Dearth of donors, recently becoming intensified, has led to a tendency for blood collection outside of regularly established blood bank installations, which necessitates transportation of preserved blood after collection. We have investigated the effects of mechanical shaking and long distance transportation on the blood.
1) Observations have been made on the possible change during 28 days produced on fresh ACD preserved blood samples by mechanical shaking. No remarkable changes were noted as compared with unmanipulated samples. However, rather remarkable hemolysis was noted during storage when rapid cooling and violent shaking or high temperatures were applied.
2) No significant differences were noted in the effect of long distance transportation among blood samples stored at varying periods of time.
3) These findings indicate that bloods may safely be collected outside the blood bank installations and that these bloods after long distance transportation can be utilized for transfusion without any danger.

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In order to investigate the occurrence of several cases of supposed or suspected infection of malaria after transfusion of preserved blood, the author studied survival of plasmodia in preserved blood in the following experiments, using mainly Plasmodium berghei (Vincke and Lips) in mice and Plasmodium vivax in humans.
1. Plasmodium berghei seemed to preserve its infectivity to mice up to the 3rd day in stored blood after inoculation with anticoagulant Formula B, up to the 2nd day in the same with anticoagulant Formula A and up to the 5th day with simple sodium citrate.
2. The infectivity of P. berghei seemed to depend upon the dose of the pathogens.
3. In using Plasmodium vivax to human for therapeutic purpose, 2 of 4 recipients showed positive symptoms of infection, who received the contaminated blood preserved with ACD Formula B for 4 days. The incubation period in the other 2 patients was 16 and 17 days after the inoculation respectively.

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In the course of a study of the effects of formalin on red cells it was noted that formalin was capable of completely inhibiting the agglutinability of red cells by anti-sera without destroying the blood group antigens of the red cells.
At first, the effect of concentration of formalin and duration of treatment on the stability of red cells was examined. Various dilutions of formalin were made up in buffered saline, the solutions being adjusted to pH 7.3. Five per cent suspensions of washed human red cells were made with the diluted formalin and allowed to stand for varying lengths of time at 25°C.
The mixtures were centrifuged, and the formaimn treated red cells were washed 4 times with buffered saline.
It was observed that with short periods of standing in formalin there was some hemolysis on washing the treated red cells, but after 24 hours no hemolysis was observed upon washing the cells.
Agglutination tests were carried out by mixing dilutions of anti-sera with equal volumes of a 2per cent cell suspension, incubating at 37°C for 30 minutes, centrifuging the mixture and observing for agglutination by gently agitating the tubes.
The agglutinin titers observed with anti-A or anti-B sera and group A or group B red cells treated with various concentrations of formalin for 24 hours at 25°C are given in Table 1.
It is observed that the lower concentrations of formalin decrease the agglutinability of the red cells while the higher concentrations suppress it (Fig. 1).
Serum was absorbed by mixing one volume of washed packed cells with one volume of serum at room temperature for 15 minutes. The results of an absorption experiment with group A or group B red cells and anti-A or anti-B serum are given in Table 2.
It is observed that formalin-treated cells removed the agglutinin, while group B formalin treated cells did not remove the agglutinins from anti-A sera.
When the formalin treated red cells used in the absorption experiment were thoroughly washed, mixed with an equal volume of buffered saline and heated at 56°C. for 15 minutes, the specific agglutinin was eluted.

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In order to ascertain the functional changes of preserved blood, the O₂-carrying capacity of human and rabbit blood in A. C. D. solution was determined at various storage temperatures. The volume of oxygen absorbed in the blood was measured by the use of Yuhu's microgasmanometer and the degree of spontaneous haemolysis in the sample was estimated with Shimazu's spectrophotometer,
The storage temperatures chosen were -5°C, 5°C, 20°C, and 37°C. The blood stored at -5°C was usually not frozen but merely supercooled.
The O₂-carrying capacity of Hb isolated from red cells by freeze-thawing method and stored in A. C. D. solution was also investigated.
The results obtained were as follows: The O₂-carrying capacity of rabbit whole blood stored at -5°C was less affected than that stored at 5°C. The degree of spontaneous haemolysis of rabbit blood stored at 5°C was greater than that stored at -5°C, as the period of storage was extended. In the case of human blood, however, the degree of spontaneous haemolysis of specimens stored at 5°C was less than that of those stored at -5°C, no significant difference in the O₂-carrying capacity between those stored at these temperatures during the whole period of storage (7 weeks) having been observed. The O₂-carrying capacity of human whole blood stored at 20°C or 37°C was affected more remarkably than that stored at 5°C. The O₂-carrying capacity of Hb solution stored at 5°C was not impaired during the 40 days of storage, indicating that haemoglobin is relatively stable in the cold state.

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Clinical and experimental studies on the hematological changes during and after the procedure of extracorporeal circulation, hypothermia and massive blood transfusion were made.
1) The disappearance of leucocytes and platelets occurred during the operations. The sequestration of these blood cells occurred in hypothermia, and the destruction in extracorporeal circulation.
2) Hemolysis accompanying the extracorporeal circulation did not disturb the renal function.
3) Decreased platelet counts were restored in one week postoperatively, but both erythrocyte and leucocyte counts were decreased between 5 to 14 days.
4) Experimentally, the half life of erythrocyte survival was shortened in all cases, being particularly severe in massive blood transfusions. Clinically, the normal half life was obtained in the brain cooling and massive blood transfusions. The shortened half life in the artificial heart-lung pump oxygenator did not completely explain the postoperative anemia.
5) It appeared that the extracorporeal circulation, especially brain cooling, stimulated erythropoiesis experimentally. In clinical cases of artificial heart-lung pump oxygenator, erythropoiesis was stimulated transiently followed by the period of inhibition.
6) Taking the hemodynamic changes into consideration, postoperative hydremia, rather than anemia, seemed to be responsible for the inhibition of the bone marrow function.

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The author measured circulating blood volume, extracellular fluid volume, venous hematocrit value, serum electrolytes, and plasma protein concentration before, during and after the intracardiac surgery by the pump-oxygenator (metal finger pump and DeWall-Lillehei type oxygenator), and studied the changes of circulating blood volume and venous and body hematocrit value before and during the extracorporeal circulation.
The results obtained were as follows:
1) Circulating blood volume decreased immediately after the perfusion in both experimental and clinical cases, in spite of blood transfusion in amounts greater than those of blood loss.
A part of circulating blood is presumed to be pooled in the storage depots in patients or dogs.
2) In experiments with dogs, venous hematocrit values increased during the perfusion, while body hematocrit values decreased slightly. It is believed that these changes were caused by the unbalanced distribution of red cells in the blood vessels.
In clinical cases venous hematocrit values decreased immediately after the perfusion.
3) Fxtracellular fluid volume measured by thiocyanate increased in almost all cases immediately after the perfusion in proportion to the increase of body weight.
4) Serum potassium concentration decreased during and immediately after the perfusion, but the maximum increase was noted on the 1st and 2nd postoperative days.
Serum sodium and chloride concentrations increased slightly during the perfusion and showed decreased level for about one week after the surgery. Extracellular sodium and chloride volumes increased for about two weeks postoperatively.
5) Plasma protein concentration decreased during the perfusion and for about one week after the surgery.
Consequently, it should be emphasized that the administration of saline solution or 5 percent glucose solution must be restricted during and after the intracardiac surgery.

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In 33 mongrel dogs, the author investigated the change of some blood ingredients and blood pressure in groups as divided according to rapid excessive, slow excessive, large hemorrhage-replacement blood transfusion and to mechanical heart and lungs.
1) The changes generally observed in these experiments consisted of hemoconcentration and significant decrease of the solid constituents of blood and impairment of hepatic functions.
2) The circulating blood and plasma volume and hematocrit values were increased with excessive blood infusion, unless associated with liver disturbances. But the volume by which the blood in circulation was increased was less than the volume of the infused blood, probably because various compensatory mechanisms or the mechanism of extravasation were operating in the congested body. Hematocrit was increased in proportion to the speed of infusion rather than to the volume.
3) The plasma protein was increased in value over the corresponding protein infused, the protein fractions showing a reduction in the A:G ratio, without giving any sign of extravasation.
4) Of the formed elements of blood, the red blood cells were increased in number, though less in number than that of infusion, in those cases in which the method of infusion used caused serious mechanical impairment. The white blood cells and platelets were remarkably decreased in number.
5) The other changes inevitably produced in any massive transfusion were prolonged thrombin time, reduced pH of blood, and fibrinopenic condition.
6) Liver impairment, hemorrhagic tendency and other disorders produced are all curable sooner or later unless there are some complications, such a s liver disturbance developing previously. Transfusion in the presence of liver impairment and the effect of ACD as a preservative were reevaluated.

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