Virus preparations were obtained from mammary tumors which developed spontaneously in C3H mice by three different methods; 1) filtration through Berkefeld-N or Chamberland L3 filters, 2) differential centrifugations, and 3) fluorocarbou extraction. Each virus preparation was used for immunizing rabbits, and was then evaluated for the degree of purification of the viral antigens by direct immunofluorescent staining. Following results were obtained: 1. The fluorocarbon extractin method was more excellent for the purification of mammary cancer virus than filtration or differential centrifugation method. 2. Staining procedure of the direct immunofluorescence was improved as follows; frozen sections were fixed with cold acetone-fluorocarbon mixture instead of coid acetone, and duration of staining was prolongel over six hours in a cold moist chamber. As a result it was possible to observe clearer and more numerous fluorescence than by the direct staining of Coons' original procedure.
As mentioned in the author's previous study, the complement method of immunofluorescence allowed to observe clearer fluorescence of the viral antigen than by the direct and indirect method. In this study, therefore, localizaton of viral antigens of C3H mammary cancer was persued with the fluorescescent serum that reacted immunologically with the complement. 1. The viral antigen was observed in tumor cells and intercellular spaces and particularly rich in perivascular portions. 2. In the metastastatic lymph nodes viral antigens were observed in some tumor cells, and in the spleen of tumorbearing mice they were observed within retculum cells on rare ocasions. 3. Intracellular localization of viral antigens was spread in the peripheral portions of the cytoplasm of tumor cells, and no specific fluorescence was observed in the nucleus of tumor cells. 4. No specific fluorescence was seen in the liver, kidney, lung, bone marrow and brain of the tumor-bearing C3H mice and in tissues of normal mice. 5. In mice with spontaneous mammary cancer, viral antigens were encounted more frequently than in mice with transplanted tumors.
During serial inoculation of spontaneous Iympbocytic leukemia in C58 and AKR mouse strains by splenic cell-free extracts, a change of the leukemia type from lymphocytic to myelogenous was observed. As a method to clarify this phenomenon, isolation of viruses by passaging through guinea pig brains and cross-neutralization tests of viruses according to the method of Hamazaki were performed with these two types of leukemias in both strains of mice. First of all, the author succeeded in isolating viruses of the, original lymphocytic leukemia and the changed myelogenous leukemia. These viruses induced a mesenchymal tissue reaction in the spleen, lungs and liver of the guinea pigs in serial passage. The pathological changes were especially prominent in the lungs and spleen; in the lungs so-called viral interalveolar pneumonia and granulomatous inflammation around the interstitium were observed, and in the spleen proliferation of the reticulum cells in the red pulp and of the sinus endothelium were seen. These chauges were slight in the liver and brain. A diffuse proliferation of the stellate cells in the liver and chickening of the pia mater and sometimes proliferation of the glia cells were observed.
In the previous experiment, the author succeeded in isolating viruses of the original lymphocytic leukemia and the changed myclogenous leukemia in C58 and AKR mice. In the present experiment, the author performed cross neutralization tests between these viruses according to the method of Hamazaki. Though coss neutralization in C58 leukemias were generally observed, strictly speaking, antigenicity of the original and variant viruses were not completely identical. The author obtained the same results with the AKR leukemias. The viruses of the original lymphocytic leukemia and the changed myelogenous leukemia were found to be immunologically closely related and a change of the leukemia type was considered to be due to the difference in the tissue affinity of these two different viruses originally co existed.
In the second report, by the method of cross neutralization tests, the author investigated seroimmunologically the possible correlation between the viruses of the original lymphocytic leukemia and the changed myelogenous leukemia in both strains of mice. In this experiment, with the purpose of further clarifying the immunological correlation among four viruses, the author performed cross neutralization tests of AKR mouse leukemia viruses with antiserum against C58 mouse leukemia viruses. As a result, cross neusralization between the viruses from C58 and AKR mice did not occur and the leukemia viruses from these two strains were shown immunologically to be considerably different. From all these facts reported in the present series of papers and other immunological experiments in our department, a change of the leukemia type was considered to be due to the difference in the tissue affinity of these two different viruses originally coexisted.
Alterations of serum lactic dehydrogenase (S-LDH) activity and isozyme during therapy in tumor-bearing mice were assayed. 1. Normal Strong A mice receiving Mitomycin C (MMC), Chromomycin A3 (TM), and Chloroquine phosphate (CQ) respectively were evaluated for S-LDH activity. S-LDH activities of the mice receiving MMC and TM were about twice as high as mormal, this seems to be the side effects of chemotherapeutic agents. The strong A mice receiving CQ showed no changes in S-LDH activity. 2. The strong A mice bearing Bashford carcinoma and the Swiss mice bearing fibrosarcoma were evaluated at weekly intervals for S LDH activity and isozymes. S-LDH activities gradually elevated for 4 weeks. In the Strong A mice bearing Bashford carcinoma LD3 increased and LD1 decreased gradually. In the Swiss mice bearing fibrosarcoma LD3 was higher than normal at every week. 3. S LDH activities in tumor-bearing mice undergoring therapy with MMC and TM, especially MMC, markedly decreased correlating with tumor regression. As to the S-LDH isozyme, LD3 in mice receiving TM decreased correlating with decrease of S-LDH activity. In mice receiving others no characteristics were observed. 4. In couclusion, the assay of S-LDH activity and isozymes during therapy directing at tumors are useful in determining wheather or not a chemotherapentic agent affects the tumors.
The effects of antigenic and non-antigenic stimulations on the cell output through the efferent lymph from the popliteal lymph nodes of adult rabbits were studied. The chief results obtained are as follows: 1. The cell output through the efferent lymph flow is greatly increased by antigenic stimulations and the magnitute of increase in the cell output runs almost parallel to that of the increase in nodal weight. 2. Heterograft causes a marked increase in the cell output from regional lymph node, whereas homograft hardly produces such an effect. 3. Complete Freund's adjuvant with Mycobacteria also increases the cell output from lymph node to a considerable extent, whereas incomplete Freund's adjuvant without Mycobacteria is less effective. 4. However, when injected with bovine gamma-globulin, incomplete Freund's adjuvant also produces a marked increase in the cell output from lymph node. 5. The increase in the cell output from lymph node produced by antigenic stimulation is defenitely accompanied by the appearance of large pyroninophilic blast cells in the efferent lymph. In contrast, complete Freund's adjuvant or incomplete Freund's adjuvant alone is not able to elicit an appearance of these cell in the efferent lymph. 6. An incresse in cell output through the efferent lymph from the regional lymph node, and the occurrence of large pyroninophilic cells in the efferent lymph, particularly the latter, are considered to be specific to antigenic stimulation. Therefore, these reactions may be used as measures to test the antigenecity of stimulating agents.
Mycotic and clinical studies were carried out on gastric ulcers with fungi in their bottom, For this purpose 136 stomachs with ulcer were collected by means of operation. The results were as follows: 1. The gastric ulcers with fungi in their bottom were found in 38 cases out of 136 (27.9%). 2. Massive bleeding showed 13 cases out of these 38 (34.2%). 3. As for the majority of the gastric ulcers with fungi in their bottom clinical manifestations were as follows: in most cases, the duration of complaints was more than 1 year, the size of the ulcers was more than 1cm, the depth of the ulcers reached to the muscle layer, the site of the ulcers was the angle or the body of the stomachs, the patients were middleaged or old aged and the sex of the patients was male. According to these clinical findings the gastric ulcers with fungi in their bottom were regarded as a chronic and a hard to be cured type. 4. Most of the fungi found on the surface of gastric ulcers were identified as Candida albicans.
Studies were performed on the relationship between hard cured gastric ulcers and fungi in rats. Experimental gastric ulcers were prepared by the use of the Clamping-Cortisone method. Rats with experimental gastric ulcers were divided into 3 groups, A, B and C. Group A was neither administered drugs nor fungi. Group B was given Candida albicans. Group C was dosed chlorhexidine. The results were as follows: 1. Experimental gastric ulcers with fungi in their bottom were found in 4 rats out of 13 belonging to group B. 2. The delection of fungi on the crate surface of the experimental gastric ulcers revealed that positive results were obtained in all cases of group A or B and in 3 cases out of 11 of group C. As to the species of the fungi, in group a Torulopsis famata, in group B Candida albicans and in group C. Torulopsis famata were identified respectively. 3. The normal fungous flora in the intact stomachs of experimental animals was identified as Torulopsis famata. 4. The fungous form in the bottom of experimental gastric ulcers was yeastlike in the cases that were sacrificed 3 weeks later making ulcers. In case the heading of experimental gastric ulcers was protracted, there were seen mycerial fungi increasing in quantity. 5. The crater surface of the gastric ulcers of group B where fungi grew abundantly was dirty histlogically and the recovery of gastric ulcers was delayed. The scar formation of gastric ulcers of group B was weaker than group A and C. These results apparently indicate that the course of the gastric ulcers with fungi in their bottom will be delayed in group B. 6. The recovery of experimental gastric ulcers in group C was most remarkable among 3 groups. It may be posible that the application of chlorhexidine as the gastric ulcer agent will contribute to the purification of the surface of gastric ulcers and promote the healing of them.
1. The rabbits of a commerical stock were immunized by the technique of thermostable antigen with Candida albicans of IFO 0583 strain. The agglutination titer of the immune serum was 1: 5120 against the hemologous organism. 2. The antiserum of rabbits was labeled with the fluolescein isothiocyanate. The agglutination titer of the labeled antisterum was 1: 160 against Candidd albicans (the homologous organism) as well as Torulopsis famata of normal flora in the rat-stomach. The common determinants of the two strains were also identified by the precipitation reaction. 3. The mono specific antiserum was obtained by absorbing the above mentioned labeled serum with Torulopsis famata. The thus absorbed labeled antiserum showed the agglutination-titer 1:5 agcipitate in the Ouchterlony method was found against only Canaida albicans. The gamma globulin concentration of the fluorescein isothiocyanate-labeled, absorbed and monospecific antiserum solution was 0.9mg/ml. 4. Fungi in the gastric ulcers of human and rats could be identified as Candida albicdns by its specific fluorescence when stained with the fluorescein isothiocyanate-labeled, absorbed and mono-specific antiserum.
The occurrence of serum hepatitis in the Okayama University Medical School and its affiliated hospitals for a period from September 1966 to June 1967 was studied. A special attention was paid on the usefulness of volantary blood donor for the pervention of serum hepatitis. The results were as follows: 1. Out of 178 cases, 18 (10.1%) developed serum hepatitis, and 3 (1.7%) were jaundiced. The incidence of serum hepatitis in the subjects who recieved blood from voluntary donors was apparently lower than that of the subjects received blood from paid donors. 2. Among the causative etiology for serum hepatitis, the area of donors residence was proved to be as important as the amount of transfused blood. 3. A histry of viral hepatitis was found at the rate of 0.8% and the elevated activity of S-GOT over 40 units was observed at 0.7% in the blood donors. The donors with a previous history of viral hepatitis and elevated activity of S GOT in the same subject was rather rare, and it was scarcely found overlap of the history and abnormal S GOT. 4. It was re-confirmed that the examination of S-GOT of donors was valuable for the screening of infective donors on the basis of age distribution of the donors with abnormal S-GOT as well as reduced incidence of serum hepatitis through exclusion of the abnormal donors. 5. The residence areas of the infective donors were in accordance with the areas where infective hepatitis has been preveiled.
Of 137 blood recipients from the blood donors with a history of infectious hepatitis, 61 recipients were examined in terms of liver function tests for more than 3 months. The results obtained were as follows: 1. There was no incidence of serum hepatitis except 2 cases in the recipients who received blood from the donors who had been suffered from hepatitis before 1956. 2. Out of 29 cases, 9 cases including 4 cases icteric serum hepatitis were proved to be the subjects received blood from the donors with a history of hepatitis after 1957. Comparing with the incidence rate of serum hepatitis by the blood transfusion from the voluntary donors for a same period. The above mentioned rate is notably high, and this is almost equal to the rate of the incidence in the cases received blood from paid donors. 3. The incubation prid was within 30 days in 7 out of 11 cases with serum hepatitis. 4. The mixture of blood from the subjects with a history of viral hepatitis was found to be only one or two units, therefore the incidence of serum hepatitis was not increased with the amount of transfused blood. 5. The residential areas of the donors were in close association with the epidemic areas of infectious hepatitis. 6. Consequetly, it is concluded that those donors who have a history of hepatitis for the past 10 years and who have residence in the epidemic areas, should be eliminated from the choice of the blood donors.
HIM test was performed on the 103 patients with liver diseases and 33 cases of the other diseases. The results obtained were as follows; 1. The titer of HIM antibody showed high percentage in the sera of the patients with viral hepatitis. The titer of HIM antibody in serum hepatitis tended to be higher than that of infectious hepatitis. 2. In acute viral hepatitis the titer of HIM antibody was usually elevated at the beginning of the diseases, then the titer decreased gradually when the time lapsed. None of the cases of viral hepatitis were observed, in whom the HIM test was negative at the beginning and it became positive in the course of the diseases. 3. The family of the patients with infectious hepatitis tended to show positive results in the test. 4. The suppressive effect on the producton of HIM antibody was not able to prove by the administration of immunosuppressive drugs such as corticosteroids, 6-MP and Imuran. 5. The positive results were observed in some cases of healthy population, leucemia, Hodgikin's disease and chronic nephritis. It was however, difficult to assume that the antibody observed in the sera of viral hepatitis is consistent with the antibody proved in the sera of the other diseases.
The specific gravity of blood was studied with special reference to dietary habitus of the subject. The materials studied were 46 student nurses and 73 housewives. The results were as follows; 1. The specific gravity of blood was in close relation to the quantity and hemoglobin content of the red corpuscles. The protein content of the plasma was proved to be another major factor of the specific gravity. 2. In the group of disqualified subjects, below 1.052 in the specific gravity, the following findings were obtained; (1) Low level of serm iron and dietary iron intake were apparent. (2) Hypoacidity or anacidity was usually present. (3) Daily labor hours and menstrual period had a close relation to the specific gravity of blood. 3. The specific gravity of blood showed seasonable changes. 4. The most effective treatment of the disqualified subjects was the improvement of dietary habits.
This paper is to report a case of nasopharyngeal tumor. The patient was a 57 years old man admitted to this hospital with chief complaints of impaired hearing and headache. The diagnosis had not been made until the biopsy of the enlarged lymphnode in the right neck was performed and histologically examined. The patient died in spits of radiation therapy with 60Co over the neck. Postmortem examination revealed undifferentiated cell carcinoma in sphenoethomoidal sinus and its metastasis to the liver, lumbar vertebrae, and lymphnodes of various area. Studies on 41 cases of nasopharyngeal tun or experienced in the department of Otorhinopharyngol-aryngology of this University in recent 10 years (1957-1966) showed that a large number of the patient as much as 24% of the cases, first visited the clinic of internal medicine of examination of enlarged lymphnodes in the neck or headache.
A 29 year old male who had been transfused whole blood from apparently healthy 6 donors supplied a 40 year old female his blood 14 days before he developed serum hepatitis. The female recipient developed serum hepatitis at 30th day since the blood transfusion. These two cases suggest that viremia and its infectivity occur before the patient exhibit a functional impairment of the liver.
The effects of fasting on the cell output through the efferent lymph from the popliteal lymph nodes of adult rabbits were studied along with the nodal changes. The most striking effects of acute fasting is a marked decrease in the cell output through the efferent lymph. This is chiefly due to a reduction in the efferent lymph flow. The cellular content of the efferent lymph remains almost unchaged, although the nodal weight is much reduced by fasting. The reduction in the nodal weight, on the other hand, runs almost paralled to the decrease in the cell output through the efferent lymph. After stimulating the node by the subcutaneous injection of sheep blood into the foot-pad, simulatneously with the initiation of fasting, the cell output through the efferent lymph is increased to a great extent. This is due to the increase not only in the cellular content of efferent lymph but also in the efferent lymph flow. The weight of the node is also much increased as compared with that of the opposite side. The nodal weight rises almost parallel with the increase in cell output, so that the cell output per unit weight of nodal tissue remains almost unchanged even after the stimulation with antigen. The cellular content of the efferent lymph consists almost exclusively of small lymphocytes in normal and fasted rabbits. After the stimulation with antigen, larger pyroninophilic lymphocytes, especially huge immature blast cells (lympogonia or immunoblasts) defenitely appear in the efferent lymph in large numbers even in the fasted rabbits. Based upon these findings, the effects of fasting on the immune cellular proliferation are discussed.
Clinicopathological and experimental studies were performed to clarify the genesis of primary lipoid pneumonia. Complete examination of resected lungs from six cases, which were believed to be suitable for this entity, were made. In addition to characteristic pathologic changes such as massive appearance of intralveolar foam cells, peribronchial and perivascular inflammation and interstitial fibrosis, definite changes of pulmonary arteries and veins of various size were noted. Pathologic changes in vessels were most prominent especially in the arteries of small and medium size. Definite narrowing of the lumen due to swelling of intima and thickening and swelling of both media and adventitia were noted. These changes were minimum in the large arteries. Occlusion due to thrombus was noted only in one case of medium sized artery. Based on these findings, experimental studies were undertaken to produce the similar pathologic changes in the lungs of dogs and rabbits. Experimental animals were divided into eight groups, where different experimental procedures were applied as follows. Ligation, to cause either occlusion or stenosis of pulmonary artery, vein or bronchus, was performed in five groups of dogs. Vasopression was injected daily for 30 successive days with the dose of 0.5cc. per kilogram of body weight in another group of rabbits. Intrapulmonary inoculation of Staphylococcus aureus was made with 0.3cc. of the bacterial suspension, containing 10mg. of the bacteria in 1cc. of physiologic saline, to induce pulmonary infection, directly through the thoracic wall. The other group of rabbit was fed daily with chow containing 1gm. of cholesterol for 30 days. Experimental animals were sacrificed after a given interval and the lungs were examined for both macroscopical and microscopical pathology. Among the eight groups, the similar pathologic findings to clinical cases, particularly to those in pulmonary vessels, were found only in the lungs of those dogs, whose pulmonary arteries were ligated. Tissue anoxia due to disturbance of pulmonary circulation was considered to be important causal factor for development of primary lipoid pneumonia.
In acatalasemia, specific oral gangrene called Takahara's desease is often encountered. With the purpose to elucidate the causative factor of this gangrene, animal experiments were carried out. Namely, ducks having only a trace amount of catalase in their blood were used as experimental animals and for the control, rabbits were served for the experiments. By injecting various. concentration of hydrogen peroxide in isotonic saline under the mucous membrane of the palate in each animal, macroscopical investigation as well as histochemical investigation of activity of succinic dehydrogenase system were conducted at a fixed time after the injection. The results obtained were as follows. 1) Macroscopically in the ducks there appeared redness, swelling, coat and gangrene resembling the one observable in acatalasemia cases, but in the rabbits redness and swelling were slightly observed. 2) The histochemical findings observed in experiments in the ducks showed marked decrease in activity of succinic dehydrogenase system in mucous epithel, blood vessel wall and muscle compared to that observed in rabbits. It is assumed that the facts described above proves that the succinic dehydrogenase system possessing a strong affinity to hydrogen peroxide are affected in ducks more strongly by the oxidative action of hydrogen peroxide than in the case of rabbits.
With the purpose to know how is the effect of hydrogen peroxide on the succinic dehydrogenase system, experimental studies were conducted using duck, rabbit and mouse. The results obtained were as follows. I. In vivo study After injection of hydrogen peroxide in isotonic saline into the palate of ducks, having only a trace amount of catalase in their blood, inhibition of the activity of succinic dehydrogenase system was observed in the homogenate of their palate. Activity of succinic dehydrogenase system in the homogenate of palate of ducks, injected four times, decreased more markedly than that of duck injected ones. II. In vitro Study 1) Activity of succinic dehydrogenase system in the homogenate of duck palate or mouse muscle, containing less amount of catalase, or in the mouse liver homogenate with addition of NaN3, decreased more markedly than that of rabbit palate or mouse liver containing more amount of catalase. 2) Activity of succinic dehydrogenase system in the homogenate of the various tissues was decreased by addition of hydrogen peroxide. On the other hand, inhibited activity of succinic dehydrogenase system by hydrogen peroxide recovered as time lapses. From the above findings it is considered that the activity of succinic dehydrogenase system inhibited by hydrogen peroxide is reversible. Therefore, it seems that the causative factor of the gangrene in acatalasemia cases lies in the fact that so called respiratory enzyme group such as succinic dehydrogenase system in tissue cells, are disturbed by the accumulation of hydrogen peroxide produced from hydrogen peroxide generating micro organisms (hemolytic streptococcus, pneumococcus etc.)
I. Serum Electrolytes and Blood Glucose in the Development of Gastric Ulcer Following Adrenalectomy Bilateral adrenalectomy in rats caused increase in potassium and decrease in sodium in serum and also decrease in blood glucose level, and resulted in development of ulcer on the gastric mucous membrane. When the above unbalance was corrected with either the oral administration of 1% sodium chloride solution or intramuscular administration of cortisone (0.05 to 0.1 mg., per 100 gm. of body weight, per day), the development of the ulcer was prevented. However, administration of higher dose of cortisone alone (5 mg. per day, per rat) did not prevent the ulcer formation, resulting in hypernatremia, hypopotassemia and hyperglycemia. The prevention was obtained by additional oral administration of 1% potassium chloride solution. In summary, these results are highly indicative of the presence of some direct or indirect relationship between the unbalanced serum electrolytes and blood glucose level due to adrenal deficiency and the deterioration in the local resistance of gastric mucosa against the digestive process to develop subsequent ulceration.
I. Serum Elecurolytes and Blood Glucose in the Development of Gastric Ulcer Following Hypophyseoadrenalectomy Combining the adrenalectomy with the hypophysectomy, the ulceration of gastric mucosa was also observed, accompanied by hyponatremia, hyperpotassemia and hypoglycemia. Making correction of this abnormalities with intramuscular application of cortisone (0.1 mg., per 100 gm. of body weight, per day) simultaneously with oral intake of 5% glucose solution, the ulceration was remakably inhibited, whereas administration of 1% saline alone failed to correct the electrolyte unbalance and to inhibit subsequent ulcer formation. II. Influence of Vagotomy in the Development of Gastric Ulcer Following Adrenalectomy and Hypophysectomy Bilateral vagotomy supressed the causation of the previously mentioned abnormalities in the serum electrolytes and the fluctuation of blood sugar level and also surpressed the development of gastric ulcer, following the adrenalectomy and hypophyseoadrenalectomy. The effect of the right vagotomy on this supression was predominant over the left vagotomy. In summary, these results are highly indicative of the presence of some direct or indirect relationship between the unbalanced serum electrolytes and blcod glucose level due to àdrenal deficiency and the deterioration in the lccal resistance of gastric mucosa against the digestive process to develop subsequent ulceration. Association of vagal nerve in the mechanism of this type of gastric ulcerogenesis was also demonstrated.
Experimental and clinical study on blood coagulation during profound hypothermia either by surface cooling or by extracorporeal circulation was made and the effect of hemodilution in the extracorporeal circulation was investigated. The result was summarized as follows; 1) Profound hypothermia by surface cooling caused decrease in the platelet count and reduction of its function. The count recovered to normal level soon after rewarming but the functional insufficiency remained longer even after the rewarming. The profound hypothermia for less than one hour gave no significant influence on clotting factors other than the platelet. 2) After the extracorporeal circulation alone, decrease in platelet count and the damage on clotting factors were generally observed, but they were temporary and recovered within 24 hours in nearly all cases. However, when the extracorporeal perfusion was prolonged, the disturbance in the first and the second phase in clotting mechanism were remarkable and persistent to result in voluminous postoperative hemorrhage. Hence, it is quite desirable that the extracorporeal perfusion is terminated within 2 hours after the start. 3) Most of the patients with cyanotic congenital heart disease were accompanied by abnormalities in vascular resistance, count and function of platelet and factors relating to blood coagulation. It is of clinical importance that postoperative augmentation of these abnormalities was not rare. 4) The hemodilution with low molecular weight dextran, modified by the author, was applied for the extracorporeal circulation to obtain profound hypothermia. In this profound hypothermia, such abnormalities in clotting factors as manifest the bleeding tendency were not observed, though heparin rebound phenomena appeared in occasional cases. 5) In the extracorporeal circulation, the application of hemodilution with low molecular weight dextran solution by twenty per cent of the total priming and circulating blood volume was advantageous for prevention of occurrence of the abnormalities in clotting process which was caused by undiluted whole blood.
The incorporation of 1-14C acetate into major lipid fractions or discrete fatty acids of whole blood has been studied. The subjects studied were 22 cases of liver cirrhosis, 11 severe diabetics, 8 mild diabetics, 7 normal persons, and 5 cases of acute hepatitis. Of these 22 cases of liver cirrhosis, 2 were normal, 10 were abnormal in glucose tolerance test (GTT), and another 10 cases were clinically considered to have been complicated with primary diabetes. Results: 1) Not only the groups of liver cirrhosis with abnormal GTT and with diabetes, but severe diabetics showed remarkable decrease in incorporation of 14C from acetate-14C into fatty acids and non-saponifiable fraction, howeuer, little depression has been found in the mild diabetics and the group of acute hepatitis as compared to the normal group. 2) From the view of incorporation of 14C into iudividual fatty acids, groups of liver cirrhosis showed equally less in percentage of recovered cpm in 14: 0, and 16: 0 than those found in diabetic group, particularly, percentage incorporation of 14: 0 in group of liver cirrhosis with abnormal GTT was significantly decreased as compared to diabetic groups. In 16: 0 also, percentage of recovered cmp in both groups of liver cirrhosis with abnormal GTT and with diabetes was significantly depressed than controls. All groups of liver cirrhosis showed highly significant increase of percentage incorporation in 18: 1 as compared to both normal and diabetic groups. Significant increase of percentage of recovered cpm in fatty acids with retention time corresponding to 20: 4 or greater was found in the groups of liver cirrhosis as well as diabetics as compared to normal controls. Therefore, the groups of liver cirrhosis and diabetics showed analogous changes in the formation of 14: 0, 16: 0, and fatty acids longer than 20: 4, but the remarkable increase in formation of 18: 1 was found to be specific for liver cirrhosis. 3) The percentage of recovered cpm in 14: 0, 16: 0, and fatty acids longer than 20: 4 had no significant difference among the groups of liver cirrhosis. However, in spite of insignificant change, the group of liver cirrhosis with abnormal GTT showed increase by about 3% in mean value in percentage incorporation in 18: 1 as compared to the group with diabetes. 4) No particular changes in percentage incorporation of 14C into discrete fatty acids of blood were observed in the group of acute hepatitis as compared with normal group.
Rat epididymal adipose tissue was incubated in the normal human serum in the presence and absence of insulin, and the incorporation of labeled glucose and acetate into major lipid fractions and individual fatty acids was compared as these in Kreb's bicarbonate buffer. The results were as follows: 1) The incorporation of 14C from 1-14C glucose or 1-14C acetate into fatty acids of adipose tissue was increased by the serum containing 50μU/ml of added insulin, but the incorporation was inhibited about 20% by serum with 800μU/ml of added insulin. 2) The percentage of recovered cpm incorporated into various fatty acids showed some difference in the serum as compared to the values in buffer. Although the percentage of 14:0 and 16:1 was decreased and 16:0 and 18:0 was increased by the serum, the changes found in these fatty acids were less sensitive to the effect of added insulin. The formation of 18:1 was most sensitive to added insulin in serum and the change of synthesis of this fatty acid reflects most specifically the stimulatory or inhibitory effect of insulin on the formation of total fatty acids in serum. 3) The effect of insulin as was found in fatty acid formation in serum was also appeared in the formation of non-saponifiable fraction in serum, but the glycerol formation was almost independent on insulin effect. 4) Dialyzed sernm with 200μU/ml of added insulin stimulated the formation of total fatty acids remarkably, but the formation of 18:1 was also depressed in this condition, then, it is conceivable that the formation of 18:1 has some difference in nature as compared to others.
In an attempt to clarify the process of transformation in energy metabolism during the reticulocytes maturation, the changes of acid soluble nucleotides during the maturation in vitro were analysed column chromatographycally and the following results were obtained: 1. The acid soluble nucleotides of reticulocytes were mainly composed of cytidine monophosphate, nicotinamideadenine dinucleotide, adenosine monophosphate, inosine monophosphate, adenosine diphosphate, guanosine diphosphate, adenosine triphosphate and guanosine triphosphate. In such nucleotide components the purine nucleotide compounds were composed of 70 to 80% of whole nucleotides. 2. During the maturation in vitro nucleotides and adenosine triphosphate contents decreases but the ultra-vioiet absorbing materials as a whole increases with the increases of base and nucleoside contents. 3. The changes of acid soluble nucleotides during the maturation were stimulated by 2, 4-dinitrophenol or antimycin A. 4. The accumualation of hypoxanthine during the maturation was stimulated by adding 2, 4-dinitrophenol or antimycin A which inhibited the RNA degradation, accopaning with the degradation of acid soluble nucleotides. 5. The reticulocyte maturation, the RNA degradation is largely dependent on energy from the mitochondrial oxidative phosphorylation.
The present paper reported the relationship between nucleotide metabolism and energy metabolism during the maturation of rabbit reticulocytes and the following results were obtained: 1. The gradual degradation of adenine nucleotides during the maturation of reticulocytes was stimulated by the respiratory iuhibitors, antimycin A and amytal, or an uncoupler of oxidative phosphorylation, 2, 4-dinitrophenol and an inhibitor, oligomyciin. 2. In these conditions the pentose moieties of adenine nucleotides were metabolized rapidly with the very large accumulation of hypoxanthine and lactic acid. 3. The processes of maturation of reticuloc t s, RNA decreases and the glycolysis were promoted by adding inosine and also by inosine monophosphate as well as by glucose. Theexogenous adenosine diphosphate and ribose-5-phosphate were hardly utilized for reticulocyte maturation. 4. The data suggest that pentose moieties of adenine nucleotides are reutilized effectively than glrcose during the reticulocyte maturation.