Changes in the degree of fluorescence polarization in the fluorescent cytoplasm of lymphocytes from patients with malignant diseases, patients with nonmalignant diseases, and normal donors were investigated by the fluorescence polarization technique.
In order to confirm Cercek's cancer test, two kinds of parameters, the degree of fluorescence polarization of lymphocytes stimulated with phytohemagglutinin (PHA) and cancer basic protein, were obtained for each person. Patients with malignant diseases were easily differentiated from the others without exception by an index, the response ratio of the degree of fluorescence polarization.
Usefulness of this technique was confirmed for the detection of cancer-bearing patients.

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A carcinostatic protein fraction (EPF) was separated from Escherichia coli 055 cells by the following procedure: E. coli 055 cells were suspended in distilled water, and extracted by repeated freezing and thawing. The extracts were centrifuged and the supernatant was treated with 1N acetic acid (pH 4.5). Acid precipitate collected by centrifugation was dissolved in alkaline water (pH 8∼9) and subjected to gel filtration on Sephadex G-200 column. The high molecular weight fraction obtained by gel filtration was collected, freeze-dried, and designated as EPF.
EPF was composed of approximately 74.5% of protein, 20.4% of lipid, and a small amount of polysaccharide. The isoelectric point of EPF was pH 5.2±0.1 and the ultraviolet spectrum showed absorption maximum at 278nm (E^1%_1cm 4.30) in 0.01M Tris-HCl (pH 7.8) buffer.
The antitumor experiment with EPF demonstrated that the daily injection of EPF in a dose of 10μg/mouse inhibited sarcoma-180 ascites in 90% of the treated animals and the dose of 100μg/mouse brought about 100% inhibition. In the contact test using C3H2K and C3HW2K cells, 200μg/ml of EPF exhibited about 40% cell destruction. In the inhibition test of ¹⁴C-phenylalanine incorporation into intact sarcoma-180 cells, EPF exibited 60.5% inhibition at the concentration of 275μg/ml. In contrast, EPF did not show any inhibition of the incorporation of ³H-uridine or ³H-thymidine.

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Ehrlich ascites tumour, administered intraperitoneally 18hr following a dose of CCl₄ by stomach tube, produces an irreversible illness characterised by failure to resolve extensive hepato-renal necrosis. Equally extensive hepato-renal necrosis occurs in animals given CCl₄ and saline; such animals, however, remain clinically well and show rapid histological regeneration of both organs. Carbon tetrachloride given to mice on day 5 of tumour growth produces hepatic necrosis only, the kidney of such animals being immune to the necrotising effect of CCl₄. Such animals remain well, and histological recovery of the liver is rapid. It is proposed that the Ehrlich tumour produces a "regeneration-inhibiting" toxin, active against the damaged liver and kidney; the CCl₄-damaged kidney fails to excrete this toxin, hence the irreversibility of hepato-renal damage and a fatal outcome.
A marked reduction of ascites volume as compared with controls was observed in those animals given Ehrlich tumour 18hr after CCl₄, but not in animals given CCl₄ on day 5 of Ehrlich ascites tumour growth.

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Mice of F₁ hybrids of (C57BL/6×WHT) strains were exposed in utero to 200R of X-rays at 12 or 16∼18 days post coitum. Mice of both sexes were allowed to live through their life span, and the induction of tumors, growth retardation, and induction of degenerative diseases were examined. A significant enhancement of lung, pituitary gland, and ovary tumorigenesis was observed in mice irradiated at the late intrauterine stage. Also, incidence of liver and skin tumors was increased slightly in this group, whereas thymic lymphomas were not induced. Persistent growth retardation, several congenital malformation, and amyloid degeneration were found in mice irradiated at the middle intrauterine stage. However, no increased incidence of tumors was seen in this group. Moreover, incidence of lymphoreticular tissue tumors, lung tumors, and leiomyosarcomas of uterus was significantly decreased by irradiation at the middle intrauterine stage from the unirradiated control and the late intrauterine irradiated group.

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A clinical trial of chemoimmunotherapy using cell-wall skeleton of BCG (BCG-CWS) was conducted in 28 patients with acute leukemia in complete remission. Chemotherapy consisted of monthly intensification therapy for 2 months and bimonthly thereafter. Immunotherapy with 200μg of oil-attached BCG-CWS mixed with 10⁷ autochthonous leukemic cells was given intradermally at either of upper or lower extremities every week, except when the patients were on maintenance therapy. No serious systemic side effect attributable to BCG-CWS was noted. Six patients developed mild and transient temparature elevation. Most important side effect was local skin reaction. Indulated papules developed in all patients, resulting in draining ulcerations in 26 patients. Increase of immunological reactivity of the patients receiving BCG-CWS was noted. Skin test response to PPD, Varidase, and candida extract showed definite increase. In vitro lymphocyte blastogenic response to PPD, concanavalin-A, and pokeweed mitogen revealed significant increase.

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The structure-activity relationship in different sensitivity of AH-13 and L-1210 to nitrosourea and related derivatives was examined. N-Methyl-N-nitrosoureido derivatives, such as 1-methyl-1-nitrosourea (MNU) and 1, 1'-polymethylene-bis(3-substituted 3-nitrosourea), were inactive against AH-13 and slightly active against L-1210. On the contrary, 1, 1'-polymethylene-bis(3-substituted 1-nitrourea) derivatives were more active against AH-13 than against L-1210. The nitrosoureas which had bis(2-chloroethyl) group at the terminal were highly active against AH-13 and L-1210. One of denitrosated derivatives, 1, 1'-ethylenebis [3-(2-chloroethyl)urea], was active against AH-13 alone.
In addition, diisocyanates, nitrourea, ammonium carbamate, and 1-methyl-1-nitrourea, which are related to the nitrosourea compounds, were also tested for their activity against AH-13 and L-1210. Diisocyanates and nitrourea were active against AH-13 alone, while other compounds were all inactive.

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The repair of potentially lethal damage after administration of bleomycin by a single injection or by continuous infusion was studied in vivo. Experimental tumors were the fifth generation isotransplants of a squamous cell carcinoma which arose spontaneously in a C3Hf/He female mouse. Bleomycin was administered intraperitoneally by a single injection or by 24-hr continuous infusion. Cell survival was assayed by TD₅₀ method. Dose-survival curve after a single injection exhibited a biphasic or upward concave curve. Surviving fraction increased rapidly if tumors were left in situ after treatment and reached a plateau at 5hr, indicating that tumor cells were able to repair the potentially lethal damage induced by bleomycin. Dose-survival curve after continuous infusion was also biphasic, but had a small shoulder. The repair of potentially lethal damage was slight, if any, when tumors remained in situ for 6hr after the end of 24-hr continuous infusion. This indicates that potentially lethal damage was being repaired during drug infusion.

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Peripheral blood lymphocytes from healthy donors, cancer patients, and noncancer patients were cultured with or without phytohemagglutinin (PHA) for 3 days, supernatant containing lymphotoxin (LT) was added to L cells, and LT activities were examined. LT release by PHA-stimulated lymphocytes from patients with uterine cervical cancer in stages 0 and I decreased and the degree of the decrease was much more marked in stage IV. However, LT activities released spontaneously without PHA were higher in patients with uterine myoma, patients with uterine cervical cancer in stages II to IV, patients with pulmonary tuberculosis, and patients with gastro-intestinal cancer. Relation of LT release and blastogenesis following stimulation with PHA was also examined. Correlation was observed in only 14 out of 43 patients with uterine cervical cancer (32.6%). The role of LT in cancer patients was discussed.

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Two forms of DNA polymerase α were separated from HeLa cells by DEAE-cellulose column chromatography and affinity chromatography using a DNA-cellulose column. They were eluted from a DEAE-cellulose column at 0.22M KCl (P-I) and 0.24M KCl (P-II). Upon cell fractionation between cytosol and nuclei, P-I was recovered from nuclei and P-II from cytosol. P-I was found to have a higher binding affinity to DNA than P-II by DNA-cellulose column chromatography.

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In more than 80% of tumor cells in the pericardiac effusion of a case of malignant B-cell lymphoma, Epstein-Barr virus-determined nuclear antigen (EBNA) was detected by the anticomplement immunofluorescence test. Moreover, herpestype virus particles, although few in number, were demonstrated in the nucleus of lymphoma cells by an electron microscope.
Tumor cells in the pericardiac effusion were seeded at 98% purity after centrifugation on Ficoll-Conray and, soon after plating, they proliferated continuously without any lag phase of growth or cell death. Therefore, the established cell line was regarded as of tumor cell origin and named Fujimaki-II cell after patient's name.
On the other hand, Fujimaki-I cells were established from the biopsied tumor in the same way. These two cell lines, B-lymphocyte in nature, had both EB virus-related antigens and herpes-type virus particles.
Heterotransplantation of cultured cells and tumor tissue obtained at autopsy into athymic nude mice was not successful. Transformation of cord blood lymphocytes by the virus released from Fujimaki-II cell also failed. This might be the first case of non-Burkitt type lymphoma in which the EB virus genome was directly detected in the tumor cells.

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Eleven syngeneic tumors were induced by 3-methylcholanthrene in DDD mice. Their antigenicity in vivo was examined by measuring growth inhibition of transplanted tumors in mice immunized by implanting tumors into their tail and then cutting off the tail. Four tumors with different antigenicity were selected from them for study. The growth-suppressive effect of an immunotherapeutic agent, PS-K, on the four tumors was examined. The growth of highly antigenic tumors in mice was inhibited by PS-K, but that of weakly antigenic tumors was not, and a close correlation was found between the antigenicity of tumors and the antitumor activity of PS-K against them. No similar correlation was found between their antigenicity and their sensitivity to a mitotic poison, 6-mercaptopurine. These results suggest that the antitumor effect of an immuno-potentiator depends greatly on the antigenicity of tumors.

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The distribution of 4-nitroquinoline 1-oxide (4-NQO) reductase, assumed to be closely related to the carcinogenesis of 4-NQO, was investigated in the mucosa of canine digestive tract, and its results indicated following points.
1) The activity of 4-NQO reductase was highest in the esophagus, next in the stomach, and remarkably low in the small and large intestines.
2) There is no significant difference in the 4-NQO reductase activity between the upper, middle, and lower portion of the esophagus, but its activity was higher in the female than in the male in its upper and middle portions.
3) Among the esophageal tissue, its activity was high only in the mucous epithelium and very low in all other layers.
4) Most of the enzymic activity in the esophageal mucosa existed in the cytosol fraction and activity of the microsome fraction was remarkably low. Even if NADPH or NADH was used as the hydrogen donor, its activity was not different in the cytosol fraction, but the former was a better hydrogen donor in the microsome fraction.
5) In the gastric mucosa, the enzymic activity was equally high in various portions of the corpus ventriculi; the greater and lesser curvatures, anterior and posterior parietes, and fundus. It was remarkably low only in the pyloric antrum.

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Male Wistar rats were given repeated injections of sterigmatocystin (stg) and related compunds into the peritoneal cavity once a week for 23 weeks. Only stg alone produced mesothelioma in 20 of 40 rats given a total dose of 20∼25mg, while no mesotheliomas were found in rats injected O-acetyl-stg (AcO-stg) and dihydro-stg. There were two histological types in the mesothelioma induced by stg; the epithelioid and mesenchymal types. There was little loose connective tissue between the epithelioid cell tumors, whereas abundant dense collagen fibers were found between the cell nests of the mesenchymal cell types of mesothelioma. The hepatocellular carcinomas were found in a rat treated with stg and in five animals given AcO-stg. The fact that mesotheliomas could be induced in high incidence by direct intraperitoneal application of stg suggests a potent carcinogenicity of stg. Although both dihydro-stg and stg formed fine needle-like crystals when injected into the peritoneal cavity, dihydro-stg could not produce any mesothelioma. Dihydro-stg, reduction product of stg, has no double bond in the terminal furan ring. Therefore, the carcinogenicity of stg may be related more closely with the presence of a double bond in the terminal furan ring of stg than the physical form in the peritoneal cavity. AcO-stg may easily be absorbed from the peritoneum, so that it could not persist in the peritoneum for sufficient duration for carcinogenesis.

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A quantitative analysis was performed on the extramedullary hematopoietic foci which appeared in the liver during the early stage of hepatocarcinogenesis with 3'-methyl-4-(dimethylamino) azobenzene in rats. The frequency in the appearance of the foci reached a maximum at around 3 weeks of azo-dye feeding. Since the liver in this period has been known to show deviation of various characteristics toward fetal liver, an intimate correlation of the appearance of the foci and fetal character of the liver was suggested.
Histologically the hematopoietic foci were always observed in the sinusoidal space of original hepatocytes adjacent to the oval cell proliferating area.
Ultrastructurally the cells of the foci were identified as erythroblasts and were found in the space of Disse as seen in the hematopoiesis in fetal liver.

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When CDF₁ mice inoculated intraperitoneally with leukemia P388 were treated intraperitoneally with mitomycin-C, of which dose was not so effective, on day 1 and given intraperitoneally polysaccharide preparation, Krestin, on day 2, 5, or 8 after tumor inoculation, best therapeutic results judged from 60-day survivors were obtained by combined use of mitomycin-C on day 1 and Krestin on day 5. Besides mitomycin-C, none of anticancer drugs such as cyclophosphamide, 5-fluorouracil, and 6-mercaptopurine was effective in this system. From these results, it was concluded that in this system, enhancement of antitumor effect of mitomycin-C by Krestin depends on the time of Krestin administration and that the combination effect is observed with mitomycin-C but not with cyclophosphamide, 5-fluorouracil, or 6-mercaptopurine.

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Cytotoxic activity of blood monocytes from 19 healthy individuals was examined in vitro against the cultured floating cells of human breast cancer origin. Target tumor cells inoculated on monocyte cultures and maintained for 5 days were reduced in number, compared with the control cultures of tumor cells alone. Increased cytotoxicity of the monocytes was noted after in vitro activation by heat-killed Corynebacterium liquefaciens (Propionibacterium acnes). C. liquefaciens of the same dose as used for monocyte activation had no effect on the tumor cell growth. These results may indicate that blood monocytes have an antitumor activity and it can be strengthened by treatment with C. liquefaciens. Antitumor activity of blood monocytes and macrophages was discussed on a concept of the mononuclear phagocyte system. A possible development of a new assay method for macrophage function as a parameter of host defence state against cancer will be expected.

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Dichloroplatinum(II) complexes of 1, 2-cyclohexanediamine isomers were reported to have high antitumor activity against leukemia L-1210 and P-388, but they were hardly soluble in water and, therefore, dibromo- and diiodoplatinum(II) complexes of 1, 2-cyclohexanediamine isomers were prepared. They were not soluble in water but showed high antitumor activity. When the chloride ions are replaced with sulfate ions, they become soluble and also they are antitumor active. Bismonobromoacetatoplatinum(II) complexes were synthesized and tested.
Mono- and bis-(D-glucuronato)platinum(II) complexes of 1, 2-cyclohexanediamine isomers were newly synthesized. All of them were freely soluble in water and showed high antitumor activity with low toxicity. The chelated mono(D-glucuronato)platinum(II) was found to be more effective than the monodentate bis(D-glucuronato)platinum(II) complexes.

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Intracellular localization of gallium-67 was investigated in Morris hepatoma-7316A and Shionogi mammary carcinoma-115 cells by the cell fractionation method 48hr after an intraperitoneal injection of the nuclide. When lysosomes were purified from both tumors by discontinuous sucrose density gradient centrifugation, they had a strikingly high relative specific activity of the nuclide. From these results it was confirmed that gallium-67 is concentrated most specifically in the lysosomes of both tumor cells, which consist chiefly of phagolysosomes and can engulf only limited amount of foreign materials such as Triton and gallium-67.

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The motility of four strains of rat ascites tumor cells in an agar medium was studied quantitatively by means of time lapse cinematography. The motility of these cells differed characteristically with each strain. Yoshida sarcoma and AH-66F cells showed active locomotion and partial movement markedly, while AH-130 and AH-100B cells made no locomotion and only a slight movement.

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Carcinogenicity of seven homologs of N-alkyl-N'-nitro-N-nitrosoguanidine (N-alkyl-NNG), namely, N-methyl-NNG, N-propyl-NNG, N-butyl-NNG, N-isobutyl-NNG, N-pentyl-NNG, and N-hexyl-NNG, was examined by injecting the compounds subcutaneously into rats. All of them except N-isobutyl-NNG and N-hexyl-NNG were carcinogenic, but N-ethyl-NNG followed by N-methyl-NNG were stronger carcinogens than those with longer alkyl chains.

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The antiestrogen, Mepitiostane, an oral derivative of Epitiostanol (Thiodrol), was administered to patients with operable breast cancer as an endocrine adjuvant therapy. The estrogen receptor (ER) was measured in most cases. Patients with ER (+) tumors showed lower recurrence rate than the patients with ER (-) tumors.

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Newly synthesized 5-fluorouracil (5-FU) O-β-D-glucuronide and 5-FU N-glucuronide were tested for their antiumor effect using DDD mouse mammary carcinoma. Only the former was effective if glucose treatment of the host was combined. Toxicity of both compounds on mice was proved to be very low (ca. 5000mg/kg, iv).

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4-Nitrosoquinoline 1-oxide (4-NOQO) was found to be converted reductively into the same free radical as that produce oxidatively from 4-hydroxyaminoquinoline 1-oxide (4-HAQO). By the fluorometric method 4-NOQO was proved to bind covalently with polyguanylic acid more effectively than 4-HAQO.

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Adriamycin induced endognous murine leukemia virus from AKR-2B mouse cell line. The optimum concentration for the virus induction was 0.1μg/ml. at this concentration, the number of viable cells was about 12% relative to the control.

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Activation of cytotoxic macrophages with lymphokine, a macrophage activating factor, was prevented by pretreatment of macrophages with either aggregated IgG or Fc fraction of IgG, but neither with heated BSA, monomeric IgG, nor F(ab')₂ suggesting that the inhibition was mediated by Fc receptor on macrophages.

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