1. Objective and Procedure
Fresh red blood cells are to be frozen either slowly or rapidly with glycerol for preservation as viable cells in a frozen medium. From the viewpoint of deglycerolizing such preserved viable erythrocytes there have been introduced three different techniques, viz., the reversible agglomeration (Huggins' method), the batch washing and the continuous centrifuge procedure.
We conducted a study attempting the evaluation of the continuous centrifuge technique for deglycerolizing preserved red cells, using a Hemonetics Model 15 apparatus, in comparison with Huggins' method as to effects on osmotic pressure, free Hg concentration, erythrocytic resistance, electrolyte balance, 2, 3-DPG and ATP levels and recovery rate of viable cells.
2. Results
As compared with Huggins' procedure, the continuous centrifuge method produced rather inconsistent results for which the rates of red cell and washing medium injection appear to have played an important role. In case of the washing medium being introduced into the apparatus before erythrocytes it would be probable that sufficient washing of red cells could hardly be accomplished. The failure in obtaining constantly optimal osmotic pressure, erythrocyte resistance and electrolyte balance with the subject procedure has thus been conjectured to be due to insufficient washing of preserved cells.
3. Conclusion
The continuous centrifuge seems to be valuable method provided it be performed attentively insomuch as the glycerol concentration employed is fairly low and as electrolyte solutions are used for washing the cells.

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The preserved blood for transfusion can be kept in a refrigerator at 5°±1°C to 21 days. However, it is not sure that the temperature is being kept all the time at this range because of transportation, operating room procedures or disturbances in the refrigeration systems.
The effects of temperature were determined at 5°, 21° and 37°C from 0 to 48 hours on the properties of red cell membrane resistance, sodium and potassium content in plasma and ATP and 2, 3-DPG contents in blood.
At 37°C the changes are so big that 5 hours correspond to a week storage of the blood and 8 hours to 2 weeks of storage. 8 hours at 21°C correspond to a week of storage. The biggest change was shown on the decrease of 2, 3-DPG content.

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Intravenous administration of human IgG frequently causes hazardous side effects owing to the molecular aggregation of IgG followed by fixation and activation of complement in vivo. To avoid these unwanted effects, acid treatment are in practice. Chemical modifications of IgG also decrease anticomplementariness, and treatment with β-propiolactone by Stephan et al. has an advantage of longer survival time in vivo than that of enzyme treatment because it gives little change on the molecular weight. However, it is known that chemical treatments such as amidination, benzylation or carbamylation reduces antibody titer. In this study we investigated the anticomplemetary activities and the antibody titers of two lots of commercial preparations of human immunoglobulins treated with β-propiolactone (βP-IgG) to SLO, influenza, measles and diphtheria toxin in comparison with non-treated human immunoglobulins (FII) as well as the pepsintreated immunoglobulin preparation (pep-IgG).
The antibody titers of βP-IgG were almost as equal as those of pep-IgG or FII, although small variations probably due to differences between the original titers of materials (pooled plasma) exist.
The anticomplementary activity of βP-IgG was 12.2 CH50%/50mg which revealed to be about a quarter of that of FII (43.1 CH50%/50mg) and within Kistler's critical value (0.6%/mg), however in comparison with pep-IgG (2.3 CH50%/50mg), it showed 5 times as much anticomplementary activity.
Marked increase in anticomplementary activity was noted during storage of the preparations at 37°C for 4 months, although antibody titers were nearly unchanged.
On the other hand the anticomplementary activity increases less than half in comparison to the above state during storage of the preparations for the same period at 4°C.

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