Some artificial mediums for storage of platelets were investigated in order to improve the quality of platelet concentrate (PC). Platelets were stored at 22°C for five days in an artificial medium with or without histidine (medium C and B), which is based on Holme's method but the use of CO2 incubator, and the in vitro quality and function were compared with those of platelets stored in plasma as a control. The pH was maintained in the range of between 7.2 and 7.6 after the storage in any medium. The collagen-induced aggregability of platelets was significantly higher in medium B than in any other medium. In addition, the abilities of shape change were better in both artificial mediums than in plasma. No significant differences were observed in the number of platelets and %HSR between the three mediums. The results show that good quality of platelets is maintained in artificial medium B, which may enable us to extend shelf life of PC by further improvement.
The efficacy of a new leukocyte removal filter, Imugard E, made of polyvinyl-alcohol was compared with the Sepacell R-500N, a non-woven polyester filter. Red cell concentrates (RCC) was prepared from 400ml of whole blood, then filtered within 8 hours after collection, or on 3rd or 10th day of storage at 4°C. The effect of addition of saline to RCC prior to filtration was tested for Imugard E. After the filtration, the saline was added to the leukocyte-depleted RCC through the filter to make the final volume of the RCC to 400ml. Saline was primed for Sepacell R-500N, but not for Imugard E. Both Imugard E and Sepacell R-500N could remove more than 99.1% of leukocytes in most situations. The differences were not statistically significant. When fresh RCC was filtered with Sepacell R-500N, the leukocyte removal rate decreased to 95% because of the leakage of granulocytes, but this could be prevented by cooling the RCC at 4°C for 3 hours before filtration. The average number of residual leukocytes in the filtered RCC stored 3 to 10 days by either filter was less than 1.85×107cells/bag. The recovery of red cells in Imugard E treated RCC (saline-added) was about 90% before rinse, and it increased to 98% after rinse. These numbers were better than those in the RCC treated by Sepacell R-500N. On the other hand, the platelet removal rate was higher in the RCC filtered with Sepacell R-500N than that filtered with Imugard E. Thus, the new leukocyte removal filter Imugard E seems to have the capacity to prepare leukocyte-depleted RCC equivalent to those filtered by the Sepacell R-500N.
We investigated whether anti-human cytomegalovirus (HCMV) antibody in patients with recent episodes of HCMV infection binds to specific target antigen (s) of HCMV. A total of 330 human sera with different IgG and IgM anti-HCMV antibodies were studied for their reactivity with HCMV structural polypeptides which were separated by SDS-PAGE and electrotransferred to nitrocellulose paper. IgG anti-HCMV antibodies reacted with at least 13 structural polypeptides with molecular weights of 150, 145, 120, 86, 78, 64, 60, 50, 43, 42, 39, 32, and 30 kD in the patient's sera and that of healthy donors. The band detected by immunoblotting intensified and the number of target antigens detected by anti-HCMV antibodies increased after HCMV infection. However, in our study no specific target antigens were observed in sera of patients who had recent episodes of HCMV infection.
X-ray irradiation on blood products is a common method to prevent the risk of inducing PTGVHD, and has recently become to be used in Japan. We have tried X-ray irradiation on WB, CRC and PC using HITACHI X-ray Irradiation Apparatus MBR1520R, and studied its practical usefulness. In case of irradiation on blood bags as the minimum dose of 1, 500 rads, these conditions are thought to be practical: 1) the use of 1.0mm Al filter 2) the distance of 550nm from X-ray source 3) irradiation on 4 bags at the same time. But, it has also been noticed that total doses and qualities of X-ray absorbed into blood were different between upper and lower side of the bag. Bloods on upper side absorbed much doses and a wide range of X-ray, on the other hand, bloods on lower side absorbed less doses and hard X-ray. In these conditions, irradiated lymphocytes showed a complete inhibition of thymidine uptake in MLC test, still had 15% of activity in PHA stimulation. The qualities of other blood components have not changed before and after irradition. X-ray irradiation is useful in a routine work of blood center, but problems of proper doses and a uniformity of irradiation are remained to be sloved.
In spite of the use of X-ray irraadiation on Blood Products, few data about its effect on components are reported. We need more informations about a quality of irradiated red cell component. This study shows in vitro changes of irradiated red cell component in WB, WRC and LPRC as the minimum dose of 1, 500 3, 000 and 5, 000 rads. The fact as follows were observed in response to irradiated doses: 1) increased fragility of red cell membrane 2) incresed amount of plasma K and plasma Hb 3) decrease of ATP in WB. 2, 3-DPG, glucose, pH, Ht, Cl and numbers of RBC, WBC and Platelet were not affected by irradiation with doses between 1, 500 and 5, 000 rads. According to these results, followings are recommended: 1) irradiation with 1, 500 rads is a proper method for WB 2) in order to avoid the risk of increased plasma K, WB should be used within 1 week after irradiation, and WRC and LPRC should be used 24 hours after irradiation.
Anti-HCV antibody (anti-HCV) tested by means of ELISA was positive in 33 of 38 (86.8%) hemophiliacs examined, and the mean age of the positive patients was lower than that of the negatives. Positivity of anti-HCV was correlated neither to the types of hemophilia, nor to the presence of HBV and/or HIV infection. High GPT levels could be found more frequently in the anti-HCV positive group than in the negative one. In two of our hemophilia A patients, anti-HCV was converted from negative to positive in 1988, during they were administered with the dry heat-treated FVIII concentrated products. As for the human plasma preparation products, anti-HCV was detected as follows: 41/43 lots (95.3%) of immune globulin, 0/65 lots of albumin, 0/2 lots of heat-treated plasma protein fraction, 2/5 lots of FVIII, 0/3 lots of FIX, and 4/4 lots of fibrinogen products. Those findings show the necessity of checking all of the raw human plasma materials for HCV.
In an attempt to eliminate the hazards of homologous blood transfusions during major orthopedic surgery, 19 female patients donated 1, 200ml of autologous blood before operation. 400ml blood was donated once a week for three weeks before operation and kept in a refrigerator at 4°C. Patients were divided into 3 groups: Group A (5 patients) received 6, 000 units of r-HuEPO three times a week. Group B (6 patients) received 6, 000 units of r-HuEPO twice a week. Group C (8 patients) were controls and did not receive r-HuEPO. All the patients were given ferrous sulfate, 210mg of iron, orally every day from 2 or 3 weeks before the first donation. All the patients were able to collect 1, 200ml of autologous blood. After donation of 1, 200ml blood, hemoglobin levels of each group became 93.5±5.1% (12.7±0.5g/dl), 91.5±1.6% (11.8±0.3g/dl) and 77.6±3.4% (10.7±0.6g/dl) of those at the predonation. Group A and Group B remained almost within normal hemoglobin level. These levels were significantly higher in Group A and Group B than in Group C. The lowest hemoglobin levels after operation of each group were 74.5±8.6% (10.1±1.0g/dl), 71.8±9.3% (9.0±1.0g/dl) and 58.6±7.6% (8.0±0.9g/dl) of those at the predonation. Both Group A and Group B used autologous blood only, while 2 of 8 patients in Group C received 2 or 3 units of homologous blood. During donation any side effects or complications did not occur and after donation none of the transfusion transmitted diseases were recognized. Predeposited autologous blood transfusion is a simple, safe, and cost-effective method to prevent transfusion related reactions and infections, but 1, 200ml blood donation in 21 days without using r-HuEPO sometimes leads the patients anemic and requires additional homologous blood. In this study, 1, 200ml of autologous blood donation was performed safely and easily using r-HuEPO, so it can be possible to perform most of the elective surgery using predonated autologous blood.
A 59-year-old man having arteriosclerosis obliterans (ASO) was associated with the anti-i antibody (anti-i) of IgM class. His chief complaints were multiple painful ulcer formations on his both lower legs and he had been administered with mefenamic acid. Although laboratory tests revealed no findings suggestive of lymphoreticular malinancy nor infectious mononucleosis, elevated anti-EBV antibody titer (EB. VCA IgG 160×), positive direct Coombs' test and possible hemolytic anemia were recognized. So far, only several cases with anti-i have been reported in Japan, all of them having hematological disorders such as malignant lymphoma or Wiskott-Aldrich syndrome. In our case, the mechanism of the anti-i production might be attributable to increased i antigen due to ASO, immunological aberrations caused by mefenamic acid and polyclonal B cell activation due to EBV infection.
Acute heart failure was observed in two patients of cold agglutinin disease with extremely high anti-I antibody. The first patient was 86-year-old man and the second 63 y. o. woman. Raynaud's phenomenon, acrocyanosis, dyspnea, edema and ECG abnormalities such as A-V block, SVPC, VPC and rapid AF appeared after mycoplasma pneumonia. High titer anti-I was detected (×16384 in first p., ×4096 in second p.). These symptoms and ECG abnormalities occured repeatedly, but disappeared soon after warming of the body. The anti-I in the second patient decreased after 6 months, and she recovered. The first patient, however, show 512-1024 titration of anti-I during three years and the symptoms appear repeatedly in this time after exposure of the body in low temperature. Cold agglutinin may cause variable symptoms and blood sluggishness and elevation of blood viscosity. The acute failure of these patients may be due to the changes of these blood stream.
In order to promote rationalization and economization of blood transfusion test, the utility of automated laboratory workstation Biomek 1000 for blood grouping, irregular antibody screening, hepatitis B antigen and antibody screening, etc. of patients' specimens was compared with manual operation. Processing was much faster with Biomek 1000 than that with manual operation in each test. The volume of reagent used with Biomek 1000 was smaller that that with manual operation at blood grouping and antibody screening. The results with both procedures were correlated well in each experiment. Then, Biomek 1000 is very useful with respect to rationalization and economization of blood transfusion test and protection of laboratory staff against infectious diseases.
It is indispensable to determine HBs antigen to prevent hepatitis which may be caused by blood transfusion in some cases. However, since RPHA, RIA and ELISA methods have been performing in past years, they require a skillful technique with intricate handling, while in-hospital blood collection is being conducted in emergency requirements in order to transuse blood into patient without results to be obtained by clinical screening processes. In this clinical aspect, this paper (thesis) reports study with regard to the “LPIA-100” that enables to meet emergency screning for HBs antigen on the basis of (based on) the near infrared latex photometric immunoassay.
Kitaosaka Red Cross Blood Center introduced an apparatus which permits blood donors undergoing apheresis to listen to their favorite music from two loudspeakers placed near their ears and feel vibrations produced by sixteen transducers using low tones less than 120Hz. Both skin temperature and galvanic skin resistance of those donors changed only slightly when this apparatus was not used, and increased when it was used. These results suggested that the donors felt relaxed when the apparatus was used. A study on impressions the apheresis donors received indicates that this apparatus not only gave a pleasant refreshment to the donors, but also made the time required for apheresis shorter than it was, and helped to popularize apheresis. This apparatus also has a merit of preventing vaso-vagal reactions (V. V. R.), a side effect of apheresis. It is therefore concluded that this apparatus is a useful adjunct to donor apheresis.