The previous report issued from this laboratory has indicated that HIM test developed by Bolin was effective for the blood screening method. However, about 40% of blood donors were shifted out by original HIM screening test with using a serum dilution of 1:5 and this result showed that it was a little difficult in practical application because of short supply of donors blood for transfusion.
So, this study was made to seek for possibility of obtaining the more practical modification and its application of original HIM donor screening test.
The results obtained were summarized as follows.
1) HIM quantitative test up to the titer 1:40 was performed on the 1276 cases of normal donor serum. It was found from the results of these tests that approximately 86% of the donors had a titer 1:10 or less when the distribution of antibody level was inspected in normal donor.
2) The each donor's blood was classified to two groups, i. e. Positive ones and negative ones, by the HIM test with using a serum dilution of 1:20 as a basis of donor screening. It was found that the incidence of post-transfusion liver dysfunction lowered significantly in the group of recipients to whom HIM negative blood was given than in the group of these HIM positive blood recipients.
3) Before the blood transfusion, the patients to be transfused were screened by HIM testt with 1:5 serum dilution, and were classified into two groups, HIM positive patients and the negatives. After blood transfusion, the patients of two groups were observed for 6 month as to the difference in the incidence of posttransfusion liver dysfunction. It was found that the incidence of post-transfusion liver dysfunction lowered in HIM positive patients group significantly than in the negatives.
From the result of these observations, combination of donor screening by HIM test with using a serum dilution of 1:20 and pre-transfusion HIM antibody screening of patients is suggested to be very effective and practical in preventing measure from post-transfusion hepatitis.

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These studies were undertaken to clarify some causes of posttransfusion hemoglobinuria of frozen blood processed by modified Huggins procedure. For these purpose, the membranous characteristics of the fresh ACD, the storaged ACD and the frozen red cells were morphologically classified into four types by use of the low power (5000 ×) electronmicroscope. The criteria for the classification are basedonn theentire shapes, the thickness of the outer layers and the surface conditions with or without holes of the membranes.
The following results were obtained.
1. The type I is considered as the most juvenile one, and the type the most aged. The type I and II are the intermediates.
2. These aging-like phenomena of the red cell membrane in vitro were occured and observed either during the preservation in 4°C as the ACD blood or during the processing of frozen blood. The type I transformes to the II the II to the III and the III to the IV. The final one is hemolysed. These aging-like phenomena might be caused by processings of freezing, thawing, washing and resuspension, however it is still in controversy which one is the most effective factor for the membrane damages. The aging-like phenomena occured remarkablly when the 14th or the later dated ACD blood after collection were used for the frozen blood.
3. Considering the above results and the clinical experiences of frozen blood transfusion of 378 units for 93 patients, of whom 7 cases showed hemoglobinuria, the more juvenile red cells will be the tolerable and preferable for the processing, and the deadline of ACD blood to be used as the frozen blood showed be settled on the 7th day after collection. And, the effective volume of cryoprotective agent should, of course, be added to the red cells.
4. Frozen in a glass bottle for japanese one unit transfusion which has a limited capacity of 230ml, posttransfusion hemoglobinuria might be caused from shortage of Huggins 8.6 molar glycerol solution to be added into the bottle, althought the fresh red cells in the 7th day were used.

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