Canine dystemper virus (Hokkaido strain) could infect to dog kidney cells and adapted to them. Namely, main CPE by this virus infection were multinuclear giant cell formation, cytoplasmic and intranuclear inclusion bodies formation etc. That was pecuriality which it took almost a month to manifest the CPE after inoculation. However adaptaion of this virus to the cells could be established easily 2-3 genelations of subcultures, and showed the stabilized infection titer. HeLa cells, FL cells, JTC-3 cells and PS cells could be infected with this virus, but the dog kidney cells were more suitable for virus infection, and it was late that these infected cells were separated from glaswall of test tubes.
These dog kidney cultured cells were the suitable cells to these experiments of distemper virus.

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Comparative experiments of immunoserological and pathologic-morphological relationship between dystemper and measles viruses in dog kidney cells on the same conditiom of experiments could be done.
Namely, both viruses showed the quite same CPE, which could be difficult to distinguish, but its manifestation period of CPE were different, and measles virus took about a week after inoculation, dystemper virus took about a month. And then, infection titer of measles virus was somewhat higher than of dystemper virus. Immunoserologicaly, measles patients' serum had of course neutral antibody to measles virus, and contained more higher value of neutral antibody to dystemper virus. However, there was not the definite quantitatve relationship between these neutral antibodies.

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Isolation of Cryptococcus neoformans from the pigeon excreta in the Okayama district was attempted. As the result, 8 strains of Cryptococcus neoformans were isolated from 61 specimens collected at 56 different places in the district.
1. Studies on the pathogenicity to mice of 3 isolated strains, A₁, C₂ and C₉, revealed that pigeon strains were less pathogenic than the Kurata strain isolated from human cyptococcosis.
2. In the mice, Cryptococcus neoformans provoked 2 kinds of lesions, either cystic or granulomatous lesions.
3. By way of intranasal inoculation the fungal metastasis to the brain was difficult, whereas intravenous inoculation induced cystic lesions within the parenchyma of the brain instead of meningitis in the most instances.
4. Though the Cryptococcus neoformans was able to be cultured at 37°C, the strains which had been cultured at 25°C was difficult to produce large colonies in the first generation at 37°C.
5. The capsule of Cryptococcus neoformans was large in vivo but it was found to disappear or become thinner on transplantation to a culture medium.

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1. The rabbits of a commercial stock were immunized by the intravenous inoculation of live Cryptococcus neoformans of the Ishino strain at the concentration of 1 mg per cc. The antigen was injected thrice a week for 3 consecutive weeks. The immune serum with the agglutination titer of 1:1, 024 against the homologous organism was obtained 5 weeks after the last inoculation.
2. The tube agglutination test with this immune serum gave the following agglutination titers, 1:128 against Cryptococcus neoformans (C_2-4 strain), 1:32 against Cryptococcus diffluens and 1:8 against Torulopsis sake and Torulopsis pinus, respectively.
3. The slide agglutination test showed that the differentiation between Cryptococcus species and Torulopsis species was made if the antiserum was diluted to 1:10.
4. The γ-globulin concentation of the fluorescein isothiocyanate-labeled antibody solution was 0.52% which was completely able to differentiate between Torulopsis pinus and Cryptococcus species if diluted to 1:8. The fluorescent antibody technic was able to afford us making a diagnosis without difficulty when the fluorescein isothiocyanate-labeled antibody was prepared from the immune serum with agglutination titer as high as possible and diluting it to some extent. (In the author's experiment, diluting to 0.0165% of γ-globulin was most effective.)

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A study on experimental infection of cryptococcosis was carried out through oral, mucosal and subcutaneous inoculations. The results were as follows:
1. Encephalomeningitis was found only in the cases inoculated subcutaneously with mucin in the buccal region. In mice, 7 cases out of 8 (88%) and in rats, 5 cases out of 6 (83%) gave positive results.
2. Among these, meningitis was seen in 6 cases (86%) in mice and in 5 cases (100%) in rats, respectively. In these cases, histopathologic study revealed invasion of Cryptococcus neoformans in the lymphatic spaces of the brain.
3. Among the routes of infection of Cryptococcus neoformans the infection through the lymphatic vessel from the skin lesion of the head or the neck to the meninges proved to be efficient.

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In the experimental infection of Cryptococcus neoformans M₁ strain against the albino rats weighing 80 to 90 grams, morbidity, quantitative reduction culture from the various organs, histopathologic studies and 17-OHCS measurement in 24-hour urine were carried out. The retults were as follows:
1. The rats administered prednisolone only showed an increased C/F ratio (conjugated urinary 17-OHCS/ free urinary 17-OHCS) in proportion to the period of administraion in case the rats were given prednisolone at a dosis of 50μg. This suggested that prednisolone administered abunduntly becomes conjugated in the body rather than remains free. In contrast the rats given prednisolone at a dosis of 0.5μg showed the same C/F value with normal rats.
2. The unfavorable effects of adrenal corticoids on fungal infections were shown by changes of C/F, morbibity and the results obtained by reduction culture. These effects of adrenal corticoids were influenced neither by the quantity of administered prednisolone nor the time of administration.
3. The effectiveness of antifungal antibiotics on experimental cryptococcosis was determined by the day of initial death, morbidity, C/F ratio and the results obtained by reduction culture. The useful antifungal antibiotics were Amphotericin B, Trichomycin and Nystatin in a descending order of effectiveness.
4. In the experiment in which both. prednisolone and antifungal antibiotics were adminstered, the effects of antifungal antibiotics, even if they were strong enough, were diminished by the simultaneous administration of prednisolone.
5. Prednisolone was found to produce the cystic lesion in experimental cyptococcosis, while the addition of antifungal antibiotics reduced the lesion and induced the intense granulomatous changes. In this experiment, the useful antifungal antibiotics were Amphotericin B, Trichomycin and Nystatin in a descending order of effectiveness.

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In Japanese literature only 15 cases of lipoid pneumonia have been found, including this one.
Fifty-eight-year-old female was referred to Okayama Red-cross Hospital because of non-productive cough.
Exploratory thoracotomy with working diagnosis of mediastihal tumor was done and revealed mediastinal type of squamous cell carcinoma in the lung, accompanied by lipoid pneumonia in the right lower lobe.

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This article is to report a case of reticulosarcoma in the breast. It is very rare disease and only ten cases, including this one, have been reported in Japan.
Fity-year-old female was referred to Okayama Red-cross Hospital because of mass formation in the left breast.
Left radical mastectomy was performed with diagnosis of breast canccr, followed by x-ray irradiation and Tespamin administration, But she died of retroperitoneal, cerebral and cervical metastases in the 50th postoperative day.

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An expedition was performed on Miyako Is., Ryukyu, carrying out medical examinations on school-children in May and June, 1965.
The investigations consisted of screening tests for acatalasemic gene carriers and ENT examinations.
Results obtained are as follows:
1. ENT examinations; morbidity to ear diseases of Miyako children was higher than that of Japan proper. On the contrary morbidity to nose and throat diseases of Miyako was lower than Japan Proper.
2. Screening tests; only one hypocatalasemic boy was found out among 10, 083 children examined. Then emergence of hypocatalasemia on Miyako Is. was 0.01%, one tenth of already known Japanese mean value.

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Since there is a considerable individual difference in bone-marrow disturbances induced by the administration of anticancer agents, the author conducted a series of study on human bone marrow by the clinical tissue culture method devised by Hiraki et al. for the purpose to know the susceptibility of bone-marrow in the individuals bearing cancer, prior to the administration of anticancer agent. The anticancer agents used were Mitomycin C, Cyclophosphamide and Chromomycin A₃, and these were added to the tissue culture at varying concentrations. Observations were carried out as to their effects on the growth of bone-marrow in order to find out the tolerance to anticancer agents of human bone-marrow in in vitro. The results of these observations were compared with those clinical examinations such as the extent of bone-marrow disturbances brought about by the clinical application of each of these anticancer agents, histological pictures of the bone-marrow prior to the administration of the agent, the number of peripheral leucocytes, the serum protein contents, and ratio A/G before the administration of the agent. The findings of the present study are briefly presented in the following.
1. By conducting tissue culture of the bone-marrow obtained from noncancer bearing and cancer bearing patients with the addition of various anticancer agents in varying concentrations, the in vitro tolerance of the bone-marrow to these agents was studied.
i) In the case of bone-marrow tissue culture with the addition of Mitomycin C, both the bone-marrow of the patients with stomach-cancer and that of non-cancer bearing patients showed a marked individual difference in the tolerance to the anticancer agents, and the maximum tolerance proved to be about six time that of the minimum.
ii) In the tissue culture with the addition of Cyclophosphamide, likewise the bone-marrow of both non-cancer bearing and stomach-cancer bearing patients showed the difference of as much as three-fold between its maximum tolerance and the minimum. This proves that the range of individual differences in the tolerance is narrower than that observed in the case with the addition of Mitomycin C.
iii) In the tissue culture with the addition of Chromomycin A₃ the range of individual differences in the tolerance to it was about two-fold between its maximum and the minimum with the bone-marrow of stomach-cancer bearing patients while it was about three-fold in the case with non-cancer bearing patient. This denotes as in the case with the addition of Cyclophosphamide that the range of the individual differences in the tolerance is narrower than that with Mitomycin C.
2. The range between the maximum tolerance and the minimum observed in the bone-marrow in tissue culture is widest in the addition of Mitomycin C, followed by the range in the case with Cyclophosphamide and Chromomycin A₃. However, the decreasing rate of the peripheral leucocytes in the patients after the administration of the agent in clinic proved to be highest with Mitomycin C, followed by that with Chromomycin A₃ and Cyclophosphamide. This fact reveals an interesting mutual relationship between the tolerance range observed in the bone-marrow tissue culture just described and the decreasing rate of peripheral leucocytes after clinical administration of these agents.
3. There was recognized a close mutual relationship between the degree of tolerance to the anticancer agents in the bone-marrow tissue culture added with these agents and the decreasing rate of peripheral leucocytes in the patients administered with these same agents. This finding indicates that the bone-marrow tissue culture conducted with anticancer agent prior to its use can predict the tolerance of the bone-marrow to the agent.

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The both qualitive and quantitative changes of leucocytes were seen during extracorporeal circulation. White blood cell count gradually decreased on starting the extracorporeal circulation. Hemogram showed the neutrophilia with the nuclear shift to the left, and appearance of the immature bone marrow cell in peripheral blood. Adhesive property of leucocyte markedly disturbed, ameboid motility and phagocytic property of neutrophiles were impaired, also.
Considering the effect to leucocyte during the extracorporeal circulation, there are three factors, that is heparin, mechanical acting of heart-lung machine and massive transfusion. Heparin made peripheral leucocyte count decreased and adhesive property impaired. Heart-lung machine destroyed the leucocyte. Massive transfusion decrease the white blood cell count by means of dilution. And both impaired adhesive property, phagocytic property and ameboid motility.
Clinically, using agglutination test and antihumanglobulin consumption test, leucocyte antibody were examined in 30 cases performed extracorporeal circulation and 20 cases with preserved blood transfusion in general surgery. Complete antibody was seen in one case (3.3%). incomplete antibody was in 8 cases (26.7%) with extracorporeal circulation. In 2 cases with preserved blood transfusion (10%) incomplete antibody was also seen. Those revealed one massive transfusion might produce the leucocyte antibody.
The effect to blood coagulation of white blood cell or homogenized white blood cell, which were separated after extracorporeal circulation, were experimentally studied. White blood cell depressed the 1st phase of blood coagulation, and homogenized cell depressed the 1st and 2nd phase. Both two promoted the fibrinolysis. Homogenized white blood cell dissolved the fibrin plate, that were not inhibited with antiplasmin agent. These suggest any proteolytic enzyme except plasmin are acting in concert with activator and / or streptokinase-like substance. Inhibitory property of blood serum to fibrinolytic activity of leucocyte was reduced after extracorporeal circulation.

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Repeated skin painting with 20-Methylcholanthrene (20 MC) caused lymphocytic leukemia in 54.2% of RF mice, of which 18.2% was of the thymic type and 36.3% of the non-thymic type.
The mode of development of the lymphocytic leukemia was serially studied. The peripheral blood pictures showed in the pre-leukemie stage a normochromic anemia, leukopenia, lymphopenia, eosinophilia, and reticulocytopenia; in the early leukemic stage, a slight leukocytosis with a few lymphoblasts; and in the progessive leukemiç stage, a hyperchromatic or normochomic anemia, anisocytosis, polychromasia, thrombocytopenia and leukocytosis with many lymphoblasts.
The imprint preparations disclosed an increase of lymphocytes in the lymphnodes and thymus in the pre-leukemic stage. In the early to progessive leukemic stage, there was a marked increase of lymphoblasts in the lymphnodes, spleen and thymus. In the bone-marrow, increare of lymphocytes and lymphoblasts were noted in the early to progessive leukemic stage.
Histological examination revealed a hypertrophie of the lymphfollicles and an increase of lymphoblasts in the spleen and lymphnodes in the initial leukemic stage of the non-thymic type leukemia. In the thymic type leukemia, a similar increase of lymphoblasts occarred in the thymus. In the early to progessive leukemic stage, an infiltration of lymphoblasts was seen in the spleen, lymphnodes, thymus and bone-marrow. In the liver, the infiltration was pronounced in the peripotal and perivascular spaces.
Tissue culture studies indicated a hypoplasia of the lymphnodes and thymus in the preleukemic stage. However, in the early leukemic stage, an acute leukemic growth pattern was exhibited by the spleen and lymphnodes in the non-thymic type while the thymus of the thymic type showed the same pattern. In the progressive leukemic stage, the spleen, lymphnodes, thymus and bone-marrow presented a culture pattern of acute leukemia.
The incidence of lymphocytic leukemia in splenectomiged RF mice painted with 20-Methylcholanthrene was 42.8%, and that of thymectomiged RF mice with a similar treatment was 18.2%.
From these finding, it is suggested that in RF mice painted with 20 MC the spleen, lymphnodes and thymus are the target organs which first manifest injurious effects and become hypoplastic and in the course of repair and regenerative process, hyperplastic changes untimately result in the leukemic proliferation in these organs.

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The incidence of myelogenous leukemia by repeated skin painting with 20-Methylcholanthrene in RF mice was 29.5% and the mode of development was invertigated, The peripheral blood pictures showed a hypochromic anemia, leukopenia and eosinophilia in the pre-leukemic to intial leukemic stage. In the early leukemic stage, there was a slight leukocytosis with a few myeloblasts. The progressive leukemic stage was characteriged by a hypochromic anemia, anisocytosis, polychromasia, reticulocytosis, thrombocytopenia and leukocytosis with an increase of myeloblasts.
The imprint preparations revealed an increase of neutrophllic cell series in the pre-leukemic stage; and an increare of myeloblasts in the early to progessive leukemic stage, The spleen and lymphnodes showed evidence of neutrophilic cell infiltration in the early to progessive leukemic stage.
Histologically the bone-marrow began to show a slight increase of myeloblasts and other neutrophilic cells in the initial leukemic stage. In the early to progressive leukemic stage, a diffuse proliferation of myeloblasts filled the bone-marrow and invaded the surrounding muscles. There was a slight to moderate leukemic infiltration in the lymphnodes and spleen. The liver showed a leukemic infiltration in the periportal, perivascular spaces and around the central veins.
Bone-marrow tissue cultures showed a hypoplastic pattern in the pre-leukemic stage, a transitional pattern in the initial leukemic stage and a leukemic pattern in the early to progressive leukemic stage, The spleen and lymphnodes assumed a hypoplastic pattern in the initial leukemic stage and a transitional to leukemic pattern in the early to progessive leukemic stage. Of the entire skeletal systems, the lumbar vertebrae, femur and acetabulum fist showed hypoplostic which.was followed by a leukemic transformation.

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The method of separation of tumor cell mitochondria (Ehrlich ascites cell and rat hepa toma cell, AH 130) by treating tumor cells with proteinase, by which the mitochondria are retained in a biologically active state, has been reported. The mitochondria thus obtained proved to have the normal activities of oxidative phosphrylation and respiratory control. The respiratery control of these mitochondria was nearly the same as those of liver mitochondria from normal rat but rather low in ADP/O ratio than the later. The tumor cell mitochondria also showed nearly a normal potency in the swelling at phase I and the oxidation-reduction of pyridine nucleotide, but the swelling at phase II which is induced by adding uncoupers such as oleate and Ca⁺⁺, was markedly low as compared with that of rat liver mitochondria.

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The biologically active mitochondria have been isolated from the livers of normal adult rats and mice, and the tumor cells, Ehrlich ascites tumor and hepatoma cells, AH 130. With these mitochondria some observations have been made on the swelling of mitochondria and lipid peroxide formation under aerobic condition by adding ferrous ion, GSH with GSSG and ascorbic acid. The liver mitochondria from normal animal consumed oxygen actively after a certain lag phase, with the increased lipid peroxide formation by adding the ferrous ion. This peroxide formation was increased by the increased ferrous ion concentration, concomitant with the increased oxygen consumption and the swelling of mitochondria which always appeared after the malonaldehyde was accumulated to a certain level. There was a close relation between the amounts of consumed oxygen and malonaldehyde formed. These phenomena, which were also observed by adding GSH with GSSG or ascorbic acid, seem to be closely correlated to the metabolism and the structural changes of mitochondria. The mitodhondria of both Ehrlich ascites tumor cells and hepatoma cells show hardly any detectable changes in peroxide formation and mitochondrial swelling by adding ferrous ion, GSH with GSSG or ascorbic acid, presenting clear differnces in the biochemical and physiological characteristics of the tumor cell mitochondria from those of the normal cell mitochondria.

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The mitochondria isolated from ascites tumor cells have a specific characteristic different from those isolated from normal liver cells, i. e. they do not react with ferrous ion, GSH with GSSG and ascorbic acid, showing no increase in lipid peroxide formation and no swelling. From the viewpoint that this characteristic of tumor cell mitochondria may be due to the specific liqid. compositions of mitochondrial membrane, the author tried to analyze the components of liqids of tumor cells comparing with those of brain, liver, heart, kidney and spleen from normal animals.
The following have been revealed;
1. The tumor cells are very low in peroxide formation in the presence of the reducing agents comparing to those from the normal tissues, though the grade of peroxide formation is different in each tissues and by the agents used, and is rather lower in homogenate than in intact mitochondria.
2. Chemical analysis revealed that the lipids from tumor cells is low in unsaturated fatty acid contents, especially C₂₀-polyene contents than those from normal cells. This will be responsible for the low level in lipid peroxidation of the tumor cells.
3. On the basement of the these results discussions has been made on the lipid peroxidation of mitochondria of tumor cells and related mitochondrial swelling under the consideration of enzymic activities.

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