With the aim of improving the undergraduate education program for students in transfusion medicine, the following materials were examined and compared with the present stage of clinical transfusion medicine and the kowledge required for its practice today: (1) 1991 survey reports from the departments of transfusion medicine or the blood transfusion center in Japanese National, public, and private universities and medical colleges, (2) the guidelines for questions appearing on the National Examination for Medical Practioners published in 1993, and (3) the 1989 guidelines from the Ministry of Health and Welfare with respect to clinical transfusion. The results showed that all three were insufficient to deal with the actual conditions required in clinical practice. Physicians, after completing their undergraduate training, may encounter opportunities to learn about other areas of medicine and the technologies involved, but usually have little opportunities to do so in the case of transfusion medicine. This means that it is very important that students acquire the necessary basic kowledge and clinical techniques of transfusion medicine as a part of their undergraduate eduation program.

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The presence of microaggregates in transfusion blood is a well known cause of adverse effects in recipient patients. Few studies, however, have examined in datail microaggregate formation in Red cell Concentrate preserved in MAP solution (RC-M·A·P). This situation contrasts with the many reports concernings macroaggregates in these products.
In this study we analyzed microaggregates in RC-M·A·P stored two or three weeks using light and scanning electron microscopic observation, immunofluoressent detection of constituents, and PCR analysis. The microaggregates consisted of disrupted platelets, granulocytes, monocytes, lymphocytes, small quantities of DNA, and, importantly, fibrin fibers. This composition suggested that, in RC-M·A·P, fibrin but not DNA played a major role in microaggregate formation, causing adhesion of blood cell debris. These findings contrast with those of previous studies of macroaggregates in RC-M·A·P, which indicated leukocyte DNA to be the adhesive core.
Further, this study indicates the importance of the improved cytocentrifugation technique we developed and used here. Without this method, collection of microaggregates would have been a laborious and time-consuming proccess.

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To evaluate the safety of leukocyte removal filters for platelet concentrates (PCs), We divided 20 units of PCs into two portions and filtered each with one of two commercially available filters, Pall PL50 (Filter P) and Sepacell PLSSA (Filter S). LDH, PF4, C3a, bradykinin (BK) and PAI-1 were measured before and after filtration. LDH and PAI-1 levels remained unchanged after filtration with both filters. PF4 showed a significant decrease (p<0.01) from 1550±553 to 513±448ng/ml with Filter P, but no change with Filter S. C3a showed a significant decrease (p<0.05) from 1069±640 to 328±198ng/ml with Filter P, but increased to 1911±1265ng/ml with Filter S. BK remained unchanged at every blood sampling point with Filter S, but showed a significant increase (p<0.01) from 20.9±13.3 to 207.2±101.9pg/ml when 5 units were filtered with Filter P. The increase in BK was storage period-dependent: Y=32.2X+261.6 (r=0.434). In summary, there are differences between the two leukocyte removal filters tested here in terms of their effects on biochemical composition of PCs.

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We used an enzyme-liked immunosorbent assay (ELISA) using polyclonal IgA purified from pooled plasma as an antigen for the screening of anti-IgA antibodies in sera from patients with nonhemolytic transfusion reactions (NHTRs). Among 154 patients referred to our laboratory from April 1994 to January 1995, two were positive for anti-IgA IgG antibodies. Specificites of the anti-IgA antibodies were examined by immunoblotting. Anti-IgA2m(1) and anti-IgA1 were detected in one case each. Sera from both patients contained normal levels of total IgA. In one case, a 43-year-old woman with chronic myelogenous leukemia showed urticaria followed by wheezing dyspnea and fever one hour after the start of platelet concentrate (PC) transfusion. In the other case, a 45-year-old woman with aplastic anemia complained of itching, urticaria, facial edema and nausea immediately after the start of PC transfusion. No antibodies against HLA, platelet-specific antigens or irregular antibodies against red blood cell antigens were detected in either patients' sera collected before transfusion or in the transfused blood in these cases. These findings suggested that these NHTRs might have been caused by subclass and allotype-specific anti-IgA antibodies. These results show that ELISA and Western blotting are useful in the detection of anti-IgA antibodies responsible for NHTRs.

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A 30-year-old woman admitted to this institution with symptoms of anemia was diagnosed as having Evans' syndrome. During the course of transfusion therapy, autoantibody specificity to anti e developed. Treatment was continued with transfusion of same blood type (CCDee cells). The patient developed hepatic dysfunction and died. Retrospectively, continuation of transfusion with the same blood type (CCDee cells) may have been inffective in this case, and possibly resulted in an increase in the number of units of blood transfused, liver dysfunction due to hemosiderosis, or other adverse effects on prognosis.

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To calculate the total “potential” frequency of preoperatively donated autologous erythrocyte (red blood cell or RBC) units, we studied 438 surgical admissions who received transfusions for surgical procedures in one year in Saitama Medical Center.
Eighty-five patients received 317 autologous RBC units, representing 3.6% (317 of 8, 823) of all RBC transfusions at our hospital in one year and 14.6% (317 of 2, 172) of all RBC transfusions for surgical procedures in the study period. Emergency operations represented 168 (38.4%) admissions, and 7 admissions were children under the age of 4. Of the remaining 263 elective surgical admissions, 85 (32.3%) cases donated autologous RBC units, 49 (18.6%) cases were anemic with preoperative hemoglobin concentrations less than 10.0g/dl, and 79 (30.3%) cases were senile and older than 68 years, second operations for malignant tumor or operations for metastaic regions, or were cases of infectious disease. The latter 128 cases were considered ineligible for preoperative autologous blood donation (PABD) and the remaining 50 (19.0%) were considered eligible for PABD. The maximum percentage eligibility for PABD would be 51.3% (135 of 263 elective surgeries), if all eligible patients had predeposited autologous blood. In conclusion, 63.0% (85 of 135) of eligible patients actually predeposited autologous blood in our hospital.

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Optimal management of neonatal alloimmune thrombocytopenia (NAIT) requires the determination of fetal platelet antigen status as early as possible. We explored the feasibility of fetal HPA-4 genotyping by amplification of DNA from amniotic fluid cells using PCR-SSP. Three fetuses in which fetal HPA-4 type from amniotic fluid cells was completely concordant with typing using the blood of these infants were studied. Two babies were shown to be homozygous HPA-4a/a, and had normal platelet counts. The third fetus was heterozygous HPA-4a/b incompatible with maternal antibody, and developed mild thrombocytopenia (116, 000/μl). These findings suggest that HPA genotype can be reliably and rapidly determined from amniotic fluid by DNA amplification.

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The objective of this survey was to study the present status of autologous blood transfusion practice from April 1994 to March 1995 in Akita Prefecture and to evaluate changes in status by comparison with our previous survey in 1993. A questionnaire was sent to 150 surgical divisions of 37 hospitals with more than 50 beds in Akita Prefecture.
A total of 132 answers (88%) were collected and evaluated. Autologous blood transfusion programs were conducted in 49 divisions (37.1%) in total, this value being increased compared with the previous result. In addition to enforcement in cardiovascular surgery and orthopedic surgery divisions, autologous blood transfusion was also widely applied in another surgical divisions such as general and neurological surgery and gynecology. No homologous blood was transfused in 84% of total cases. In general, in orthopedic and neurological surgery, and in gynecology divisions, homologous blood was used in less than 10% of cases, with predeposit autologous blood. Ninety-six divisions (73%) answered that they have instituted or are planning to institute an autologous blood transfusion program. However, 44% of general surgery divisions and 58% of neurological surgery division have no plans to introduce autologous blood transfusion due to the specific status of patients. As in the previous survey, several practical problems such as a shortage of manpower or time to participate and the necessity of cooperation with the Japan Red Cross Blood Center were also noted.

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To clarify the influence of recombinant human erythropoietin (r-HuEPO) on endogenous erythropoietin production, experiments using rats were designed as follows.
Firstly, to evaluate the effect of acute blood loss on endogenous erythropoietin production, five rats (Wistar; mean body weight 280g) were bled of 1.3% of body weight (g) by cardiac puncture under pentobarbital sodium (30mg/kg, ip) anesthesia, followed by periodic blood examinations, including serum erythropoietin (s-EPO) levels, hemoglobin and reticulocyte counts. Secondly, 12 rats were divided into two groups of six rats each, control and r-HuEPO-treated groups. Rats received no r-HuEPO or 200U/kg of r-HuEPO subcutaneously once a week after the first blood sampling. All rats were given iron sulphate. Blood (0.6% of body weight) was drawn from each rat at three weekly intervals. At the fourth donation, blood volume of 1.5% of body weight was drawn. The results were as follows: 1) serum erythropoietin significantly increased at 3h after phlebotomy. s-EPO level peaked at 24h after phlebotomy, then returned to normal within 7 days; 2) mean hemoglobin level in the r-HuEPO-treated group decreased gradually during the 3 weeks of blood donation; 3) after the fourth donation, the peak in s-EPO level in control group occurred on day one, whereas in half of the rats in the r-HuEPO-treated group it occurred on day 7. On the basis of our data, however, no significant difference was found in recovery from phlebotomy-induced anemia between the two groups. In conclusion, r-HuEPO seems useful for the speed correction of anemia associated with successive phlebotomy. However, our data suggest that the administration of r-HuEPO suppres endogenous erythropoietin production after acute massive bleeding. Further study remains to be done to ascertain the clinical implications of these findings.

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