The author has investigated the influences of a so-called new vitamin B group comprising orotic acid, thioctic acid, pantothenic acid, adenine, and carnitine at the optimal medium concentrations on the outgrowth of bone marrow cells and motility of neutrophils of normal rat bone marrow cultured in vitro, and obtained the following results.
1. Stimulation of the outgrowth of bone marrow cells was seen with carnitine, adenine, pantothenic acid, thioctic acid, and orotic acid in this declining order.
2. Migrating velocity of bone marrow neutrophils was accelerated with pantothenic acid, adenine, thioctic acid, carnitine, and orotic acid in this declining order.

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The author has investigated the influences of a so-called new vitamin B group comprising orotic acid, thioctic acid, bantothenic acid, adenine, and carnitine at the optimal medium concentrations on the number of megakaryocytes and their functions of motility and platelet separation of normal rat bone marrow cultured in vitro, and the results obtained are as follows.
1. The number of megakaryocytes appearing in the outgrowth was increased with adenine, carnitine, pantothenic acid, orotic acid, and thioctic acid in this declining order.
2. Acceleration of the functions of megakaryocytes was observed with carnitine, adenine, pantothenic acid, thioctic acid, and orotic acid, in this declining order.

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A histochemical study on alkaline phosphatase, acid phosphatase, β-esterase, β-glucuronidase, lactic dehydrogenase, glucose-6-phosphate dehydrogenase, succinic dehydrogenase, malic dehydrogenase, isocitric dehydrogenase, glutamic dehydrogenase, α-glycerophosphate dehydrogenase and β-hydroxybutyric dehydrogenase has been carried out to clarify the relationship between the function and morphology of the normal human brain tissues. The materials, 34 in number, were all obtained surgically, and cut at 20μ in -20°C cryostat, then the sections were incubated in each histochemical reaction mixtures. For the histochemical demonstration of hydrolytic enzymes, azo-coupling method was employed, and for the oxidative enzymes, the method described by Pearce with Nitro-BT as the electron acceptor was employed.
The following results were obtained;
1) The characteristics of enzyme distribution concerned with glycolysis, aminoacid and fat-metabolism in cerebral tissues were; a) The enzymatic activities in cerebral cortex were generally stronger than those of the medulla. b) The fluctuation of the enzymatic activities were more marked in nervous cells than in other tissues.
2) Alkaline phosphatase and β-esterase activities were exceedingly low in both cerebral cortex and medulla except for some leptomeninx and capillaries. The activities of aminopeptidase were all negative in the brain tissues.
3) In anerobic glycolysis, lactic dehydrogenase activity was very strong in cerbral cortex, and glucose-6-phorphate dehydrogenase activity was relatively weak.
4) In aerobic glycolysis, the activities of succinic, malic and isocitric dehydrogenases were strong in cortex and weak in medulla. Especially isocitric dehydrogenase activity was exceedingly weak.
5) β-glucuronidase activity was rather higher in medulla, and distributed extremely in the periphery of each cells.
6) In cerebellum, nearly all the enzymatic activities were exceedingly strong in Purkinjes' cells. Especially acid phosphatase activity was the strongest in that cells. The enzymatical distribution in other parts of cerebellum was nearly the same in that of cerebrum.

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The accurate classification of leukemia is very important, because antileukemic agents have different effects for each leukemic types. But still now, it is often difficult, especially in acute leukemias.
The urine hydrolysis test was first proposed by Brachet and then Thoma and Gardner et al, who suggested that nuclear chromatin of myelocytic cells were “hydrolysed”, while lymphocytic and monocytic nuclei remained intact.
The purpose of this study were to establish the most considerable conditions to carry out this test, and to confirm the very responsible substance in urine for this test.
Certainly, the lytic action of the urine on nuclear chromatin were variable in different conditions. Therefore, 3 factors, pH and temperature of the urine and incubation time were discussed for the purpose of standardization of Bracket's test. The temperature of urine specimen was heated at 60°C in preliminary studies. But it was demonstrated that the most effective temperature of this test was 37°C from aspects of its biochemical natures in this paper, so I tried to design the best method. It was the best method with temperature of 37°C with incubation time of 10, 20, 30, and 40 minutes for each blood film, which showed satisfactory results in hydrolyze myelocytic cell nuclear chromatin to differentiate from other cells.
The optimum pH range of the urine for this reaction were different in blood film and in bone-marrow film. Nevertheless, it was noted that the range of pH 6.2 to 6.8 was common and suitable for both films.
The urine substance responsible for this test was studied further. The effect of varying concentration of salt solution and pure distilled water were tried in same method instead of the urine with various conditions but they showed only diffused nonspecific effect, not like the urine specimen. Also purified enzyme solution of RNase in buffered solution and in distilled water resulted nonspecific reaction.
On the other hand, the reaction of crystaline DNase solution applied to blood film were markedly similar to those of the Brachet's test. The DNase activities on the urine, were parallel very well with the results of the Brachet's test. In addition, the activities of the Brachet's test fell down linialy with dilution of the urine by buffered solution. From these, the hydrolysis of nuclear chromatin of cells seemed to be due to DNase of urine most likely.
Especially, DNase I must be responsible for the Brachct's test because the optimum pH and temperature in this test were at pH 6.6 to 6.8 at 37°C, while no activity was seen at pH 5.0. And this test was inhibited by citrate and not by MgSO₄.
In clinical application, it was observed that this Brachet's test was exactly useful in differentiation of the type of leukemic patients.

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Intestinal tumors were studied enzyme-histochemically about the same enzymes as reported in part 1 and about monoamine oxidase. Tumors examined were benign and malignant polyps of large intestine, reticulum cell sarcomas of ileum and cecum, carcinoid of cecum, metastatic carcinomas of small intestine and carcinomas of large intestine.
Alkaline phosphatase was negative in crcinoma but weak activity was sometimes observed in the stromal element. Aminopeptidase was also slightly positive in the stromal element of certain carcinomas and its activity appeared in carcinoma cells occasionally. Generally acid phosphatase, β-esterase and β-glucuronidase reduced their activities in cancer cells but in mucinous carcinoma acid phosphatase was rather strong.
Oxidative enzymes were decreased in carcinoma except lactic and glucose-6-phosphate dehydrogenase, latter two enzymes, on the other hand, usually increased their activities in carcinoma. Oxidative enzymes, however, revealed a strong activity in invading margine and small cell nestle of carcinoma. Monoamine oxidase was stronger in well differenciated carcinoma than in poor differenciated one.
Benign polyp showed similar enzyme activity as normal intestinal epithelium while malignant polyp showed increased oxidative enzyme-activities.
Most enzyme activities were very weak in sarcoma and carcinoid, but glucose-6-phosphate and isocitric dehydrogenase were moderate in the latter.

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In order to pursuit the iron metabolism in the so-called Banti's disease, several investigations about iron metabolism of the erythroblasts in the bone marrow were performed by radioautography and sideroblast examination in the patients with Banti's disease and treated patients with postsplenectomy and administration of iron.
Following results were obtained.
1) In the sideroblast staining of the patients with Banti's disease, the grade of appearence of the iron granules in the erythroblasts revealed considerably lower level than that in the healthy persons. In comparison with normal healthy persons, Fe⁵⁵ uptake into the erythroblasts in the patients with Banti's disease was markedly increased in vitro and to show that Fe⁵⁵ was uptaken by almost all of the erythroblasts and most of cells revealed moderate to markedly high uptake. Above are the similating findings in the essential hypochromic anemia.
2) In the splenectomised cases of 2 months after operation type I of the sideroblasts is increased and this tended to show approaching to the normal. While, radioautographically, Fe⁵⁵ uptake into the erythroblasts clearly showed to be decreased in the same splenectomised case.
In the oral administration of the iron (ferrous gluconate 1 to 2g per day) for 3 months, appearance of type I and II of the sideroblasts was observed to remarkably high grade and Fe⁵⁵ uptake into the erythroblasts showed to be decreased. The erythroblasts that had appeared in high grade tended to show decreasing in number. In the intravenous administration of iron preparate (dextran Fe 100mg/500ml/day) for one week, there were increasing sideroblasts and markedly decreasing Fe⁵⁵ uptake into erythroblasts and these tended to show similating finding in the aplastic anemia.
3) As noted already in the various blood diseases, generally, a negative correlation was observed between average number of stainable iron granules and average rate of radioactive iron appearance in the erythroblasts before and after treatments of the patients with Banti's disease; appearance of radioactive iron was observed in high grade in the decreasing condition of the stainable iron granules and also there is a tendency to reveal the reverse findings mentioned above. It is presumed that quantity of non-hemin iron plays an important role in the iron uptake of the erythroblasts.

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In order to persuit the iron metabolism in Banti's disease, distribution and fractionation of the radioactive iron (Fe⁵⁹) and non-radioactive iron in the red cells, serum and various organs such as liver or spleen were investigated by intravenous and oral administration of Fe⁵⁹ to the rabbits given serum or extract of the spleen of the patients with Banti's disease and the rabbits sympathectomized of the splenic branch.
Following results were obtained.
1) On the intravenous administration of Fe⁵⁹ to the rabbits given patient serum of Banti's disease, the radioactive and non-radioactive iron tended to show increased distibution in the liver and spleen. It is clear from this finding that preservation of the iron in these organs is taken place.
2) In the rabbits given extracts of the spleen of the patients with Banti's disease, there are differences of the iron distribution between intravenous and oral administration of Fe⁵⁹; in the former almost the same results as in the serum administration was observed, while in the latter it is shown no remarkable changes in the organ distribution or rather somewhat decreasing tendency. These findings are suggestive of a possibility of the absorption disturbance of the iron in the gastrointestinal tract.
3) On the intravenous administration of Fe⁵⁹ to the rabbits sympathectomized of the splenic branch, the labeled isotope tended to show increasing in the liver and spleen as seen in the serum administrating animals. This finding apparently indicated that preservation of the iron to the organs is occurred.
4) It is reasonable to presume from above findings that the preservation of iron to the organs such as liver or spleen plays an important role for anemia and iron deficiency condition of the erythroblasts in Banti's disease and also it is suggested that a possibility of disturbance of iron absorption could be existed in the gastrintestinal tract.

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Recently, having encountered a case of multiple myeloma, we have described about her X-ray findings, results of clinical examination and given our comments on differentiated findings with these of other diseases appearing in the literature referred to.

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Recently, as we have found two cases (42 years old male, 54 years old male) of gastric cancer resulting from stomach ulcer, we have described about these X-ray findings and reported the differentiated aspects with those of other diseases appearing in the literature referred to.

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This work was undertaken to examine dynamic aspects between fibrinolytic factors and clinical findings in chronic hepatitis, precirrhosis and cirrhosis of the liver.
The results were as follows:
1) Increasing activity of plasma clot lysis (PCL) and euglobulin clot lysis (ECL) were seen in precirrhosis and cirrhosis of the liver.
Total fibrinolytic activity (Eug+SK) decreased in cirrhosis of the liver.
2) Increase or decrease in activity of inhibitor and activcator was not seen on a certain type of the chronic liver diseases.
3) PCL activity was closely correlated to the degree of BSP retention test, and also increasing antiplasmin activity was parallel to PCL activity.
On the other hand, positive serum colloidal reactions were more closely related to PCL activity than BSP retention test showed. moreover, no clear cut relationship between serum transaminase (S GOT, S GPT) and fibrinolytic factors was not recognized.
4) With regard to hemorrhagic tendency in chronic liver diseases, most cases which presented increased activities of PCL and ECL, have prolonged bleeding time and positive Rumpel-Leede phenomenon. On the contrary, reverse relationship between both phenomena above descrived was not obtained. The cases presented positive thrombotest had hemorrhagic tendency, and also the cases with hemorragic tendency showed positive thrombotest.
5) There was no relationship between morphological changes of the liver and fibrinolytic phenomena in chronic disases of the liver. but most cases with the damage of limiting plate in Glisson's capsle of the liver presented increasing activity in PCL.
6) In the relationship between a period of time of the illness and fibrinolytic factors, fibrinolytic activities have been increaed in PCL and ECL.
7) In considering fibrimolytic phenomena, it must be emphsized that it was very much important to detect varying fibrinolytic factor and must be considered a clinical significans with dynamic homeostasis of the fibrinolytic factors.
8) Most cases had no remarkable disorder of a homeostasis of fibrinolytic phenomenon on disease of the liver, eapecially even in both precirrhosis and cirrhosis of the liver.

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Clinical obsavations on fibrinolytic phenomena have been presented in chronic diseases of the liver (precirrhosis and cirrhosis of the liver, chronic infectious hepatitis and lupoid hepatitis) and the author examined activities of fibrinolytic factors in the course of the diseases above mentioned.
The results were as follow;
1) It is necessary to examine repeatedly an activity in fibrinolytic factors at a short interval of time in a course of each disease of the liver.
2) Even if fibrinolytic activity showed to bo in normal range, the disorder of homeostasis in this systm was possible to find by detecting several fibrinolytic factors.
3) Except when showed severe disorder of fibrinolytic phenomenon, reasonable observations in this system were possible by several determining methods of the fibrinolytic factors.
4) It is better to evaluate serum fibrinolytic activities in clinical cases by the determinations of various kind of fibrinolytic factors (whole plasmin, activator and inhibitor, etc).
5) Appearance of hemorrhagic tendency in these cases was followed by preceding latent disorders of fibrinolytic phenomenon.
6) In general, the cases being administrated cortcosteroids drugs showed transiently increasing activity of fibrinolysis, but this increase in activity recovered reversibly to normal value after a short period of time.
7) The direct correlation between fibrinolytic activity and transaminase activity in serum was not recognized.
8) In the cases with increase in serum transaminase activity after administration of corticosteroids drugs latent activations of fibrinolytic activator were presented, thereafter these cases showed to appearent activation of fibrinolytic phenomenon.

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The Mycobacteria contain large amounts of lipids and several of these lipids are biologically active. Fatty acids that are utilized for lipid synthesis are synthesized from 2C-unit as acetate or malonate through the malonyl CoA pathway.
In the present report, the quantitative and qualitative changes of activity for the fatty acid synthesis were studied at the varying age of culture. The studies on the activities of the cell-free extracts of H37Ra strain cultured for 2, 3, 4 and 5 weeks for the incorporation of acetate or malonate carbon into long chain fatty acids were carried out.
1. In the case of either acetate or malonate for substrate, the highest activity was observed in the cell-free extract taken from the Mycobacteria cultured for 2 weeks. The activity was reduced for farther duration of culture over 2 weeks.
2. The distribution of radioactivity incorporated into long chain fatty acids from either acetate-1-¹⁴C or malonate-2-¹⁴C was checked with gas liquid chromatography. At any age of culture, the incorporation into hexacosanoic acid was the highest. And the incorporation rate into hexacosanoic acid and fatty acids of fewer than 26 carbons was decreased with prolonged culture, but the incorporation rate into fatty acids of more than 26 carbons was increased with the prolongation of culture age.

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By means of bone marrow tissue culture method deviced by Hiraki, the author investigated the effects cytochrome C on the thrombopoietic function in bone marrow and obtained the following results.
1) Cytochrome C diluted in several concentrations was added to the bone marrow tissue culture of normal guinea pigs, normal persons and patients with various diseases. Average number of megakaryocytes in the growth zone and its thrombopoietic function were examined.
As a result, it has been seen that the thrombopoietic functions of normal bone marrow added with an optimal amount of cytochrome C were moderately accelerated both in guinea pigs and human beings as compared with that of control added with saline.
2) The cytochrome C and each of three kinds of carcinostatic substances; mitomycin C, trenimon, and carzinophyllin were added to the bone marrow tissue culture of normal persons the effects of these drugs on the thrombopoietic functions in the bone marrow were investigated. On the other hand, the bone marrow of guinea pigs of which bone marrow had been rendered to hypoplastic by repeated administration of the above described carcinostatic substances and then treated with repeated administration of cytochrome C were cultured. The thrombopoietic functions of megakaryocytes in these bone marrow tissue culture treated with cytochrome C were maintained moderately high as compared with the control treated with carcinostatic substances and saline only.
From these results it could be concluded that cytochrome C has the accelerating effects on the thrombopoietic function of normal bone marrow as well as the protecting effects on the thrombopoiesis from the myelodegenerative especially thrombopoiesisinhibiting action of several carcinostatic substances.

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By means of bone marrow tissue culture the author investigated the effects of cytochrome C on leukopoietic function and obtained the following results:
1) When cytochrome C was added directly to the bone marrow tissue culture of normal guinea pigs, normal persons and patients with various diseases the growthrate of outgrowth, the wandering velocity and carbon particle phagocytosis of neutrophils were increased as compared with those of the control added with saline.
As the result, it has been seen that by adding an optimal amount of cytochrome C the leukopoietic functions of bone marrow were accelerated.
2) When cytochrome C was added with carcinostatic substances to the bone marrow tissue culture of normal persons, the growth rate of outgrowth, the wandering velocity and the carbon-particle phagocytosis of neutrophils were increased as compared with those of the control added with only carcinostatic substances. The similar results were obtained in the case of bone marrow tissue culture of guinea pigs of which bone marrow had been made hypoplastic by repeated administration of carcinostatic substauces and then injected with the repeated administration of cytochrome C.
On the other hand, in its clinical applications, the repeated administration of cytochrome C were effective on some cases of leukopenia induced with carcinostatic substances.
As it has been demonstrated that cytochrome C has some relieviug effects on the disturbance of leukopoietic functions of bone marrow successively injected with carcinostatic substances.

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The present communication deals with the study on an attempted suicide case who had taken a large dose of an agricultural drug, EPN, but was successfully saved. Some comments have also been given about her clinical findings with reference to the reports available on similar cases.

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Of 180 cases with acute and chronic hepatitis and liver cirrhosis treated with glucocorticoids, 33 cases of steroid diabetets were selected for this study. Atmpts were done on the relation of a mode of occurrence and the clinical feature of steroid diabetes to pathologic changes of the liver.
The results were as follows:
1) Steroid diabetes was found in seven cases of 39 acute hepatitis (17.9%), nine of 65 chronic hepatitis (13.0%) and 17 of 72 liver cirrhosis.
2) The incidence of steroid diabetes was slightly high in the obese cases and patients with serum hepatitis, and significantly high in patients with diabetic desposition.
3) There was no significant difference among Prednisolone, Dexamethasone and Paramethasone in the incidence of steroid diabetes. Although the cases treated with both Norandrostenolone phenylpropionate and Androstanolone showed no differece from those with glucocorticoid alone, the cases treated with the combination of Oxymetholone showed extremely high incidence of steroid diabetes. (eight of 17 cases; 47.1%)
4) Steroid diabetes could be classified into two types; early developed and lately developed type, according to the time of developmnt and the clinical feature The former showed clinical findings of diabetes. On the contrally, no diabetic findings were shown in the latter.
5) In liver cirrhosis, more early developed types of steroid diabetes were fouud and in acute and chronic hepatitis, no difference was seen in the incidence of the two types.
6) In the early developed type, a considerable number of cases showed heredity of diabetes, obesity, fatty degeneation of the liver cells and vacuole degeneration of nuclei at a punch biopsy, and abnormal glucose tolerance test (not diabetic) before the treatment of glucocorticoids. On the other hand, the lately developed type did not show those above mentioned.
7) Diabetic desposition or latent diabetes and abnormal glucose metabolism caused by a original disease were found in many of the early developed type, and few in the lately developed tvpe. It is presumed that these differences played a important role in the time of the development and the clinical feature of steroid diabetes.
8) The incidence and the clinical feature of steroid diabetes, and the time of onset and disappearance of glycosuria were not always closely related to the degree of the disturbance of liverfunctions. However, they were closely related to the fatty degeneration of the liver cells and vacuole degeneration of the nuclei.
9) In many cases of steroid diabetes, the fatty degeneration of the liver cells and vacuole degeneration of the nuclei appeared or progressed after the glucocorticoids treatment. However, no tendency was seen in distribution of fat in acini of the liver.

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A comparative study was undertaken to examinie the relationship between the cases with steroid diabetes developed after treating with glucocorticoids and the cases not developed after the same treatment, by means of various loading tests including glucose, insulin, prednisolone-glucose, tolbutamide and compound multiple amino acids performed before and after the treatment, The cases studied included of acute and chronic hepatitis and liver cirrhosis. The results obtained are as follows.
1) Nine out of 14 case with the early developed type of steroid diabetes showed glucose assimilation index (KG) of diabetes mellitus before the treatment with glucocorticoids. Five out of 7 cases with the lately developed type showed KG indicated borderline value between normal healthy person and diabetes mellitus. On the other hand, abnormal KG was also seen in 19 out of 29 cases not developed steroid diabetes From these results it was found that there was no significant correlation between KG and development of steroid diabetes.
2) KG after glucocorticoid administration decreased in many of cases with the early developed type of steroid diabetes. On the other hand, KG in cases with the lately developed and not developed showed no significant change between before and after glucocorticoid administration.
3) Insulin sensitivity tended to decrease after glucocorticoid adminstration, but it was no significant difference as compared with that before glucocorticoid administration. Insulin sensitivity after glucocorticoid administration in cases with steroid diabetes developed tended to decrease as compared with that in cases not developed.
4) There was not always close relation between the influence of prednisolone on intravenous glucose tolerance test and the appearance of steroid diabetes.
5) Some of cases with the early developed type showed diabetic response in tolbutamide tolerance test, but no diabetic response was found in cases with the lately developed and not developed type.
6) The rate of increase in blood sugure, pyruvic acid and amino-N in blood and urine after giving glucocorticoid Its rate was found to be higher in cases with steroid disbetes deveoloped either early or lately as compared with cases not developed, showing the similar rate to that after administration of ths compound multiple amino acid in diabetes mellitus.
7) The rate of increase in blood sugure after administration of the compound multiple amino acids tended to be higher in cases treated with the combination of glucocorticoid and anabolic steroid as compared with cases treated with glucocorticoid alone.
8) There was no significant correlation between the results of various loading tests including glucose, insulin, prednisolone-glucose, tolbutamide and compound multiple amino acids and the findings of biochemical liver function tests and liver biopsy.

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