The growth curves of three experimentally contaminated strains of Pseudomonas and one of Aerobacter aerogenes in banked blood were shown.
During first 24hrs. after inoculation, the viable number of bacteria decreased remarkably on account of bactericidal activity of serum, but surviving cells started again to grow during the following 24 to 48hrs., the growth being completed in 7 to 14 days.
Seventy-one per cent of 40 strains grew abundantly in banked blood when incubated at 5-6°C, while only one strain among other water bacteria (21 strains of Achromobacter, Alcaligenes and Flavobacterium) multiplied under these conditions.
Therefore, it is obvious that pseudomonas is the most dangerous contaminant among water bacteria.
In order to explain the mechanism of shock syndrome developing after the transfusion of contaminated blood, the effects of intraveneous injection of washed bacterial cells into experimental animals were investigated.
A remarkable depressor action was demonstrated when the cell suspension of either Aerobacter or Pseudomonas was injected. On the other hand, the action was not significant with Micrococcus. Should an accident occur, it is necessary to isolate the contaminated bacteria from the patient or after death.
The author demonstrated that if the contaminant was coli-aerogenes group organism, the causative agent was easily isolated from the human materials. However, the isolation was not successful in the case of pseudomonas, because the water forms of pseudomonas were destroyed in an early stage of injection.

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For the purpose of investigating the infectivity of virus of Japanes B encephalitis, added to human blood or serum, the author made experimental observations on the relation between the infectivity and the conditions under which serum was preserved.
10^2.2 ID₅₀ of virus of Japanese B encephalitis were added to pooled human plasma in a bottle, and stored at 4°--20°C., or in a lyophilized state for 5 months.
Each sample was inoculated intracerebrally at monthly intervals into eight mice for testing the infectivity of the virus remaining in the samples.
The results showed that the infectivity of virus in serum stored at 4°C. or in a lyophilized state disappeared one or two months later, and that in serum stored at -20°C. remained for 3 months but disappeared after 5 months.

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The author investigated dynamic changes in several components of whole blood during storage at 4 C. The results are summarized as follows:
1) Glucose and inorganic phosphate in whole blood decreased rapidly during the first four weeks of storage, but lactate increased in value. Relation between glucose and lactate concentration was not stoichiometric.
2) Citrate in whole blood showed no change during storage.
3) Pyruvate accumulation was shown to rise rapidly after four weeks of storage.
4) Oxidation-reduction potential of the pyruvic-lactic acid system was lowered gradually through 4 weeks, and rose slowly thereafter,
5) Carbon dioxide in whole blood showed no increase.
6) There was no change in pH of whole blood during storage.

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The author observed the effects on the synthesis of phosphate ester in ACD blood treated with adenosin or ribose. The results are summarized as follows:
1) In ACD blood treated with adenosin, inorganic phosphate decreased whereas esterified phosphate increased.
2) In ACD blood treated with ribose, however, inorganic phosphate increased as esterified phosphate decreased in value.

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Since carbohydrate metabolism is closely related to phosphorus metabolism, the author investigated the transport of inorganic phosphate through the erythrocyte membrane with the radio-isotope tracer technique using ³²P. The results may be described as follows:
1) The elution of ³²P from labelled erythrocyte was identical regardless of elution media.
2) The elution of ³²P into phosphate buffer was inhibited by the addition of 0.01M monoiodoacetate, but was not inhibited by 0.01M of sodium fluoride.
3) Uptake of ³²P declined with the preservation of ACD blood. The author suggested that the mechanism of ³²P uptake was enzymatic.

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Since large amonnts of blood have readily become available for blood transfusions, such undisirable complications as the development of hemorrhagic syndrome are being increasingly noted in the recipients. There are many possible considerations in regard to the etiology of this side-reaction, such as, the dilution of the clotting factors by the infused blood, in which labile factors have lost their activities during storage, the decrease of platelets in the recipient's blood, the intoxcation by citric acid used as an anticoagulant, and the acceleration of fibrinolysis.
Of these consideration, several observers have pointed out the activation of the fibrinolytic system during the storage of blood. Therefore, it seems necessary to perform a systemic analysis of plasminogen, plasmin, and antiplasmin in the stored blood.
However, there is no evidence of the conversion of plasminogen into its active form during the storage of blood at 0-4°C, and no influence of anticoagulants usually employed in blood bank, such as, ACD solution, EDTA-Na₂ solution, or 3.8% citric acid solution, for the activities of the fibrinolytic system with the exception of the very weak antiplasmin action of EDTA-Na₂ solution, according to the observations reported here.
Furthermore, it is also obvious that the platelets represent the antiplasmin activity and their number decreases rapidly by storage. It follows that in the presence of the enhancement of fibrinolysis in a patient receiving massive blood transfusions, the activity of the stored blood may be relatively weak in regulating the fibrinolytic system in the recipient because of the reduced number of platelets.
On the other hand, while ε-aminocaproic acid shows a potent inhibitory action on plasmin, no effect can be detected against the activation of plasminogen to plasmin as determined by the precipitation method using 1M. NaCl (pH 2.0). It will be reasonable, therefore, to administer ε-aminocaproic acid for arresting bleeding tendency due to fibrinolysis occurring after blood transfusion, although it is not necessary to add ε-aminocaproic acid to the anticoagulant solution for blood banking.

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Purpose of this experiment was to find a quantitative indication for bactericidal activity of citrated whole blood in which was represented at 37°C. When the experimental materials were removed from the incubator at 37°C to it at 4-6° C, and then cultured, the logarithmic growth curve of surviving bacteria in this materials was parallel to the curve of survival bacteria in bouillon under the same experimental condition. The experiments showed that the logarithmic growth curve of surviving bacteria was regularly constant, and the proportion of the number of bacteria counted in the beginning was almost parallel to the distance of the position between the curves of these bacteria. It was found that the bactericidal activity might be expressed on the number of days (L-Index) from which the distance was calculated by a general arithmetric formula for computing a method of relative potency. The computated standard was found to be a curve of logarithmic surviving bacteria-growth in bouillon. The computated distance was represented by the diference of the positions between the growth curve in materials and in bouillon. The potency of bactericidal activity in test materials could be expressed by the number of L-Index. Studies on the mechanism of bactericidal activity in human stored blood was investigated by this calculating method. The results of this studies pointed out that this method of the expression on bactericidal activity could be used in practice.

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The method for producing anti-human complement antibodies previously reported by Matuhasi and Usui was further investigated. The immunization of rabbits was carried out as follows: Egg white saline to 1:5 and antiegg white rabbit serum were mixed together with fresh human serum at the serological optimum ratio. After the precipitate was centrifuged, the complement titer of the supernatant was titrated. It was found that the titers were one tenth or one fortyfifth of the initial titer of the fresh serum, in other words, about 60 to 100 units of complement were fixed to the precipitate.
The three times washed precipitate was injected into rabbits subcutaneously twice a week for 3 to 4 weeks. Ten days after the last injection the rabbits were bled and the obtained antisera were tested by direct and indirect Coombs tests, the Ouchterlony technique and the analysis by the field reaction of the ring test. It was confirmed from these techniques that the antisera contained mainly anti-non-γ-globulins, in particular anti-complement antibodies.
The anti-human globulin antisera containing mainly anti-γ-globulin were prepared by immunizing the rabbit red blood cells treated with the heated supernatants which human complement had already been absorbed with the precipitate. The rabbit red blood cells were expected to be coated by human heteroantibodies belonging to the γ-globulin and to fix only small quantity of complement. In fact it was observed by the various serological techniques that the immunized rabbits mainly produced anti-γ-globulin antibodies.
These findings would lead to the conclusion that anti-human complement antibodies can be easily produced by immunization with antigen-antibody complex which is able to fix human complement.

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