It has been reported by some studies that unstable coagulation factor activities in thawed fresh frozen plasma (FFP) decreased with time. However no report has evaluated quality change in repeatedly freeze-thawed FFP. We assessed the quality of FFP derived from 400 ml of whole blood over three freeze-thaw cycles. Regarding activities of Factor (F) V, F VIII activities were decreased, and APTT was prolonged by repeated freeze-thawing, but no significant changes in pH, sodium concentration, potassium concentration, PT, fibrinogen concentration, activities of F II, F VII, F XI, VWF: RCo were noted. PT after three freeze-thaw cycles was 11.3±0.7 sec, which meets the coagulation examination standard in "Minimum Requirements for Biological Products" (less than 20 sec). Activity of F VIII after three freeze-thaw cycles remained at 0.72±0.23 IU/ml, which satisfied the European guideline (more than 0.70 IU/ml). Thrombin generation time might be prolonged by repeated freeze-thawing, but peak height and endogenous thrombin potential were not significantly reduced by freeze-thawing repeatedly.

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We conducted a nationwide questionnaire-based survey in 2013 among surgical patients who underwent transfusion of ≥10 units of red blood cells (RBC) per day (R10) in hospitals with 300 or more beds. 1) Of the 513 hospitals that indicated they had R10 patients, 353 provided the number of R10 patients as well as the amount of RBC and fresh frozen plasma (FFP) used. The remaining 160 hospitals did not provide this information. 2) A total of 473,391 patients were transfused in the 353 hospitals. Of these, there were 19,184 R10 patients (4.1% of total patients). In R10 patients, 325,173 RBC units were transfused (17.3% of total RBC used in transfused patients), and 263,701 FFP units were transfused (31.8% of total FFP used in transfused patients). 3) For the 160 hospitals that did not respond, we estimated that the total number of R10 patients and amounts of RBC and FFP used were 27,980 patients, 472,410 units, and 381,236 units, respectively. 4) We collected 5,796 detailed case reports for R10 patients from 168 hospitals. The number of R10 patients in each department was as follows: cardiovascular surgery, 2,730 (47%); emergency, 739 (13%); digestive surgery, 634 (11%); and obstetrics/gynecological surgery, 324 (6%). In these 4 departments, 77% of the blood components were used. 5) Of the R10 patients, only 102 (1.8% of reported cases) received cryoprecipitate (CRYO), and 144 (2.5% of reported cases) received fibrinogen concentrate (FIB). The fibrinogen values in the pretransfusion period were lower than those of R10 patients that did not receive CRYO or FIB. Further, the average number of units of RBC and FFP used in patients treated with CRYO or FIB were higher than in patients without CRYO or FIB. We conclude that a further systematic study is needed to confirm the hemostatic effect and clinical indication of CRYO and FIB.

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We experienced a case in which anti-D antibody was detected in an RhD-negative recipient after RBC transfusion of RhD-negative units. A total of 8 units from 5 donors was transfused. All units were analyzed by the anti-D dissociative-adsorption test and by PCR-SSP for the RhD gene at Kanto-Koshinetsu Block Blood Center, Japanese Red Cross. In one unit, a mutation was found on the 1,227 base, which is typical in DEL. There was no obvious evidence of hemolysis or other adverse effects in the lab data and clinical course after transfusion. DEL is classified as RhD-negative based on serological testing, despite the presence of weak D antigen. Several cases about one or two transfusions of DEL to an RhD-negative recipient have been reported so far. This case suggests that DEL is classified as RhD-negative blood units and may produce anti-D antibody in an RhD-negative recipient after transfusion. Therefore, it is necessary to discuss how to prevent the transfusion of DEL to RhD-negative recipients.

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We investigated the clinical course of one case to determine whether or not the level of false positive reaction detected with column agglutination technology (CAT) reflects the activity of primary chronic cold agglutinin disease (CAD). An 80-year old woman was diagnosed with primary chronic CAD, complicated with anemia and erythema of the breast. Oral prednisolone and cyclosporine A were used for immunosuppressive therapy. During the clinical course, erythema of the breast improved transiently but then became exacerbated again. The complement titer increased transiently, and the change in the complement level was linked with the activity of erythema. False positive levels on reverse typing test detected with CAT decreased when the complement level was suppressed, whereas the cold agglutinin titer showed limited change. We detected a false positive reaction on reverse typing test by means of CAT, the temperature of which was between 25 and 28°C. Investigation of the reaction between red blood cells and antibodies in this thermal range seems to be effective and feasible for the evaluation of the capability of the antigen-antibody complex. We speculate that detection of a false positive reaction on reverse typing test by means of CAT does indeed reflect the activity of primary chronic CAD.

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