Chromatographic separation of polar dyes was carefully carried out. The enzyme and the alkali-polar dyes were respectively fractionated into three and two major components on Amberlite CG-50 column. The first and the second fractions of the enzyme-polar dye showed chromatographic behavior similar to the corresponding fractions from the alkali-polar dye. The third fraction of enzyme-polar dye seemed to be greatly decreased in quantity in the alkali-polar dye. Each fraction of the alkali-polar dye was purified by ascending paper chromatography. The second fraction was further separated into two components. Their physicochemical properties such as electronic absorption spectra and ionization constants were determined.

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Enzyme-polar dyes prepared from liver of rats fed rice mixed with 4-dimethylaminoazobenzene for two weeks and fractionated by Amberlite IRC-50 column chromatography using 60% ethanol as the eluant were shown to be still contaminated with a small amount of peptides. Alkali-polar dyes prepared from the enzyme-polar dye treated with ethanolic potassium hydroxide, followed by Amberlite IRC-50 or CG-50 column chromatography, were almost free from contaminated peptides or amino acids. The alkali-polar dye was fractionated into four main components by the combined chromatography using the column and the filter paper. All of these purified alkalipolar dye components showed the presence of one α-amino group and one phenolic hydroxyl group. The amino acid bound to the dye could not be liberated by the ordinary acid or alkali hydrolysis used for peptides or proteins. Only one out of the four components behaved somewhat differently from the other three components in the absorption spectra, reactions with nitrous acid, as well as with 2, 4, 6-trinitrobenzenesulfonic acid and was assumed to have a primary amino group.
Another possible chemical configuration was presented in addition to the old formulae.

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Each of ten male rats, Buffalo and Long-Evans strains, received subcutaneous injections of 4-nitroquinoline 1-oxide. The final effective number of rats was 7 and 8 in the groups 1 and 2, respectively, after 164 days from the start. In most of rats, one or two subcutaneous sarcomas were induced, multiple pulmonary tumors were induced in 13 of 15 rats. Almost all the tumors were designated histologically as adenomas. Two cases of squamous cell metaplasia were found. Further, multicentric carcinomas, adenocarcinomas, and squamous cell carcinomas were induced in 3 rats among them, which survived more than 295 days. Sudden transitional connections between these adenomatous and cancerous growths or bronchi and frank carcinomas were found very frequently. From these evidences, it seemed that pulmonary carcinomas in rats will be derived from either the adenomatous growths or the bronchial terminals.

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1) Immunological reactions in cancer patients and tumor-bearing mice were studied by means of active immunization with tetanus toxoid.
2) The tetanus antitoxin in C3H/Heston mice bearing MH-134 carcinoma showed normal level on the 6th day after reimmunization, but it abruptly decreased on the 12th day.
3) As compared to mean antitoxin levels, the primary response in 28 cancer patients was inferior to that in 23 normal subjects. In the secondary response, however, the cancer group produced an equal amount of antitoxin as the normal group.
4) There was no correlation between the antitoxin level and growth or rejection of the tumor autografts.
5) The antibody production in malignancy has been discussed in the sense of tumor immunity.

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1) The successively subcutaneously transplanted osteogenic sarcoma, which was produced experimentally and forming little or no bone and which seemingly appeared as polymorphic cellor fibro-sarcoma, showed evident osteoid formation in the 13th generation of transplantation.
2) An evident osteoplastic sarcoma developed when a non-osteoplastic sarcoma of the same origin as the above-mentioned tumor was transplanted into the medullary cavity of a Wister rat.
3) Based on the two experiments it was clarified that a sarcoma originating from the bone possesses latent capacity for osteoplasia even if it seemingly has lost osteoplastic ability during successive transplantations, and can develope into an osteoplastic sarcoma under certain circumstances. Definite bone formation of the sarcoma was found when the non-osteoplastic osteogenic sarcoma was transplanted into the intramedullary cavity of the bone.

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Four groups of 7-week-old male Donryu rats, 120 heads in total, were fed with 0.05% of 3'-methyl-4-dimethylaminoazobenzene for 3, 4, 5, and 6 months. Appearance of tumor rats, autopsy findings, and histological studies of the induced liver cancers were made in each case and compared among the 4 groups. On the other hand, primary tumor ascites, and the largest and the second largest tumor nodules were transplanted intraperitoneally to normal Donryu rats and their transplantability was studied and compared with each other. In addition, successive transplantation was attempted and the transplantability and median survival time in days were examined in each case. Throughout these experiments, two apparent facts were observed. One was that the transplantability of the induced liver cancer has a close relation to the amount of the carcinogen given and it was demonstrated that the longer the period of feeding was, the higher the transplantability of the induced liver cancer was. The second was that the transplantability of the largest tumor was 2-3 times higher than that of the second largest. Based on these results, the mechanism of carcinogenesis was discussed.

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Some cytochemical investigations were carried out on the effect of antisera produced in whole cells, free nuclei, and mitochondrial fraction of HeLa cells and upon HeLa cells in tissue culture, in the hope of gaining some critical information for understanding the mechanism of injurious effect upon tumor cells in vitro.
The antisera affected the cells very actively in a dilution of 1:4 to 1:8 with nutrient medium in the presence of a heat-labile complement-like factor. The HeLa cells exposed to the antisera were damaged within 12 hours through the deformation, pycnosis, and fragmentation of nuclei, and the aggregation of cytoplasmic granules, together with the loss of cytoplasmic basophilia. The reaction rapidly became especially marked on the cell surface. After several cytochemical tests, it was found that there occured a disappearance of ribonucleic acid from the cytoplasm of the affected cells, along with a denaturation or condensation of some proteins.
Several enzymatic studies revealed that the HeLa cells treated with antiserum utilized exogenous succinate, but did not metabolize exogenous glucose, and that peroxidase granules made their appearance in the affected cells, this being the sign of cell damage.
The antisera were found to be antigen-cell specific, so far as the present experiments are concerned. They lost their cytotoxic activity by incubation of the serum at 56° for 30 minutes.
The whole cells, free nuclei, and mitochondrial fraction of HeLa cells were all found to be antigenic and almost equally elicited potent cytotoxic antibodies in heterologous hosts.

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A basic protein fraction having toxohormone activities, isolated from rat Rhodamine sarcoma, was subjected to enzymatic hydrolysis with trypsin, pepsin, and pronase, resulting in the splitting of 12.3%, 19.1%, and 57.4% of peptide bonds of the basic protein respectively.
The effect of the hydrolysis on the toxohormone activities of the basic protein was determined by bioassay with rats or mice. Liver catalase depression was markedly diminished by hydrolysis with pronase and pepsin but not by trypsin. Plasma iron decrease was affected by none of these enzymes. Liver tryptophan pyrrolase depression was markedly diminished by hydrolysis with any one of these enzymes.
These results suggest that these three different activities of toxohormone are associated with different chemical entities of the basic protein.

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The antitumor action of SB₂₁ and SB₂₂, the most active compounds among 100 new Sarcomycin derivatives, has been evaluated by their inhibitory action on various types of solid tumors (Ehrlich carcinoma of the mouse; IRE reticulo-sarcoma, Walker carcino-sarcoma and SIRE fibrosarcoma of the rat).
The differential sensitivity of the various types of solid tumors and the importance of the administrative route have been deeply investigated. The effectiveness of the two compounds has been evaluated by the inhibition of the tumor growth, and by morphological and metabolic modifications following the treatment.

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A preliminary experiment was carried out to investigate the carcinogenicity of 4-nitroquinoline 1-oxide in the guinea pig. After receiving maximum of 20 subcutaneous injections of this carcinogen once a week in a dose of 0.25 or 0.5mg, only one animal of the effective nine guinea pigs developed a tumor at the site of injection. The tumor was found about 2.5 years after the start of the experiment and killed the host 5 months later. The tumor was histologically diagnosed as a fibrosarcoma. The result of the present experiment would indicate that the guinea pig is resistant to the carcinogenicity of 4-nitroquinoline 1-oxide.

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4-Nitroquinoline 1-oxide was administered directly into the glandular stomach of rats through a gastric fistula of polyethylene tube set operatively beforehand, followed by painting of 20-methylcholanthrene on the skin of the back. A marked atypical, epithelial growth was induced in the glandular stomach of one among a total of 33 rats (in both A and B Groups). The lesion was an adenocarcinoma according to Stewart's criteria.

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