The active guinea-pig anaphylaxis was studied by Insulin as antigen. The results were as follows:-
1) Insulin made in Japan as well as foreign made, when injected to the sensitized guinea-pigs once again, causes light anaphylaxis. However, guinea-pigs sensitizcd by fish Insulin, injected animal Insulin, are negative, and guinea-pigs sensitized by animal Insulin injected fish Insulin, are negative too.
2) Guinea-pigs sensitized by inactive Insulin, when injected once again by active Insulin, cause light anaphylaxis.
3) Guinea-pigs sensititized by Commercial Insulin, when crystal Insulin is injected, cause light anaphylaxis.
4) Hyper-glykamie was caused after the second injection as mentioned above.

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Complement-fixing antibodies were found from rabbit sensitized by simple injection of Insulin (fish Insulin & animal Insulin), without hitherto-used horse serum etc.
1) By the cross reaction between fish Insulin and animal Insulin, Insulin complement-fixing antibodies were found after one hour of fixing antigen, antibodies and complement at 37°C. and prolonged 24 hours at 3°C; which were never found by Kolmer method.
Insulin, as its molecular weight is small, takes a long time to react Insulin complement-fixing just as the case of Virus.
2) Insulin sensitized serum is positive, when complement-fixing reaction is made using extracts of fish and beef as antigen. This shows that the serum contains foreign matters besides Insulin antigen.
3) The same results were obtained by using crystal Insulin as well as commercial Insulin.
4) Insulin complement fixing antibodies last 3 or 6 months, and strongest reactions are found 7 days after the sensitization.
5) Complement-fixing antibodies were found by a cross reaction rather than by serum of patients treated by Insulin.

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Rabbit was immunized by fish- and bullinsulin, and the complementfixation has been examined by Ogata's antigen antibody dilution method (the insulin complementfixation is succeeded when the fixing time of antigen, antibody and complement is to be acted during one hour on 37°C and after that 24 hours on 3°C).
1) There is some difference in Ogata's equivalent zone on the insulin complementfixation between the insulin of the same kind used in immunity as antigen and other kind insulins (that is, cross reaction). Besides, it is moved according to the immunity process, and it is almost 16 unit.
2) The title of the antibody is 1:8160 in the insulin of the same kind and 1:40 in the cross reaction.
3) The title of the antibody of the complementfixation to make the examination antigen, when the extracted fluid of the same animal protein produced the insulin is 1:1020 in the same serum.
4) The title of the antibody on the cross reaction about the insulin immunized rabbit's serum which was absorbed with the extracted fluid of the same animal protein has produced the insulin is almost equal to one before absorption.
5) The title of the antibody by the extracted fluid of the animal protein produced the insulin of the insulin immunized rabbit's serum which was absorbed by the other kind insulins, (it is dif-ferent from the insulin used in immunity) is almost equal to before.
6) As the insulin will be analyzed with electrophoresis, it will be able to separate the insulin protein and impure proteins.
7) The insulin antibody has been also declared by the insulin complementfixation about the crystal insulin immunized rabbit's serum, too.
As mentioned above, the insulin protein has the immunity and the insulin preparats contain some impure things, which causes impure protein antibodies with the insulin protein antibody.

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An antigen-antibooy reaction was repeated for 6 months long by intravenous injection of 4 times diluted cow serum with phosphatide and by injection of α-streptococci in paranasal cavity of rabbits. They were let free from any procedure for more than 1 month to 8 months. Thus the chronic cerebral local anaphylactic rabbits were made. The Ch-E activity in each motor cortex, Ammon's horn and nucleus lenticularis was measured. The results were as follows:
1) Ch-E acitivty in each rabbit's brain was ranged in the order of the motor cortex (highest), Ammon's horn and then nucleus lenticularis.
2) In the chronic cerebral anaphylactic rabitbs, Ch-E activity in the motor cortex and Ammon's horn was much higher than that of normal one, and in the nucleus lenticularis it tended to increase slightly.
3) The increase of Ch-E activity in each part showed no fluctuation even during long time without any management.
4) The increase of Ch-E activity in each part was not influenced by the kinds of sensitizing antigen or of the methods of sensitization.
5) The permanent increase of Ch-E activity and abnormal accelaeration of brain metabolism in rabbits brains were observed by the repeated reactions as above-stated for a long time It would appear that, the chronic cerebral local anaphylactic rabbits were given an experimental epileptic arrangement.

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The free aminonitrogen in the brain with CCLA was strikingly decreased in comparison with that of the normal, and no difference was observed between the kinds of antigen or the methods of sensitization.
The decrease of free aminonitrogen was not influenced by protraction of the period without procedures after the last anaphylactic reactions, and the decrease was seen to be longstanding, when these very slight anaphylactic reactions were repeated for a long time. This is considered to be due to the abnormal brain metabolism in the state of an experimental epileptic arrangement.

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No striking difference of KEG granules was observed in the rabbits brains between the normal and the chronic cerebral local anaphylaxis KEG granules were not influenced by the kinds of antigen methods of sensitization and the length of period without any procedure after the last anaphylactic reactions.

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After the removal of various centers of the motor cortex in dogs, investigations were made; i.e. The problem of exchange or migration of the motor centers proved by electric stimulation, the function of compensation, implicated arrangement of the nerve cells of the different centers and the compensation between both hemisphere.
1) In the remaining motor cortex a phenomenon of exchange of the motor centers was observed. The migration of the area for the hind limb (H. A.) into the area for the fore limb (F. A.) in case of the removal of H. A. and the mixed area (M. A), was more frequently observed than that of F. A. into H. A. in case of the removal of F. A and M. A. But no migration was observed when all of these F. A., M. A. and H. A. were unilaterally or bilaterally removed.
2) Recovery from paralysis of the fore limb in case of removal of F. A. and M. A. came later than that of the hind limb in case of removal of H. A. and M. A. And in case of the removal of all of these F. A., M. A. and H. A. the recovery of the hind limb came a little earlier than that of the fore limb.
3) When F. A. and M. A. were removed, a slight paralysis of the hind limb was mostly observed in addition to the paralysis of the fore limb. When the H. A. and M. A. were removed, however, the paralysis of the fore limb could hardly observed. This fact was also observed in cases of bilateral removal.
4) After unilateral removal of the motor cortex, when the paralysis of the according limbs had recovered, and then the motor cortex of the contralateral side was removed, there was seen paralysis of the according limbs as well as a slight paresis in the other side.
5) From the facts as above-stated, the implicated arrangement of the nerve cells in F. A. and H. A. could not be considered to be equal and the cells for the hind limb extended more frequently into F. A. than those for the fore limb into H. A.

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When overweight is added to normal bone a relation forms with its transformation. Therefore, the author made experiments on normal adult dogs by resecting parts of their radial bones and giving overweight to its ulna to see what attitude it took.
1) The main X-ray findings were the decrease of physiological curve of the bone and the spindle shaped periostal callus at the part getting the greatest overweight.
2) It was concluded from the distribution of stress and strain in the inner part that the part producing future dynamic callus was the same as the place receiving maximum stress before its thickening. Stress as well as production of callus seen in the compressed part of the concaved side was much greater than that of the retracted part of the convexed side. Stress appearing in the bone after its thickening was generally reduced comparing to its previous state and stability was formed. Comparing to normal bones decrease of the curve helped in reducing stress.
3) Changes seen in nutritional arteries by arteriography showed complicated distribution and fine shadows. Formation and dilation of new vessels were also seen histologically in dynamic callus and bone.
4) Histologically it was mainly the development of periostal callus. There was dilation of Haver's canal and its passage was irregular with its inner vessels dilated and showing hyperemia. Hyperemia, new development and intrusion of vessels were obvious also in the callus.

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Out of the certain experimental results obtained in the isolation as well as synthesis of Thymolindogenide, which has been undertaken to determine certain products of Jolles's reaction in vivo, I could arrive at the following conclusions:-
1) For the first time in our literature, I have succeeded in isolating certain color substances of Jolles's reaction, in the form of a red prismatic crystal whose melting point has proved as 234°237°C.
2) The red prismatic cystal that thus has been isolated proves just the same in its chemical sense with the pure synthetic Thymolidogenide (that has its melting Point at 238°240°C).
3) Color substances which I get from Jolles's and Jolles-Takeuchi's reactions have proved one and the same, both being Thymolindogenide.
4) Those Thymolindogenides which I myself have isolated as well as synthesized have higher melting points respectively, compared to Thymolindogenide synthesized by Jolles, by 16°C or 20°C; moreover, dissolve themselves into crimson when they should have met with glacial acetic acid.
5) In an isolation test in which Jolles's or Jolles-Takeuchi's reactions were adopted, we could observe Indigo and certain non-crystal brown pigment, to be extracted simultaneously into the chloroform, besides Thymolindogenide.
6) In this, I have mentioned the synthetic method of producing the Thymolindogenide out of scanty amount of stuff.

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1) Laying emphasis on chemical equation of Jolles's reaction, and systematically examining those causes for mismeasurement happened during urinal determination of indican, I was enabled to find out certain new microestimating method for indican, which would prove somewhat satisfactory.
2) As indicated below, the normal value of urinal indican estimated by new method, have acknowledged certain marvelously high values that have far superseded those theories hitherto known. However, I am to recognize the Principle No. (2) i.e., the quantitative value for indican that was considered to have as its object the colour reaction against the fuming hydrochloric acid, which could purely has enabled to induce it solely into the Thymolindogenide; while, Principle No. (1), i.e., the quantitative value which has for its object the original color of Thymolindogenide, should be recognized as total Indican value, i.e., that was enabled to be induced into every kinds of Indogenide including the very Thymolindogenide:
(1) Normal amount (mg) of excretion Within 24 hours.
highest 158.0 (Principle No. (1))137.0 (Principle No. (2)),
lowest 42.5 (Principle No. (1))29.8 (Principle No. (2)),
average 98.0 (Principle No. (1))77.0 (Principle No. (2)).
(2) Normal density mg % highest 22.7 (Principle No. (1))19.7 (Prnciple No. (2)),
lowest 2.5 (Principle No. (1))1.8 (Prnciple No. (2)),
average 9.3 (Principle No. (1))7.5 (Prnciple No. (2)).
(3) Misestimation in quantitative value shown by Principle No. (2) to that of Principle No. (1) is estimated as+2.2-43.0%; average, -26.7%.
3) Various stimulants to give rise to mismeasurement on the side of indican at its quantitative determination that have been able to investigate, were as follows.
(1) The condensation between the thymol and indoxyl in the urine increasing at a gradual ratio, and the quantitative value shows a proportional increase as time for reaction lengthens; yet, if the reagent due to Takeuchi (Estimating method (1)) should be used, it may come to an end after two hours' reaction.
(2) The descent in estimation owing to a shortening of reactionary time may roughly be ascribed to the fact that while the condensation between the thymol and indoxyl remains incomplete, the formation of indigo undergoes certain stimulas.
(3) If one might consider the certain of Jolles's reaction as a principle for indican estimation, it would be told as very unsatisfactory: in fact, even after 2 hours of reaction, already it has been seen to have given rise to so marked a difference as -51-71%. Altogether, the chief cause will be pointed out as the incomplete oxidation owing to Obermayer's reagent.
(4) Order in adding chloroform according to Endo's method, assists to formulate Indigo, but in the same degree affects to decrease to formulate the Thymolindogenide; so that, in 2 hour's reactionary course, it brings about already -20% misestimation.
(5) Endo's standard solution, though excellent in its tint, its titer value amount even to 3 or 3.6 times larger than its theoretical value, being remarkably dense. This may take place, perhaps, when he decided on titer, Mr. Endo employed for standard the indican value estimated by Takeuchi's method.
(6) For an urinal oxidizing agent between the thymol and the indoxyl I might mention that of Mr. Takeuchi as best of that kind. The oxidizing agent indicated by 1% potassium chlorate, 3% hydrogen peroside, 15% ammonium persulphate, 4% chlorinated lime, 10% copper sulphate, under the same conditions, only cope with 1/2 power proved by Takeuchi's, at most. Whereas, sodium nitrit cannot stand to be used because apt to produce some nitrosothymol.
(7) The precipitation rate due to precipitator attributed to urinal indican proves different according to the kinds as well as volume of precipitators.

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1) Based on the systematic experimental ground about explanatory tasks for Misestimation of indican, I have invented a new microestimation method for indican found in the blood.
2) Normal value of blood indican estimated due to new method for superseded those various theories hitherto proclaimed, reached so high as 0.261 mg% at highest, while proved at lowest 0.099 mg%; average, 0.190mg%.
3) Complete estimation of blood indican is difficult by using of most estimation method for blood indican in which Jolles's reaction has been adopted as principles.
4) The necessity of re-examination for those various achievement as to clinical significances of blood indican thus clarified hitherto, by using new exact estimation method, has become urgent

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In the recent several years, it is necessary to use the preserved blood in the surgical procedures because of the reasons that the dvelopements of surgical techniques have demended the great amounts of blood transfusions. But it may be said that the preserved blood transfusion has many unknown clinical problems, even its clinical usefulness. The author studied on the relation of hemolysis of preserved blood to untoward reactions of transfusion in Part I of this studies.
1.On the clinical investigations of hemolytic reactions by preserved blood transfusion: The hemolytic reaction is the most important and severe untoward reaction. In this reaction, the patient shows the so-called hemolytic shock and, in the latter, stadium, has the crisis due to renal failure. The causes of this reaction, which shows the destruction of blood cells in the patient, may be said in (1) incompatible blood transfusion, (2) massive transfusion of O-type blood and (3) transfusion of hemolysed blood.
The statistical analysis of the untoward reactions due to preserved blood transfusion in the Tsuda's Surgical Department shows that 80% of untoward reactions are fever or allergic ones and 0.8%, 14 cases, are hemolytic reactions. 6 cases of these hemolytic reactions are occurred in incompatible blood transfusions and other 8 cases are seen in compatible blood transfusions. The main causes of latter 8 cases have the intimate relations to the hemolysis of preserved blood But, it is remarkable that the clinical severity of this reaction is not identified the grade of hemolysis or the quantity of incompatible blood transfusion and is in the more intimate relations to the patho-physiological states of the patients.
2. The grade of hemolysis and the untoward reactions; There are seen no definite intimate relations between the grade of hemolysis, ocurrences of untoward reactions and the kinds of untoward reactions.
3. The patho-physiological states of the patients and the untoward reactions; The author classified the next four groups in the meanings of patho-physiology. (1) nearly normal state, (2) anemia, (3) hypoproteinemia and (4) dehydrated state. In the latter 3 groups there are shown the untoward reactions in the ratio of over twice numbers than the group of nearly normal state. The allergic reactions are seen in hypoproteinemia group in the most high ratio. On the contrary, the fever and hemolytic reactions are often seen in the groups of anemia and dehydrated state.
4. Experimental studies on the hemolysis of preserved blood: The atuthor has the experimental studies on the hemolysis of preserved blood, especially on the influences of preservation days, transportation, shaking, filtratron, temperature, mixtures with many other physiological transfued solutions etc.. All over mentioned mechanical procedures show the unexpected important influences on the hemolysis of preserved blood, and especially show the marked hemolysis in the longer preservation days. These hemolytic tendencies are seen most intense in the duration of 1 week of preservation.
In the conclusions of these clinical and experimental studies, there must be said that it should be taken the most careful handling in the presereved blood transfusions even mechanically as well as criticizing of hemolytic states of preserved blood itsself and shonld use the preserved blood of duration of within one week to the poor risked patients.

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There has been found by author that the preserved blood transfusion results somewhat remarkable pathological changes in the renal functions without any evident clinical signs. In the patients of hypoproteinemia, anemia and dehydrated state there are seen temporary decreasings of urine flow after transfusion. This tendency is more markedly in the various kinds of untoward reactions. Thus the author studied on the pathological changes of renal functions by renal clearance method.
1. Type I of patho-physiological changes in renal functions: This type of patho-physiological changes did not show any kinds of Oxford shunting and is seen in the patients of nearly normal state or anemia without untoward reactions. In this type there was seen the mechanism to maintain the glomerular filtration-function to compensate decreasing of renal blood flow which was resulted by dilatation of extrarenal blood vessels due to blood transfusion. In the other hand, there are seen, in the meaning of humoral mechanism, early adrenalin effects or early antiduretic hormon effects and late DOCA effects or decreasing of mineral excreations.
2. Type II of patho-physiological changes in renal fonctions: This type means the presence of Oxford shunting I in slight grade and is seen in the patients of hypoproteinemia and dehydorated state without untoward reactions. The disturbance in the course of inactivation of antiduretic hormon in liver. This disturbance as well as the decreasing of cortical reactions in epinephral glands results the abnormal renal circulations as Oxford shunting, even it is temporarily.
3. Renal functions in the fever reactions: In this untoward reaction, the predominant mechanism of renal function-change is homeostatic one which means the constriction of blood vessels in the region of internal organs. But homeostasis itsself do not bring about the abnormal renal circulation. In the patients of hypoproteinemia and dehydrated state, the fever reaction results the various type of Oxford shunting and severe decreasing or renal flow in cooperation of homeostasis and type II of patho-physiological changes in renal function as over montioned.
4. Renal functions in the hemolytic reactions: There are seen the various kinds of Oxford shuntig in every cases of this untoward reactions and the durations of such abnormal renal circulation are more longer. Especially in the severe hemolytic reactions, the abnormal renal functions identify to that of shock reaction as Oxford shunting III.

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The most severe untoward reaction due to blood transfusion is hemolytic reaction which is more often seen in preserved blood transfusion. The cause of this reaction is hemoglobinemia which is resulted by the destruction of red cell. The most important clinical signs of this reaction are the early shock syndrome and late renal failure. To know the patho-phy-siological pattern of this reaction, the author studied on the experimental hemoglobinemia in dogs.
1. In the experimental hemoglobinemia in dogs, the hemoglobin is excreted in urine when the concentration of hemoglobin in blood plasma is over 150 mg/dl. In the group of alkali-urine There is found the parallel relation between the concentrations in blood plasma and in urine. On the contrary, in the group of acid-urine there is no parallel relation in both.
2. The changes in circulatory volumes; In the early stadium of experimental hemoglobinemia, the circulatory plasma volume decreases markedly, especially in the group of acid-urine. The circulatory blood volume decreases in relative smaller grade than that of plasma volume. This means the concentration of blood. The decreasing of circulatory red cell volume is seen.
3. The constrictin of blood vessels in experimental hemoglobinemia: This change was studied by angiography by 70% jod-pyraceton. In the just after stadium there are shown the marked constriction of A. renalis and relative constriction of A. coeliaca or relative dilatation of A. mesenterica. After 30 minutes there are shwon most marked constriction of all abdominal vessels, especially of A. renalis. The over mentioned constriction of vesselsdie to experimental hemoglobinemia identify to the marked sympaticotonic state and may be said as re-distribution mechanism of blood to central organs. The cortical shunting of renal circulation is also shown.
4. Liver function; There are seen relative severe disturbance of liver functions (Takada's reaction, Thymol reaction, CaCl2 reaction etc). The amino-acid curves of blood plasma show the hypofunction of oxygenation in liver, namely anoxia of liver.
5. Renal functions; The renal blood flow and glomerular filtration ratio decrease, but F. F. increases. The non-resorption tatio increases markedly and this means the marked increasing of resorption function.
The author concluded these patho-physiological pattern of experimental hemoglobinemia as the following schema.

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A preliminary study was made on the experimental serum anaphylaxis and the followings were observed. The γ-globulin was decreased by shock, but. the decrease was inhibited by using of intravenous anesthetics (Ravonal) or antihistamic drugs (Anergen). This was quite similar and parallel to the changes of hypotension which was caused by the reinjection of the antigen and inhibited by Ravonal and Anergen.
When the anaphylactic by-effects were experimentally caused by transfusion of the rabbits blood with antigen to the sensitve rabbits, a severe shock resulted and γ-globulin decreased remarkably. By using of Ravonal or Anergen at that time, however, there was seen an inhibition of the decrease of γ-globulin as well as of the hypotention.

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An inadequate heterotypical transfuion in the same kinds was made in rabbits, and the decrease of γ-globulin was less by using Ravonal or Anergen preliminaily, compared with the contrast-group. In the group with Ravonal, the γ-globulin returned almost to the normal limits three hours after transfusion.
The hemolysis in the serum was almost similar in all of the above stated groups 30 minutes after transfusion, but it remarkably decreased in the groups with Ravonal or Anergenafter 3 hours. The hypotension also had the same tendency as γ-globulin and was inhibited by Ravonal and then Anergen.

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Human whole blood was injected to the normal rabbits at the rate of 8-12cc per kg, thus the by-effects were caused.
In the contrast group, a severe shock resulted: some died directly after the transfusion and others died within several hours with anuria or oliguria. In the groups with Ravonal or Anergen, however, these symptoms were inhibited, especially in the group with Ravonal.
The hemolysis was similar in all of these groups, but it decreased in the groups with Anergen or with Ravonal with the lapse of time, especially in the latter.
The γ-globulin increased temporarily in the contrast group and the gronp with Anergen, then decreased markedly with the lapse of time, thus the severe symptom or death resulted. In the groups with Ravonal, however, the γ-globulin slightly decreased temporarily and then rather increased after 3 hours.

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The by-effects of blood transfusion could be clinically prevented by using Ravonal or Anergen before transfusion.
In all cases who had by-effects by transfusion, a marked decrease of γ-globulin was observed.
No distinct changes of the serum protein fractions were seen by the blood transfusion up to the amonnt of 100-125cc, when no by-effects occurred.

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