The mechanism of carcinogenic action of 4-nitroquinoline N-oxide was analyzed based on the assumption that the substitution reaction with SH compound may be the first reaction which this substance undergoes in vivo. Experimental results indicated that this reaction itself, but not any of the chemical products of the reaction, may be the proximate cause of carcinogenesis, leading to the hypothesis that the energy released by the reaction produces misconstruction of the genetic determinant of the somatic cell (duplicant) by disturbing its normal replication process.
This hypothesis was discussed in relation to the universal proximate mechanism applicable to all the forms of carcinogenesis.

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1) DPN synthesis activities in the livers of tumor-bearing animals showed lower level than that of the control.
2) Tumor homogenate, toxohormone and its partially purified fractions inhibited in vivo the DPN synthesis activity of the liver, but this effect was not confirmed in vitro.
3) The rate of ³²P incorporation was the lowest into DPN among the acid soluble nucleotides and the relative specific activity of DPN in the liver of tumorbearing rat was a little lower than that of normal one.

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1) Rats were fed on DAB diet for 90-150 days, and catalase, nonhemin iron, copper, ascorbic acid, and riboflavin were determinted in the rat liver.
2) Hemosiderin iron in the non-cancerous part of liver maintained the level above normal during carcinogenesis, but that in the induced hepatoma was in the normal range.
3) Nucleoprotein-bound iron increased gradually after 3 months' feeding and reached maximum in the hepatoma and non-tumorous part of liver bearing hepatoma.
4) Ferritin iron of liver during carcinogenesis increased at first and then decreased, but again increased after the appearence of small cancer nodules. When large hepatoma appeared, ferritin iron in the non-tumorous part of liver was lowered under the normal. The ferritin iron content of hepatoma had the lowest value.
5) Catalase activity in the cirrhotic part of liver dropped at first, and then approached normal, but it became above normal after the appearance of small cancer nodules. Both hepatoma and non-tumorous part of liver bearing large hepatoma had definitely lower catalase activities than the normal.
6) The copper content of liver decreases gradually during DAB carcinogenesis.
7) Ascorbic acid in hepatoma and in the non-cancerous part of liver is maintained the level above normal during carcinogenesis.
8) Riboflavin in the hepatoma induced by DAB was less than that in the normal liver. During carcinogenesis, however, the lowest riboflavin content was observed in the liver of rat fed DAB for 30 days, and then the content of this vitamin approached normal.

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1) The DPN dependent formaldehyde dehydrogenase activity (FDH) and xanthine oxidase (X.O.) activity of rat and mouse liver were compared. The mouse liver showed several times as high enzymic activities as those of rat liver. This observation was considered to explain at least partly why mouse liver forms much less protein bound dye and is less sensitive to the dye carcinogenesis compared with rat liver.
2) With rat liver, the intragastric administration of the carcinogenic DAB greatly diminished FDH activity but the non-carcinogenic AB had no decreasing effect. The FDH activity of mouse liver was also decreased by the administration of DAB, but to less degree compared with rat liver.
3) X.O. activity of rat liver was decreased by the administration of DAB. MAB showed almost no effect and AB appeared to increase the X.O. activity of both rat and mouse liver. X.O. activity of mouse liver was not altered by the administration of DAB or MAB.
4) The metabolism of N-methyl group of aminoazo dye seems to be responsible for the drastic decrease in FDH and X.O. activities in the rat liver.
5) The evidence was presented to show that the azo group reductase is different from the xanthine oxidase.
6) The meaning of N-methyl metabolism with respect to the protein binding of the carcinogenic aminoazo dyes was discussed.

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The uptake and conservation of ⁶⁵Zn by Ehrlich ascites carcinoma cells were studied by means of the subcutaneous and intraperitoneal injections of ⁶⁵Zn and the labeling of tumour cells by ⁶⁵Zn. The results obtained are as follows.
1. The ⁶⁵Zn injected subcutaneously below the dorsal surface of the tumour-bearing mouse was incorporated into the liver and the other tissues and then gradually transfered into the ascites tumour cells.
2. In the case of intraperitoneal injection, the ⁶⁵Zn was directly incorporated into the ascites tumour cells.
3. In both the subcutaneous and intraperitoneal injections, the ascites tumour cells took up more of the ⁶⁵Zn than the tissues of normal and tumour-bearing mice.
4. The ⁶⁵Zn incorporated into the tumour cells was conserved during a fairly long period, of which the biological half-value period was about 12.3 days.
5. The ⁶⁵Zn in the dead tumour cells was very rapidly excreted from the abdominal cavity of the mouse.
6. The ⁶⁵Zn-excretion from the tissues was delayed in the ascites tumour-bearing mouse.

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1) In the liver of gastric cancer, I-fraction of copper, that is labile copper fraction, increases in some inverse proportion to catalase activity.
2) The same results was not obtained in the liver in cancer bearing animals, such as Ehrlich ascitic mice and Yoshida sarcoma rats.
3) In the liver of gastric cancer patients and in toxohormone treated mice, β-glucuronidase decreases.
4) Neocuproin, which is thought to be a specific copper chelating substance, can inhibit the liver catalase activity lowering action of toxohormone, and the fall of liver catalase activity in tumor bearing animals may also be, to some grade, protected against.
5) This substance, Neocuproin, may be help to clarify the mode of action of toxohormone. Accordingly, copper may take very important role in the liver in gastric cancer. We think it may be of great significance in cancer cachexia.

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The purposes of this experiment are to examine the splenic changes in tumorbearing animals and to clarify the mechanism and pathogenesis of this phenomenon.
Male mice were employed as the experimental animals and Ehrlich's ascites carcinoma as the transplantable tumor. The results are summarized below:
1) The tumor-bearing mouse showed a pronounced increase in the splenic weight and yet a good correlation between the tumor-weight and the splenic weight was noted.
2) The blood of the tumor-bearing animals is believed to contain a factor which has the property of causing splenic enlargement of the host following its intra-abdominal inoculation and this effect can be observed by serial passage through several normal mice.
3) The factor was contained in the blood serum as well as in the blood cells of tumor-bearing mice.
4) The active material was thermolabile.
5) From the DNA analysis of the spleens of mice treated under various conditions, it was suggestive that the factor in the blood of tumor-bearing animals caused the hyperplasia of cells in the spleen and, secondarily, congestion of the organ. The rate of hyperplasia was shown to prallel the rate of tumor growth.

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The present paper describes the procedure of the establishment of the rat hepatoma in an ascites form named ascites hepatoma II, which was induced by administration of p-dimethylaminoazobenzene; notes are presented on the structural features, growth properties, and behavior of hepatoma cells following enzyme treatment. The tumor cells take formation as cell groups called "hepatoma-cell islands". With the passage of serial transfers, the tumor increasingly showed rapid and invasive growth. Along with these changes, there occurred transitions in structural features of the hepatoma-cell islands, in transplantability of the tumor and in the life span of the tumor-bearing animals. Particularly the morphological changes of hepatoma-cell islands in a transplant generation were remarkable.
In early transfer generations, the chromosome number of tumor cells fluctuated within a wide range. In later generations, the variation of the chromosome number became narrow in range, with the modal numbers at 44 to 49. In the recent samplings cells containing 47 chromosomes occurred at the highest frequency forming a stem lineage. The stem-cell of the ascites hepatoma II is characterized by the constant existence of 27 rod-shaped chromosomes and 20V- or J-shaped elements varying in size.
The subcutaneous or intermuscular injections of the ascites hepatoma resulted in solid tumor growths. These solid tumors proved transplantable in series, and after the 20th passage they still transfered into an ascites form developing in the peritoneal cavity. Tumor suspensions of the 30th passage did not produce ascites tumor easily.
The application of trypsin or chymotrypsin effectively served to separate cells in many hepatoma-cell islands resulting in the formation of free cell suspension. The free cell population did not continue to proliferate in a free cell state in the peritoneal cavity, but re-formed tumor-cell islands. Some factors responsible for the cellular aggregation were considered. A free cell line was successfully induced in one of 34 experimental trials with enzymatic digestion. Possibilities for the formation of the free cell line were considered.

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Carcinoma 51 was transplanted subcutaneously into mice of dd-strain. The changes in content of glycogen, RNA, DNA and phospholipids in the livers, as well as alteration in mitochondrial morphology, were studied by a series of cytochemical staining and cytophotometric methods.
The results obtained indicate that: 1) glycogen is gradually depleted in livers of tumor-bearing mice and becomes completely absent when mice were moribund; 2) the DNA-content in liver cells of tumor-mice shows an increase 7 and 10 days after tumor-transplantation, in concomitance with the increase of nuclear volume of liver cells; 3) RNA was found to be rather rich in livers of some tumor-mice; 4) the phospholipid-content of liver cells as revealed by the acid hematein test shows a decrease specially in the centrolobular area of the liver, and 5) filamentous mitochondria in cells of the centrolobular area tend to break down into granules, and loose phospholipids.

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By consecutive injections of mitomycin C and MBAO, sublines of Hirosaki sarcoma resistant to each agent respectively were obtained. A variant developed through the use of mitomycin C showed a cross-resistance to MBAO and TSPA, but not to other unrelated antibiotics. Moreover, a variant developed through the use of MBAO showed a cross-resistance to mitomycin C.
From these results the author concluded that the mechanism of antitumor activity of mitomycin C might be similar to that of alkylating agents.

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The amounts of B vitamins in the urine of rats fed DAB, AB and rice only were determined.
The concentration of thiamine, niacin, and pyridoxic acid in the urine showed no remarkable changes by the ingestion of azo dyes. Riboflavin, however, was amazingly increased in the urine of rats fed DAB. AB had also some flavin ejection effect, but this effect was weaker than DAB. The amount of riboflavin in the urine of guinea pig, which resists tumor induction by azo dyes was not affected by DAB feeding.

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1) Rats were fed on DAB for about one month, and the activities of some flavin enzymes in the liver were tested.
2) D-Amino acid oxidase was damaged seriously by DAB feeding, and its activity was never improved by the in vitro addition of FAD.
3) The initial reaction velocity of xanthine oxidase prepared from DAB-affected rats was not reduced when compared with the enzyme from the normal, but the velocity of the DAB-affected enzyme gradually dropped off during incubation. This deterioration phenomenon was not observed in the normal enzyme. The addition of FAD to the reaction mixture of DAB-affected enzyme had no improving effect on the enzyme activity.
4) Succinic dehydrogenase activity was decreased by DAB feeding, but the addition of FAD to the reaction mixture almost completely restored its activity.

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1. The capability of PN synthesis as influenced by nicotinamide administration is very low in the Ehrlich ascites tumour cell and the liver of its host mouse, as compared with that of normal mouse liver. In the case of the liver of tumour-bearing mouse, the increase of PN after the administration of nicotinamide is half that observed in normal mouse liver.
2. After the administration of cortisone acetate, the activity of PN synthesis in normal mouse liver always decreases to the level of tumour-bearing liver, suggesting some such hormone may be acting in the tumour-bearing animal.
3. However, the already reduced activity of PN synthesis in the liver of mouse bearing ascites tumour and tumour cell itself is hardly affected by the administration of cortisone acetate.

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The rats transplanted with Rhodamine sarcoma were fed on high iron supplied diet with the aim of correcting the symptoms presumably arising from the iron deprived state in the hosts.
Iron supplement in 2mg dose a day adequately prevented the elevation of erythrocyte porphyrin and liver free porphyrin of tumor-bearing rats, but anemia and liver catalase depression of the tumor bearer were corrected incompletely by iron administration.

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The anerobic glycolysis of the cancer cells was inhibited by nitroquinoline N-oxide derivatives, but the extents of glycolysis-inhibition were not closely proportional to the reaction of these substances with the sulfhydryl group. The inhibition of glycolysis by nitroquinoline derivatives was prevented by the addition of nicotinamide, and these compounds decreased the DPN level of the inhibited cell, as was reported in the case of ethyleneimine compounds by Roitt as well as Holzer et al.
The depression of DPN level by nitroquinoline derivatives was also counteracted completely by nicotinamide. The dffects of nitroquinoline derivatives on the DPN synthesizing and splitting enzyme systems of various tissues were tested and it was revealed that the both enzyme activities especially of cancer cells were inhibited. But the grades of these inhibitions were not so dramatic as was of the glycolysis, and the significance of these inhibitions remains as an open question.
Among the sulfhydryl enzymes tested in these experiments, the nitroquinoline N-oxide inhibited the DPN-dependent α-ketogiutarate oxidase but had no interference with DPN-nondependent succinate oxidase and hexokinase activities. But the inhibition of α-ketoglutarate oxidase by nitroquinoline N-oxide was not prevented by the addition of nicotinamide or DPN itself.
From these results it may be concluded that nitroquinoline derivatives inhibit glycolysis by depressing DPN level as in the case of ethyleneimine compounds but not of monoiodoacetate, which latter inhibits glycolysis by blocking sulfhydryl enzyme such as glyceraldehyde dehydrogenase.

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From Ehrlich ascites tumor cells, RNA was extracted according to the method of Kirby, which was designated as 'pRNA'. The residual RNA ('rRNA') was then extracted from the residue by the method of Schmidt-Thannhauser.
The pRNA fraction was very inactive in incorporation of ³²P-orthophosphate, and the specific activity of this RNA fraction was found to be very close to that of DNA; on the other hand, that of rRNA is strikingly higher.
The results obtained seemed to confirm our previous supposition and the difference of the two species of RNA was discussed.

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By the application of a new type of powerful carcinogens, 4-nitroquinoline N-oxide and some of its derivatives, on tissue culture cells, characteristic intranuclear inclusion bodies were produced in the resting cells. From histochemical point of view, these nuclear inclusions may be considered to be depolymerized DNA or basic protein in nature.
The complete coincidence between the carcinogenicity and the ability of the nuclear inclusion formation was observed among 4-nitroquinoline N-oxide and its derivatives and related compounds, but carcinogens of unrelated chemical groups failed to give rise to similar nuclear inclusions.

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1. Correlative study of histological, histochemical and biochemical changes occuring in the mouse skin within 48 hours after a single painting of 4-nitroquinoline N-oxide and other quinoline derivatives is reported.
2. The early histological skin lesions caused by 4-nitroquinoline N-oxide were consistent with the vesicant actions.
3. The occurrence of these skin lesions were associated with the decrease of intraepidermal SH-content in the treated areas.
4. Among a variety of quinoline derivatives, the potencies causing these skin lesions were definitely correlated with their reactivity with SH-group in vitro.
5. These data indicate that 4-nitroquinoline N-oxide reacts with SH-group also in vivo as the first step of its biological action.

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1) At the Fiftieth passage level established human amniotic cells "H" strain, which was originally derived from normal person, had the potentiality to produce tumors on transfer into the x-irradiated and cortisone treated (Toolan's) rat. Similar results were obtained with HeLa cells.
2) Recultivation of tumors or passage into subsequent series of Toolan's rats were made.
3) Immunocytopathogenic effect of anticellular sera attested to the human nature of tumor cells regrown in the culture with positive results.

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The results of observations of chromosomes in tumor cells invading various organs of a Yoshida sarcoma rat were described in the present paper.
New tumor cells of the Yoshida sarcoma which possess a prominently large J-shaped element have developed in the course of transplantations at Misima. These tumor cells gradually increased in number during the course of ensuing generations. In the process of establishing the new tumor subline two types of cells, with the new and the original karyotypes, were observed. Using this tumor material, the facility of invasion of various organs by the tumor cells with two kinds of karyotypes, the new and the original, was examined. The results of observations indicate that infiltrationability of tumor cells into tissue differs according to their chromosomal pattern. The tumor cells with the new karyotype showed a predominant peritoneal proliferation in an ascites or solid form and a particular adaptability to liver tissue, while those having the original karyotype were well-adapted to spleen and lung tissues.

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The chrm. constitution of Yoshida sarcoma cells was observed by the water-pretreatment method and was analysed on microphotographs. The chrm. numbers were 40 in common generally and were in subdiploid range with few exceptions. The prominent V-chrm. with a characteristic appearance, are found 2 in a tumor cell in high percentage and constancy. But they varied in structure and are classified into several types: Type A consists of 2 curved long arms, the longer arm of which is originated from 2nd J-chrm.; Type B is rather J-shaped with a straite short arm and a long arm, the latter originating from 1st J-chrm.; Type C is bent at 1st constriction with 2 approximately equal arms, one of which is originated from 5th J-chrm.; Type D consists of an extremely long telocentric chrm. and a short arm, the former originating from 3rd telocentric chrm.; Type E is a submetacentric chrm. intermediate in size; Types F, G and H are similar in size and in relationship between the 2 arms, representing the small class of V-chrm: Types F and G are characterized by the peculiar formation on one arm, and distinguished from Type H. Type H is most common to be encountered. It is determined without difficulties as one component of chrm. in the prominent V-chrm., but its companion can not be determined immediately. It is also determined with uncertainty both components of V-chrms. other than the prominent V. Those chrm. characterized by their moderate size and telocentricity, are found as a companion of a V-chrm. in any type invariably, though it is difficult to identify each of them as a companion. Centric fusion of these chrms. has been assumed by many in regard to the chrm. constitution of the rat tumor cells.
The author is different from others in opinion and offers a hypothesis as follows: the central damage of moderate sized chrm., viz, a short of chrm. mutation, is essential for malignant cellular change; these chrms. are structurally widely varied through selection of a companion or of modus of fusion, central or terminal, and the number of the affected moderate sized chrms., will determine the number of the characteristic V-chrms. against the wide morphological variations.

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1. Effects of 110 selected compounds tested against the Friend mouse virus leukemia are reported.
2. Mitomycin C, triethylene melamine, 1, 9-di(methanesulfonoxy)-nonane, and myleran had a destructive effect on the Friend virus leukemia as seen in spleen weight, bioassay and increase in survival time.
3. Urethane, p-phenylene bis(diethyleneimido phosphate), 6-mercaptopurine, 6-mercaptopurine riboside, thioguanine, 6-(1'-methyl-4'-nitro-5'-imidazolyl)-mercaptopurine, 6-(2, 2-dimethylhydrazino)-purine, purinyl-6-thiocyanate, 9α-fluoro-2α-methylhydrocortisone acetate, estradiol, 3, 3-dimethyl-1-phenyltriazene, p-(diethyltriazeno) benzenesulfonamide, and netropsin had a markedly inhibitory effect on the Friend virus leukemia.
4. Of the better-known materials HN₂, triethlene thiophosphoramide, purine-6-thioglucoside, thioguanine riboside, 5-fluorouracil, thymine, diethylstilbestrol, azaserine, actinomycin C, 8-nitro-1-methyl-2-quinolone, 4-nitroquinoline-N-oxide, and 6-aminonicotinamide had a moderately inhibitory effect on the Friend virus leukemia, and A-methopterin, 2, 6-diaminopurine, 8-azaguanine, 5-fluorouridine, 5-fluorodeoyuridine, colchicine, cortisone, sarkolysin, chloramphenicol, 6-diazo-5-oxo-L-norleucine, fumagillin, actinomycin D, podophyllotoxin, ethylenediamine tetraacetic acid, zymosan, potassium arsenite, and H₂O₂ had practically no inhibitoy effect.

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The antitumor activities of oximes were examined on Ehrlich ascites carcinoma and Crocker sarcoma 180. 1) Levulinic acid oxime showed a clear antitumor activity at the in vitro test, and a slight inhibitory effect in vivo. This oxime was found to show the destructive actions on cell membranes of the carcinoma at the concentration of 20mg/ml, and to inhibit the activity of dehydrogenase at the concentration of 5mg/ml. The necessary dose of the oxime to destroy the selective permeability of the cell membranes was about five times that of nitromin, and about the same with that of sarkomycin. The LD₅₀ of the oxime was 2040mg/kg (mice, intravenously). 2) The fact that γ-aminovaleric acid and levulinic acid did not show the activity suggested the necessity of the isonitroso group of levulinic acid oxime for the display of the activity. 3) About 20 monoximes and others having structural relationships with levulinic acid oxime were examined for their activities, but none showed the inhibitory actions on the tumors in vivo.
4) α-Ketoglutaric acid oxime was found to be a chelating agent and to form chelates with copper, iron, and nickel at the physiological pH. The injurious action of the oxime on the selective permeability of cell membranes of the tumor was increased in the presence of copper and iron in the forms of its chelate compounds.

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The chelate compounds of oximes and carbonyl compounds were examined for their antitumor activities on Ehrlich carcinoma and Crocker sarcoma 180.
The activity of the copper dimethylglyoxime was discussed in the terms of ligand and metal moieties, or the extracellular and intracellular activities. The possibility of preparing other active copper chelates, having the selective inhibitory actions on tumor cells, by changing the ligand moiety, was suggested.

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Small liver slices were taken from patients during gastric operations and the catalase activity of the liver was estimated. In gastric cancer a decrease of the catalase activity in liver was generally noted, particularly in those cases in which diffuse atrophic changes in the gastric mucosa were present.

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Five groups of Japanese common albino rats received 0.06% DAB-diet from 1 to 5 months respectively, and, after that, each group was divided into control and experimental groups. After the stop of the feeding, the rats in the experimental groups were painted twice a week, with a 20-methylcholanthrene solution in acetone, and the treatment continued until the 9th experimental month.
(1) The liver findings of the rats sacrificed immediately after the stop of the feeding, (2) the development of tumors and the histopathological liver change, of the rats which died or sacrificed between the 210th and 420th experimental day, and (3) the transplantability of the induced tumors were examined. The results were as follows:
I) If DAB-feeding was continued until liver nodules which showed the histological picture of adenomatous hyperplasia were produced, adenomas and liver cancers could be developed, even when the animals were fed only the basic diet after the stop of DAB-feeding.
II) If DAB-feeding was stopped before the above nodules were produced, and the animals were left without any treatment, no tumor developed. However, if MC-painting was added after the stop of DAB-feeding, some of the nodules were surely developed to liver cancers. But, in these cases, the cancer formation rate was lower than in the groups which received an additional DAB-feeding.
III) The longer the term of DAB-feeding of a group was, the higher was the transplantability of the induced tumors and the more frequent the rate of metastasis in the original animals. MC-treatment had no influence on these points.
On the basis of the above results, the concept of two-phase mechanism on azodye carcinogenesis, and the relation between the doses of applied carcinogens and the autonomous proliferating potency of induced tumors were discussed.

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The technique of diethylaminoethyl-cellulose (DEAE-cellulose) columnchromatography was employed for the fractionation of the raw toxohormone as well as its methanolic acetic acid extract. Under these experimental conditions, the almost all of the original activity was found in the fast moving fractions, equally in the both cases.
Thus, the possibility was suggested that the active components originally present in the raw toxohormone are extracted and concentrated in the methanolic acetic acid extract. The active fractions were demonstrated to be composed of polypeptide. The most active toxohormone was obtained as alkali insoluble components from F-2 fraction, which was effective in 500μg dose per mouse as protein, and its amino acid composition was examined by two dimensional paper chromatography after acid hydrolysis.

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1) The desaminase activity of fatty acid amide in the hepatic tissue homogenate of rats fed with 4-dimethylaminoazobenzene was discussed according to the patterns of enzyme activity, which has been obtained by the method of paper electrophoresis. Moreover the heat-inactivation of the enzyme has been examined.
2) Whole homogenates of hepatic tissues except hepatoma showed an activity pattern with a sharp peak on the starting line and a low convex curve at the fast moving part of protein zone. In case of hepatoma the marked peak on the starting line of the pattern was almost imperceptible. In contrast to this all supernatants of hepatic tissues including hepatoma showed two low curves in the patterns, one at the slow moving part and the other at the fast moving part.
3) Heat-inactivation test to the enzyme on the paper strip has been measured. Whole homogenate of the normal liver lost the prominent peak on the starting line of the pattern, whereas that of the hepatoma the pattern remained almost unchanged even after heat-treatment.
4) Heat-inactivation test of the enzyme was extended to whole homogenates of both normal liver and hepatoma. The enzyme in hepatoma was far more heatstable than that of normal liver.
5) The author postulates to classify the enzyme at least in two sorts, i. e., heatunstable desaminase I and heat-stable desaminase II. The former predominated in the activity of the whole homogenate of normal liver and the latter was represented in the whole homogenate of hepatoma.

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1) Using histamine and cadaverine as substrates, the activity of diamine oxidase in the liver and the other organs of 4-dimethylaminoazobenzene (DAB) fed rats have been measured as compared to those of corresponding organs of normal rats. The activity of tyramine oxidase was also examined as an example of monoamine oxidase. The similar studies have also been carried out concerning the partially hepatectomized and sham-operated rats.
2) The activity of diamine oxidase in hepatoma was similar to that of normal liver. The activity of the enzyme in pathological but noncancerous liver were increased over that of normal liver. Whereas the activity of tyramine oxidase in hepatoma was remarkably low, that in the liver of uneven surface and cirrhotic liver did not so differ from that of normal liver.
3) The rats with cirrhotic liver had a high level of mono- and diamine oxidase activity in the kidneys, pancreas, brain, heart, spleen, etc. The organs of hepatoma-bearing animals showed usually noteworthy height of activity, conspicuous example being the adrenals, when histamine oxidase was tested.
4) The liver and other organs of DAB rats in their early experimental days showed low mono-and diamine oxidase activity compared with those of normal rats.
5) The liver and other organs of partially hepatectomized and sham-operated rats showed the similar tendency of the enzyme activity to that described in the above lines (4).

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(1) An analytical method for the separation and the determination of sugar phosphates developed by the author was applied on testis and malignant tissues in order to get more exact informations of the early stages of sugar utilization.
(2) It is still obscure whether testis glucokinase phosphorylates C-1 or C-6 position of glucose, because of the rapid formation and rapid disappearance of G-1-P in this tissue homogenate. The testis appears to have specific C-6-fructokinase.
(3) Yoshida ascites tumor cells may contain C-6-glucokinase and C-6-fructokinase, and their fructose utilization was quite similar in behavior to that of the testis.
(4) Human mammary carcinoma contains C-6-fructokinase.

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The present paper describes the cytological characteristics of a free-cell hepatoma line which was produced from the chemically induced rat hepatoma of an island-type following transfers in hybrid rats.
The original ascites hepatoma II of rats had been maintained as island-type for 59 transfer generations in pure Buffalo rats from which the primary tumor was originated. After transplantation of this line into hybrid rats between Wistar-King A and Albany, tumor cells composing hepatoma-cell islands gradually lost their cellular adhesiveness and finally produced the free-cell population giving rise to a new subline, AHF.
Comparative studies have been made of the morphological and cytological characteristics of the original tumor, AH-II, and its subline, AHF. It has been shown that the lethal effect, transplantability and growth capacity of the AHF line are stronger than those observed in the AH-II line. Further, the tissue of the AHF line showed in part a sarcoma-like architecture that has not been seen in the tissue of the original AH-II.
The ideogram analysis indicates that the AHF clearly differs from the original AH-II both in chromosome number and other morphological aspects.
There is a possibility to show that the derivation of the AHF line seems to deal with mutational changes in tumor cell (or cells) of the original AH-II. Probably the mutated cells with genotypic changes might be superior in their competitive ability to others and give rise to the formation of the new subline.

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The present article deals with histological and cytological studies of the EM tumor of mice, spontaneous in origin and transplantable in both ascites and solid forms. Originally the tumor developed subcutaneously in the mammary area of a male mouse of EM strain. Through subcutaneous and intraperitoneal inoculations of cell suspension obtained from the primary tumor into new mice, both the solid and the ascites tumor lines were established which have been maintained for 85 transfer generations in the solid line and 130 transfer generations in the ascites line.
Histo-pathological examinations revealed that EM tumor of the solid line was a type of mammary adenocarcinoma, showing malignant growth at the site of transplantation. Temporary smear preparations showed that the tumor cells in the ascites line were generally similar than those observed in most of the other ascites tumors of rats and mice.
The results of the chromosome study of the present tumor in ascites form showed that a numerical shift of the chromosomes from 66 to 67 occurred in the stem cells during the course of serial transfer generations, while the number of 67 was found as a stable stemline number in the later generations. The ideogram of this tumor is clearly distinguishable from the normal ideogram in that it shows a constant occurrence of four large J-shaped chromosomes, a V-shaped element of remarkable size and the remaining 62 rod-type elements. Minute chromosomes, one to three in number, occurred in samples from earlier transfer generations, but disappeared in those from later generations. Generally they take part in the variation of the chromosome number around the modal number.
Lethal transplantability of the present tumor was of more than 98 per cent occurrence in the ascites line and 95 per cent in the solid one. The life span of the animals bearing EM tumor in the ascites line was 10.5 days on the average, while that of the animals bearing the solid one varied rather widely from 6 days to more than 7 weeks with a mean life span of 14.2 days. No significant difference could be detected between EM mice and foreign strain mice such as SM, C 57 BL and D-240 mice with respect to transplantability and life span. Neither was appreciable difference found in transplantability of the tumor during the course of transitional changes of chromosomes.

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The present paper describes the results of a chromosome analysis in tumor cells of a human gastric carcinoma. The samples for study were obtained from the peritoneal fluid of a patient.
It was found that there were present in this tumor three populations of tumor cells which were characterized by the chromosome numbers of 42, 45 and 80-84, each showing a particular chromosome pattern and a high frequency in occurrence. They are regarded as stem-lines of tumor cells which contribute principally to the growth of this tumor: the lines formed by 42- and 45-cells are considered to be principal lines and the line of 80-84 cells is supplementary.
Evidence was obtained that the frequency in occurrence of the two principal stemlines varied in three samplings made on different dates. The variation seems explicable as due to the adaptability of the cell-types to certain physiological conditions of the patient.

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Twenty-seven cases of gastric cancer were studied with respect to frequency of spread into the duodenum and mode of invasion. In supplementary experiments through rabbits, the extension of Indian ink injected into the gastric wall to the duodenum was presented.
1) The duodenal spread of cancer cells was noticed in 5 out of 12 cases of pyloric cancer, and in 4 of the 5 the spread was recognized grossly. In 2 out of 5 cases of cancer of the body, in 2 out of 3 cases of the cancer of the cardia, and in 1 out of 2 cases of the diffuse cancer, microscopic invasions of cancer cells into the duodenum were noticed. In those cases peripyloric lymph glands were enlarged and congromerated by the metastatic involvements of cancer. In 3 out of 5 cases of gastric cancer with gastrectomy, the oral stump of the duodenum was microscopically invaded with cancerous lesions. Macroscopic types of those gastric cancers were ulcerative or diffusely infiltrative. Histological types of the gastric cancers had little relation to the frequency of the duodenal spread of the cancer. Those invasions of cancer cells were chiefly noticed in the lymphatics within the duodenal wall, particulary within the subserosa or submucosa.
2) On the experiments on rabbits, in which fairly large amount of Indian ink was injected into the gastric wall near the pyloric ring it reached the subserosa or submucosa of the duodenum for about 1cm distal from the pyloric ring, but when small amount of Indian ink was injected in the similar portion its extension into the duodenal wall was not recognized. Therefore it is of interest that the physiological lymph flow in the pyloric region may be easily disturbed by a trifling factor.
The above results may suggest that certain disturbances in lymph flows in gastric wall play an important role in the duodenal spread of gastric cancer, and that these spread are chiefly caused by lymphatic permiation of cancer cells or continuous extension of cancer cells through the lymphatic spaces.

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1) Glycine incorporation into the Ehrlich ascites carcinoma cells was studied quantitatively after ¹⁴C-2-glycine injection with the aid of track autoradiography.
2) The autoradiographic method was discussed, especially the quantitative track autoradiography.
3) Glycine incorporation into the interphase tumor cells was rapid, its maximal peak occurring at least 30 minutes after injection.
4) As to the distribution of radioactivity, a rapid decrease in the early period and almost the same level in subsequent period were observed. The former was presumably ascribable to the exchangability of glycine and the latter to the behavior of glycine incorporated into nucleic acid, protein or the precursor.
5) Glycine incorporation into tumor cells, i. e., into their nucleic acid, protein or precursor, took place mostly in the nucleus, and the nuclear membrane played a certain important role, but it may occur also in the cytoplasm in part.
6) Two different types of tumor cells could be observed for their ability of incorporation. Cells having rich chromatin nucleus and deep stained cytoplasm showed an intensive uptake, while cells having poor chromatin nucleus and less stained cytoplasm were inactive.
7) Considerable quantities of the tracks could be observed in the degenerative tumor cells and some of the non-malignant cell fractions.
8) The dividing tumor cells showing uptake were observed and related problem was discussed.

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Oncotrephin was isolated from experimental hepatoma (AH 130) of rat. It can be precipitated not only by 30-70% ethanol, but also by 50-70% saturated ammonium sulfate. The question of the identity of oncotrephin with the growthpromoting factors in the liver (Teir et al. Paschkis et al. and Martel et al.) was discussed.

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Oncotrephin, isolated from a rat ascites hepatoma (AH 130), was added to the culture medium, and its growth-promoting effect, at optimal concentration, for strain L cells was reconfirmed. Neither heating for 30 minutes in boiling water bath nor dialysis abolishes the growth-promoting effect of oncotrephin upon strain L cells.

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