaneously into the back of mice for 3 continuous days for sensitization, and 0.5% extracts were injected intradermally into the auricle of mice to evoke a response, after which changes in ear swelling were observed. Biphasic ear swelling observed 1 hour (immediate response) and 24 hours (delayed response) after challenge was induced by the extracts from B-filters. In conclusion, our study showed that filters used for leukocyte removal may contain substances that cause skin irritation and sensitization and that antigenicity and irritancy tests of the filters might prevent the adverse reactions after blood transfusion.