As a part of our fundamental studies on paper electrophoresis we here present our views regarding selection of the two methods of paper electrophoresis, the Grassmann and the Flynn method. The filter paper used was Whatman No. 1, the stain, Amido Black 10 B.
Ten essentially normal human sera were used, and the results from both methods were almost identical, although differing in minor details. It is these minor differences that we wish to consider here.
(1) With the electrophoretic time being the same for both methods, the length of the electrophoretic migration was longer for the Grassmann method than for the Flynn method. This may be due to the movement of the buffer toward the center of the paper, as a result of the evaporation from the surface of the paper.
(2) The position of γ-globulin with the Flynn method was always on the negative side of the line of origin (electroosmosis), but with the Grassmann method it was consistently on the positive side. This, also, we consider is due to the flow of the buffer to compensate the evaporation loss.
(3) The protein adsorbed at the line of origin was more marked for the Flynn method than for the Grassmann method. We are now considering a way to eliminate this defect in the Flynn method. In order to observe the tailing of the albumin, therefore, the Grassmann method is more convenient.
(4) The reproducibility of both methods, as compared to free electrophoresis was not very good, owing to the evaporation from the paper and also to the sagging of the paper in the Grassmann method.
(5) From the above it is concluded that although the Grassmann method is more popularly used, the Flynn method when properly handled is far from being inferior to the former method.

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As a part of the study to learn the mecanism of the Miyamoto-Sasaki Cancer Test, effect of ions on the crack number of blood serum was studied.
“Crack number” means the grade to represent the results of the Miyamoto-Sasaki Cancer Test. (See Fig. 2)
Generally, cations make the crack number of the Miyamoto-Sasaki Cancer Tests smaller, probably because they accelerate the heat coagullations of blood sera.
Those cations which have valences of 1, e. g. Li⁺, Na⁺, and K⁺ with their Mol/10, and those which have valences of 2, e. g. Mg⁺⁺, Ca⁺⁺, Ba⁺⁺, Zn⁺⁺, Fe⁺⁺, and Mn⁺⁺, with their 3Mol/5, 000-3Mol/10, 000 make the crack number remakably smaller.
CaCl₂ of 6mg/dl and MgCl₂ of 1.5mg/dl are enough to make the crack number 1 grade smaller than before they are added to the serum.
Concerning the problem whether the changes of Ca⁺⁺ or Mg⁺ concentration in the living body can be a factor which has a hold upon this Cancer Test or not, author has not got a conclusion yet. But it is a remarkable fact that above-mentioned small concentrations of Ca⁺⁺ and Mg⁺⁺ have effects on the crack number of the Miyamoto-Sasaki Cancer Test.
Anions which have valences of 2 or 1 seem to have not any effects. Anions valences of 3 have reverse effect in some grade.

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The meaning of electrophoretical analyses of protein of vegetable tissues in the study of vegetable physiology is considered to be very important, and we prescribed the conditions of its electrophoresis at first.
That is, we proposed newly the mixed buffer of K-phosphate and Na-borate, and fixed in details the conditions of electrophoresis concerning this buffer.
Then under the fixed conditions of electroporesis, we sought the interrelation between the electrophoretic protein of vegetable tissues and the physiological phenomena.
The more vigorous the functions of leaves or seeds, the more complex the electrophoretical patterns of the extracts of these tissues.
Thus, the existence of close interrelation between physiological phenomena and electropholetic protein was shown, and, moreover, the dynamic character of electrophoretic protein was inferred by the results of autolysis.

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Although mammalian red cells, in general, have biconcave discoid forms, they change readily their shapes, however, in saline solution and especially in the “oligolytic concentration”, 0.4mol, assume promptly crenated spheres. Furchgott and Ponder reported that the biconcave discoid erythrocytes in the physiological saline soultion change the shapes easily by changing the pH of the medium and become immediately spheric in alkali.
The authors investigated the disk-sphere transformation of the red cell in various concentrations of saline by adding sodium hydroxide and studies the shape change from the view-point of the light absorption of the erythrocyte suspension.
The spheric transformation of the 0.032% erythrocyte suspension by the addition of alkali, especially 1/1000N to 1/12000N sodium hydroxide, was studied spectrophotometrically. The shape change of the red cell could be detected by the light absorption curve as in the case of the solution of the neutral salt.
1) The red cell assumes readily the complete spheric form by the addition of sodium hydroxide in the lower concentration than the oligolytic concentration and the more readily with the decreased concentration of saline. At the same time, the light absorption curve of the erythrocyte suspension shows an decrease in short wave length.
2) The decrease in the light absorption curve in short wave length, however, is small with the decreased concentration of saline even if the red cells are the spheres of same size, and at the same time the effect of the absorption bands of hemoglobin contained in the red cell is observed a little.
3) When sodium hydroxide is added to the erythrocyte suspension, the red cell is transformed spheric and hemolyzed in several minutes or hours according to the concentration of sodium hydroxide. If the concentration of sodium hydroxide is high, the red cell assumes again the biconcave discoid form just before hemolysis occurs. The phenomenon is also related to the concentration of saline as a dispersion medium and the disk transformation occurs by the more diluted sodium hydroxide with the decreased concentration of saline, but occurs hardly with the increased concentration of saline. When the red cell becomes again biconcave discoid, the light absorption curve increases in short wave length and the obvious absorption bands by the effect of hemoglobin are obseved in 420, 540 and 580mμ.
4) In the case that the dispersion medium consists only of saline solution, the light absorption of the erythrocyte suspension shows in the oligolytic concentration, 0.4mol, the remarkable maximum in 720mμ and minimum in 420mμ. When sodium hydroxide is added to the dispersion medium, the light absorption in the lower concentration than the oligolytic concentration shows remarkable change, which is decrease of the light absorption in 420mμ and increase in 720mμ. In this case the maximum of minimum is not obvious.
5) And the change of the light absorption in the lower concentration than the oligolytic concentration is more conspicuous with the stronger bases in the same normality.

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[in Japanese]

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