The production of experimental tumor was attempted in 60 C57BL mice by giving them20-methylcholanthrene pellets into the cerebrum and the cerebellum. Tumors were produced in 26 animals-glioma in 3 and sarcoma in 23. These 3 cases of glioma, 2 of the 23 cases of sarcoma and other 2 cases of serially transferred glioma (7 cases in total) were cultivated through succesive generations, and cultures of the 1st, 5th, 10th, and 15th generations, were observed by light and electron microscopy. Light microscopically, all the primary gliomas presented a blastoma-like histological picture. Electron microscopically, constituent cells of the transplanted gliomas each had an irregular formed nucleus, and, in the cytoplasm, many ribosomes as well as abundant cytoplasmic organelles such as ergastoplasms and mitochondria. With prolongation of the serial transfer, however, cytoplasmic processes were lost, cell membrane became smooth, and each constituent cell thus came to exhibit a monotonous fine structure. There were scarcely any charactristic features such as gliafilament which indicate gliogenicity and which are foundin normal or reactive astrocytes or in human astrocytoma.
The constituent cell of the transplanted sarcoma, which developed ergastoplasms, distended cisternae and aboundant intercellular collagenous fibers in closè relation with cell membrane, manifested the charactristic feature of sarcoma, clearly distinguishable from that of glioma.In the glioma of serial transfer through mice, round virus-like particles of double shell structure, about 120mμ in diameter, were observed in ergastplasms and other cytoplasmic organelles; they were also seen in spore-like forms on the cell membrane or accumulated in the intercellular space.
It was demonstrated dy electron microscopy as by light microscopy that the trans-plantable experimental glioma consisted of atypical tumor cells poor in gliogenous features

View full abstract

1. Mice were inoculated with either living organisms or chrome-alum vaccine of Salmonella enteritidis, and sacrificed at each given time to investigate hepatic lesion by electron microscopy and fluorescent antibody technique.
2. Both the living and killed organisms were mainly found in cells of hepatic sinusoids but not in hepatic cells.
3. Proliferation of the living vaccine was seen in Kupffer's cells.
4. Tne killed vaccine underwent marked morphological changes at 5 to 7 days after inoculation, and some were observed in groups.
The living vaccine was altered not only morphologically but also quantitatively, showing transient decrease. But after 7 hours following the infection with living vaccine, they were multiplied to be finally released into blood stream.
5. The fluorescent antibody technique could demonstrate their organisms in the killed vaccine nodules.
6. There was no morphological change in phagocytosis of Kupffer's cells against the living and killed organisms.

View full abstract

⁵⁹Fecl₃ were added to Ehrlich ascites cancer cell solution, the mixture were incubated in a water bath with shaking, and after certain times the uptakes of ⁵⁹Fe by cancer cells, and effects of incubation temperature and of addition of carriers, metabolic inhibitors, amino acids, chelating agents, anti-cancer drugs and metal ions were investigated.
1) As for intracellular distribution of the taken up ⁵⁹Fe, it was found in the greatest amount in the nuclear fraction, next in the supernatant and the microsornes. In the mitochondrial fraction, it appeared in the least amount.
2) Effects of incubation temperature : Between 16°C and 37°C, effects of temperature on the ⁵⁹Fe uptake rate could scarcely be found.
3) By increasing cell number from 5×10⁶ to 20×10⁶ and 80×10⁶, the ⁵⁹Fe uptake rate increased, but the value per cell decreased contrarily.
4) On the addition of Fe³⁺ (FeCl₃) as a carrier, the ⁵⁹Fe uptake rate showed significant increase at 10^-3M, 10^-4M. On the addition of Fe²⁺ (FeCl₂) as a carrier, the ⁵⁹Fe uptake rate showed significant decrease at 20 minutes and increase at 60 minutes. When Fe²⁺ and Fe³⁺ were respectively added to 10^-6M, no significant changes could be found.
5) Effects of addition of metabolic inhibitors : The ⁵⁹Fe uptake was remarkably inhibited at 10^-3M, of DNP, MIA and KCN, using 0.9%NaCl or Tris buffer as a medium, and at 10^-4 M-10^-5M the effect was not so much great. Only by addition of KCN to 10^-3M, using KRP as a medium, the uptake was contrarily promoted.
6) Effects of addition of various amino acid using 0.93/4NaCl as a medium were as follows : At 10^-3M, 10^-4M and 10^-5M of L-Cysteine, 10^-3M of L-Asparti acid and 10^-3M of Na Glutamate, the uptake showed significant increase. At 10^-3M of D-Penicillamine and L-Glutathione the significant decrease was observed. Addition of DL-Penicillamine, Glycine-Na and L-Histidine did not elicit any significant changes.
7) As for addition of various chelating agents using 0. 9%NaCl as a medium, the addition of Ca EDTA (10^-3M) and Dihydrothioctic acid (10^-3M) significantly decreased the uptake rate at 20 and 60 minutes. When Na citrate (10^-3M), Tetracycline (100μg), Ascorbic acid (10^-3M) and Riboflavin (200μg) were added however, the significant decrease was found only at 20 minutes, and when α-thiolactoyl glycine Na was added, no significant decrease could be found both at 20 and 60 minutes. The inhibition rate of ⁵⁹Fe uptake rate by chelating agents including amino acid did not agree with the stability constant.
8) The effects of addition of various anti-cancer agents at various concentrations were as follows : 10μg of Toyomycin at 20 and 60 minutes, 100μg of Merphyrin at 60 minutes significantly decreased the uptake rate, however, 100μg of Nitromin at 20 and 60 : minutes conversely significantly increased it. As for the effects of Mitomycin C and Tespamin, any significant changes could not be found.
9) When various metal ions were added using 0.9%NaCl and Tris buffer as a medium, the degrees of uptake were varying without showing any regularity.
10) Addition of Na ATP, G-strophantin did not elicit any changes in ⁵⁹Fe uptake rate.

View full abstract

Using rabbits administered ³²P and ⁵⁹Fe simultaneously or ¹⁴C-2-glycine, experimental studies were carried out on the relation between erythropoiesis and nucleic acid metabolism in the bone marrow and the following results were obtained :
(1) Results obtained in preparatory experiment revealed that radioactivity due to contamination with ³²P was not detected in hemin fraction from blood or the bone marrow. No contamination was demonstrated with ⁵⁹Fe in DNA and RNA fraction extracted from the bone marrow. There was no free ⁵⁹Fe in the hemin fraction and no free ³²P in the nucleic acid fractions from the bone marrow.
(2) Rabbits were given ⁵⁹Fe-globulinate 6 hours and ³²P 4 hours before sacrifice, and hemin and nucleic acids were extracted from their blood and the bone marrow obtained at the time of sacrifice. Two to four rabbits were sacrificed 6, 12, 24, 48 and 96 hours after a single intraperitoneal injection of anemic sheep plasma with high erythropoietin activity (AP) to determine specific radioactivity of the hemin and nucleic acids extracted from peripheral blood or the bone marrow. Prior to sacrifice, each rabbit was given ⁵⁹Fe and 32P as described above. Specific radioactivity of hemin from the bone marrow was evidently elevated in the rabbits sacrificed 6 hours after AP injection and it remained high as late 96 hours after the injection. Specific radioactivity of hemin from peripheral blood was remarkably lower than that from the bone marrrow, but it was increased from 12 hours after AP administration, attained a peak at 48 hours and remained higher than control value at 96 hours. Specific radioactivities of both DNA and RNA were increased significantly from 12 hours following AP injection and remained in high level until 48 hours.
(3) Similar results to those in rabbits with simultaneous use of ⁵⁹Fe and ³²P were obtained in the rabbits administered ¹⁴C-2-glycine 4 hours prior to sacrifice.
(4) Specific radioactivity of RNA was higher, although it was evidently lower as compared with that in healthy nomal rabbits, in rabbits sacrificed 24 hours after bilateral nephrectomy (Group A) than that in rabbits sacrificed at 54 hours after the operation (Group B) However, specific radioactivity of DNA was equal and remarkably low in these 2 groups of rabbits. In Group A, AP injection immediately after bilateral nephrectomy resulted in significant increase in specific radioactivity both of hemin and RNA. There was no change in specific radioactivity of DNA. In Group B, AP injection 24 hours after the operation resulted in no significant change in specific radioactivities of both hemin and nucleic acids.
(5) On the basis of the above described results, relation between erythropoiesis and nucleic acid metabolism in the bone marrow was discussed.

View full abstract