bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Intraperitoneal Injection of Mitomycin C Adsorbed on Activated Carbon Particles on Induction of Cytolytic Peritoneal Macrophages of Mice
Authors: Sakita Masakazu , Iwamoto Akihiko , Hata Kouki , Hagiwara Akeo , Takahashi Toshio
Published: August 2005

The effects of the intraperitoneal injection of mitomycin C adsorbed onto activated carbon particles (MMC‐CH) were assessed on the host immune status and the induction of cytolytic peritoneal macrophages (PM) in mice in comparison with an aqueous MMC solution (MMC‐AQ). PM from inbred C57BL/6 mice and syngeneic B16 melanoma cells were used as the effector and target cells for the PM‐mediated cytotoxicity assay. After a single injection of MMC‐AQ (at the dose of 1/2 LD50), the weights of thymus and spleen and the 3H‐thymidine uptake of spleen cells stimulated by concanavalin A were markedly decreased compared to those following injection of MMC‐CH containing the same dose of MMC. A significant difference was found in the chronological changes of peritoneal exudate cell (PEC) numbers and PM cytolytic activity between the MMC‐AQ‐ and MMC‐CH‐treated mice. One day after MMC‐AQ injection, the PEC numbers were decreased markedly. However, they increased ten‐fold after MMC‐CH injection and the PM cytolytic activity was significantly higher after MMC‐CH injection than after MMC‐AQ injection during the first 3 days. The present results suggest the possible superiority of MMC‐coated charcoal over free MMC both because of a lessening of the effects on host immunity due to prolonged slow drug release and because of the increase in cytolytic macrophages that was induced.
Pharmacokinetic Modulation of Plasma 5‐Fluorouracil Concentrations to Potentiate the Antitumor Activity of Continuous Venous Infusion of 5‐Fluorouracil
Authors: Fujii Setsuro , Fukushima Masakazu , Shimamoto Yuji , Shirasaka Tetsuhiko
Published: August 2005

Methods for pharmacokinetic modulation of the plasma 5‐fluorouracil (5‐FU) level to increase antitumor activity during continuous venous infusion (CVI) of low doses of 5‐FU were examined in Yoshida sarcoma‐bearing rats. These methods were additional infusion of 5‐FU for a short period (4 h) or oral administration of LIFT or Tegafur during long‐term CVI of 5‐FU that alone gave a plasma 5‐FU level of about 50 ng/ml. The antitumor effect on Yoshida sarcoma was markedly potentiated when an additive dose of 5‐FU combined with 3‐cyano‐2,6‐dihydroxypyridine (CNDP), a potent inhibitor of 5‐FU degradation, giving a plasma level of about 500 ng/ml, was infused for 4 h. A similar increase in the antitumor effect was observed with oral administration of a conventional dose of UFT during CVI of 5‐FU without CNDP, giving a plasma level of 30 to 60 ng/ml. These results suggest that the antitumor effect of CVI of 5‐FU can be potentiated by pharmacokinetic modulation of the 5‐FU concentration in the blood.
Augmentation of Human Cytotoxic T Lymphocytes against Autologous Tumor by a Factor Released from Human Monocytic Leukemia Cell Line
Authors: Maeda Nagamasa , Hamasato Shinji , Miyazawa Hitoshi , Takata Masaru , Yamamoto Hiroshi , Fujimoto Shigeyoshi
Published: August 2005

A human acute monocytic leukemia cell line, THP‐1, releases a factor which activates human cytotoxic (killer) T lymphocytes (CTL) against autologous tumor in vitro. The factor, named cytotoxic (killer) T cell activating factor (KAF), is an acidic protein of 70,000 to 100,000 dalton molecular size. Peripheral blood leukocytes from two patients, bearing epithelioid sarcoma or malignant schwannoma, were cultured for 7 days with individual autologous tumor to induce CTL directed to the corresponding tumor. Monocyte‐depleted peripheral leukocytes generated lesser CTL activity than the monocyte‐containing leukocyte population. However, the KAF was able to replace the monocyte function. The KAF acted at the CTL generation phase as well as the effector phase. The KAF‐activated killer cells possessed CD4‐8+ surface phenotype. The CTL killed autologous tumor or other unrelated tumor cell lines only when they shared some of the HLA class I antigens. It was also demonstrated that the KAF does not activate killer cells without proper antigenic stimuli, because the KAF‐augmented CTL possess specificity against autologous tumor or other HLA‐A or ‐B matched tumor cell lines. The therapeutic applicability of human KAF for anti‐tumor CTL therapy against autologous tumor is discussed.
Comparative Antitumor Activity of 5‐Fluorouracil and 5′‐Deoxy‐5‐fluorouridine in Combination with Radiation Therapy in Mice Bearing Colon 26 Adcnocarcinoma
Authors: Ishikawa Tohru , Tanaka Yutaka , Ishitsuka Hideo , Ohkawa Tomohiko
Published: August 2005

The present study compared the antitumor activities of chemotherapy with 5‐fluorouracil (5‐FU) and with its prodrug 5′‐deoxy‐5‐fluorouridine (5′‐DFUR) in combination with radiotherapy on a solid colon 26 adenocarcinoma in the mouse. A single administration of 5′‐DFUR immediately after local irradiation on day 10 after tumor inoculation produced more than additive antitumor effects, while only an additive effect was observed in the combined treatment with 5‐FU and radiation. This over‐additive effect of 5′‐DFUR was more obvious in a fractionated‐dose treatment schedule, where the same combined modality treatment was given three times on days 6, 10 and 14 after inoculation of the tumor cells. 5′‐DFUR enhanced the radiation effects on the tumor in terms of the delay in tumor growth as well as the increase in the survival time. 5‐FU produced only a marginal additive antitumor effect. Furthermore, radiation damage to normal tissues (skin damage by local irradiation and bone marrow and spleen damage by whole‐body irradiation) was not enhanced by 5′‐DFUR, though radiation damage to the thymus was additive. On the other hand, 5‐FU produced toxic effects that were additive for all normal tissues tested. Thus, at doses that were the most effective against tumors, relative therapeutic gain factors (the ratio of the effect on tumors to that on the bone marrow) of 5′‐DFUR and 5‐FU were 1.24 and 0.49, respectively. These results suggest that 5′‐DFUR will have a greater potential than 5‐FU in combined modality treatment of cancer patients.
Frequency and Types of Point Mutation at the 12th Codon of the c‐Ki‐ras Gene Found in Pancreatic Cancers from Japanese Patients
Authors: Mariyama Mariko , Kishi Kiyozo , Nakamura Kozo , Obata Hiroshi , Nishimura Susumu
Published: August 2005

Point mutations at the 12th codon of c‐Ki‐ras in pancreatic cancer from Japanese patients were examined using the polymerase chain reaction, followed by cloning of the amplified gene fragments in pTZ phagemid and nucleotide sequence determination. The frequency of the point mutations found in the tumors was quite high (75%). The mutation most frequently detected was a G→A transition at the second position of codon 12 (GGT→GAT), but other types of mutations such as GGT→GTT and GGT→CGT were also found. In one case, silent mutation of GGT to GGC was detected in addition to the frequent mutation of GGT to GAT, These observations suggest that the 12th codon of pancreatic c‐Ki‐ras is highly mutatable.
The Roles of CD8+ and CD4+ Cells in Tumor Rejection
Authors: Udono Heiichiro , Mieno Masahiro , Shiku Hiroshi , Nakayama Eiichi
Published: August 2005

In vivo administrations of anti‐Lyt‐2.2 (CDS) mAb and anti‐L3T4 (CD4) mAb selectively eliminated CD8+ cells amd CD4+ cells, respectively. The relative potencies of CD8+ cells and CD4+ cells and their roles in primary tumor rejections were studied by investigating the effects of these mAbs on tumor growth. CD8+ cells were themselves fully capable of mediating rejection in 5 different tumor rejection systems: two radiation leukemia virus (RadLV)‐induced leukemias, B6RV2 and BALBRVD, a radiation‐induced leukemia BALBRL♂1, and a plasmacytoma BALBMOPC‐70A in CB6F1 mice, and a Friend virus‐induced leukemia B6FBL‐3 in B6 mice. On the other hand, CD4+ cells were capable of resisting tumor growth of B6FBL‐3, but not of the other four tumors. Furthermore, for efficient rejection of CB6F1UV+˚l sarcoma by CB6F1 mice, synergy of CDS+ and CD4+ cells was necessary. Blocking of UV+˚ 1 rejection was abrogated by delayed administration of anti‐L3T4 (CD4) mAb but not anti‐Lyt‐2.2 (CDS) mAb, indicating the involvement of CD4+ cells in only the initial phase of rejection.
Antitumor Activity and Pharmacology of 1‐β‐D‐Arabinofuranosylcytosine‐5′‐stearylphosphate: An Orally Active Derivative of 1‐β‐D‐Arabinofuranosylcytosine
Authors: Kodama Kenjiro , Morozumi Manami , Saitoh Ken‐ichi , Kuninaka Akira , Yoshino Hiroshi , Saneyoshi Mineo
Published: August 2005

The antitumor activity of l‐β‐D‐arabinofuranosylcytosine‐5′‐alkylphosphates (CnPCAs) against L1210 leukemia in mice after oral administration was demonstrated. The optimum length of the alkyl group on the phosphate moiety of CnPCA for exhibiting a high antitumor activity was found to be between tetradecyl (C14) and tricosyl (C23). The most active alkyl derivative in this system was found to be l‐β‐D‐arabinofuranosylcytosine‐5′‐stearylphosphate (C18PCA). The optimum and minimum effective doses of C18PCA were 100 and 6,25 mg/kg/day (q1d, day 1 to day 5), respectively. The maximum T/C% of C18PCA was approximately 220. The antitumor activity of C18PCA was not greatly dependent on the treatment schedule and route. Plasma concentration of 1‐β‐D‐arabinofurano‐sylcytosine (ara‐C) remained in the range of 0.4 to 0.75 μmol/ml for 24 h after oral administration of 100 mgAg (170 μmol/kg) of C18PCA. These results indicate that C18PCA administered per orally is absorbed intact through the gastrointestinal tract and ara‐C is released for a long period of time. C18PCA is regarded as an orally active depot form of ara‐C
Increased Expression of ras Genes in Non‐Hodgkin's Lymphomas Is not Associated with Oncogenic Activation of Those Genes by Point Mutation
Authors: Hamatani Kiyohiro , Nagata Yasuhiko , Abe Masumi , Abe Kuniko , Toki Hironobu , Ikeda Shuichi , Nakayama Eiichi , Shiku Hiroshi
Published: August 2005

Twenty‐three cases of non‐Hodgkin's lymphoma (NHL) were analyzed for expression of ras genes by in situ hybridization utilizing biotinylated DNA probes. Increased expression of Ki‐ras, Ha‐ras and N‐ras genes was observed in 12 cases, 6 cases and 1 case of NHL, respectively. Genomic DNA extracted from these 23 cases of NHL was region‐specifically amplified by means of polymerase chain reaction to examine the presence of point mutations at the 12th, 13th and 61st codons of Ki‐, Ha‐ and N‐ras genes. Dot hybridization assays with appropriate oligonucleotide probes showed no evidence of point mutation in any case of NHL examined. These results indicate that increased expression of ras genes in NHL is not associated with ras gene activation by point mutation.
T‐Cell Phenotype Is Associated with Decreased Survival in Non‐Hodgkin's Lymphoma
Authors: Shimizu Kazuyuki , Hamajima Nobuyuki , Ohnishi Kazunori , Hara Kazuo , Kunii Akira
Published: August 2005

This study was undertaken to determine which if any presentment factors are statistically significant determinants of the clinical outcome in patients with non‐Hodgkin's lymphoma. The pretreatment factors in 20 patients with T‐cell lymphoma, including two patients with adult T‐cell leukemia/ lymphoma (ATLL), and 28 patients with B‐cell lymphoma were evaluated. In a stepwise logistic regression analysis, a T‐cell phenotype in addition to high grade histology and pleural involvement demonstrated a statistically significant correlation with decreased response rate, when the analysis did not include patients with ATLL. Analysis by means of the Cox proportional hazards model disclosed that the T‐cell phenotype retained a statistically significant correlation with survival after adjustments for other prognostic factors, whether the study included the patients with ATLL or not. The decreased response rate and survival of Japanese patients with non‐Hodgkin's lymphoma in comparison with those reported in Western countries seem to be due to increased intrusion of T‐cell lymphomas. To permit a reliable comparison of reports on new chemotherapeutic regimens from different institutions, the tumor phenotype must be determined in the population studied
Combination Effects of Forty Carcinogens Administered at Low Doses to Male Rats
Authors: Takayama Shozo , Hasegawa Hirokazu , Ohgaki Hiroko
Published: August 2005

An investigation was conducted to determine whether a mixture of low doses of forty carcinogens that target different organs, including the liver, intestine, thyroid, urinary bladder, and skin, is effective for tumor induction in F344/DuCrj rats. The dose of each carcinogen in the diet was 1/50 of the TD50 value, treatment being continued for 102 weeks. Significant numbers of neoplastic nodules of the liver and follicular cell tumors of the thyroid developed in the animals exposed to the carcinogen mixture, although the question of whether the observed carcinogenic effects were synergistic or additive could not be answered. The results serve to evaluate carcinogenic risk in the search for causes of human cancer.
Epidermal Growth Factor Induces the Expression of Its Receptor Gene in Human Gastric Carcinoma Cell Line TMK‐1
Authors: Yoshida Kazuhiro , Takanashi Atsushi , Kyo Eikai , Ito Masanori , Ito Hisao , Niimoto Minoru , Hattori Takao , Tahara Eiichi
Published: August 2005

To ascertain the possible autocrine pathway in the growth promotion of gastric carcinomas, a study was made on the effects of exogenous human epidermal growth factor (hEGF) on the expression of mRNA for EGF, transforming growth factor‐α (TGF‐α), EGF receptor, FOS and MYC genes by TMK‐1 cells. Exogenous hEGF increased FOS and MYC mRNA levels 30 min and 1 h after the treatment, respectively. TMK‐1 cells accumulated the mRNA for EGF receptor about 7‐ to 8‐fold by 3 h after treatment Expressions of mRNA for EGF and TGF‐α genes were detected, but the amounts of the mRNA of these genes in TMK‐1 cells were not altered after the treatment.
Detection of HTLV‐I Genome in Seronegative Infants Born to HTLV‐I Seropositive Mothers by Polymerase Chain Reaction
Authors: Saito Shigeru , Ando Yoshiya , Furuki Kazuhiko , Kakimoto Kazuhiro , Tanigawa Takuo , Moriyama Ikuko , Ichijo Motohiko , Nakamura Masataka , Ohtani Kiyoshi , Sugamura Kazuo
Published: August 2005

We applied the polymerase chain reaction (PCR) method to detect gag, env and pX sequences of human T cell leukemia virus type I (HTLV‐I) provirus in peripheral blood lymphocytes of seronega‐tive infants born to HTLV‐I seropositive mothers. Out of 22, five subjects were found to contain the HTLV‐I provirus genome. Two of the five cases were judged to be negative for not only anti‐HTLV‐I antibodies but also the viral antigens on cultivated lymphocytes by the conventional antibody/antigen detection methods. These results indicate that PCR is of great use as a simple and highly sensitive method detect HTLV‐I infection.
Expression of Type II and Type XI Collagens in Canine Mammary Mixed Tumors and Demonstration of Collagen Production by Tumor Cells in Collagen Gel Culture
Authors: Aral Katsuhiko , Uehara Kohkichi , Nagai Yutaka
Published: August 2005

The development of cartilaginous collagen types, II and XI, in canine mammary mixed tumors was studied biochemically and immunohlstochemically. In mixed tumor, an alcian blue‐positive myxomatous region appeared in the stroma, where round‐shaped proliferating myoepithelial cells were scattered. Type II collagen was distributed in metaplastic cartilage matrix, while type XI was located only in the pericellular region, where proliferating cells were positively stained with anti‐actin and anti‐keratin antibodies. The accumulation of collagen types II and XI in the tumor mass was confirmed by sodium do decyl sulfate‐polyacrylamide gel electrophoresis followed by immunoblotting of the extract of the lesion using type‐specific antibodies to collagen types II and XI. Tumor cells isolated from metaplastic tumor mass expressed both collagen types II and XI and myoepithelial types of cytoskeleton in gel culture, in which an alcian blue‐positive substance became detectable in the pericellular region on day 3 and type II and type XI collagens on day 5. This may be a useful model for studying chondrocyte‐type gene expression during tumorigenesis

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Herbimycin A Suppresses the Reduction of Gap‐junctional Intercellular Communication Induced by Tumor‐promoting Phorbol Ester in 3T3‐L1 Cells
Authors: Sasaki Yasuto , Shiba Yoshiki , Hirono Chikara , Kanno Yoshinobu , Takeuchi Tomio , Umezawa Kazuo
Published: August 2005

We have examined the suppressive effect of herbimycin A on the reduction of gap‐junctional Intercellular communication that is induced by a tumor‐promoting phorbol ester in 3T3‐L1 cells. Most cells in growth arrest participated in dye‐coupling, as evaluated by the transfer between cells of a fluorescent dye (Lucifer Yellow CH). Treatment of cells with 0.25 μg/ml herbimycin A slightly enhanced the dye‐coupling. This enhancement required treatment for periods as long as 24 h. Addition of 100 ng/ml 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA) caused a rapid reduction of dye‐coupling. However, addition of TPA did not suppress dye‐coupling in cells pretreated for more than 24 h with herbimycin A. Pretreatment of cells for less than 6 h with herbimycin A did not suppress the TPA‐induced reduction of dye‐coupling. These results suggest that herbimycin A suppresses the reduction of gap‐junctional intercellular communication that is induced by TPA through enhancement of the ability of the cells to participate in gap‐junctional intercellular communication
Binding Properties and Proliferative Effects of Human Recombinant Granulocyte‐Macrophage Colony‐stimulating Factor in Primary Leukemia and Lymphoma
Authors: Piao Yun‐Feng , Chiba Shigeru , Tojo Arinobu , Urabe Akio , Nara Nobuo , Miyazono Kohei , Takaku Fumimaro
Published: August 2005

Binding of radiolabeled human granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) was studied with blast cells front eight patients with acute myeloblastic leukemia (AML), and neoplastic lymphoid cells from one patient with acute lymphoblastic leukemia (ALL), two patients with chronic lymphocytic leukemia (CLL) and one patient with undiagnosed B cell neoplasia. In all AML cases studied, Scatchard graphs of the direct binding data were curvilinear, and were best fitted by curves derived from a two‐binding‐site model; one site with high affinity (Kd1= 12–71 pM; 174–602 sites/cell) and the other with low affinity (Kd2= 0.5–2,7 nM; 1137–6020 sites/cell). A cross‐linking study on blast cells from one AML patient demonstrated specific bands which were similar to those reported for peripheral blood neutrophils. Furthermore, blast colony assays for the same preparations showed remarkable proliferative response to GM‐CSF in the concentration range from 0.3 nM to 7.0 nM (ED50 >0.7 nM). This concentration range is approximately one order of magnitude higher than that which is effective for colony formation from normal bone marrow progenitors (ED50= 0.l nM). No significant correlation could be observed between the responsiveness of blast progenitors to GM‐CSF, and the numbers or affinities of GM‐CSF binding sites demonstrated on blast cells. In studies with neoplastic lymphoid cells from four patients, 125I‐GM‐CSF also specifically bound in two cases, while response to GM‐CSF was not observed in these cases. These results indicate that the expression of GM‐CSF receptor is not restricted to the GM‐CSF‐responsive AML blast cells, but can be observed in other AML blast cells and even in neoplastic lymphoid cells.
Combination Therapy of Colon Carcinoma 26 in Mice with Recombinant Human Interleukin‐2 and Interferon‐α A/D: Occurrence of Large Granular Cells in the Tumor
Authors: Sakura Yasufumi , Ootsu Koichiro , Shino Akio
Published: August 2005

The antitumor effects of recombinant human interleukin‐2 (rIL‐2), in combination with recombinant human interferon‐α A/D hybrid (rIFN‐α A/D) on colon carcinoma 26 (colon 26) in mice were examined histologically. Colon 26 was transplanted subcutaneously into female BALB/c mice on day 0. The mice bearing the tumor received intramuscular injection of rIL‐2, rIFN‐α A/D or the combination of rIL‐2 and rIFN‐α A/D for 2–10 consecutive days starting on day 7. Mice were killed on days 9, 13, 17 and 21. After day 13, growth of the tumor was significantly suppressed in the mice treated with rIL‐2 or rIFN‐α A/D alone and was stopped in the mice treated with rIL‐2 in combination with rIFN‐α A/D. Histologically, tumor necrosis developed in all treated groups, though the degree was the most severe in the group receiving combination treatment. Many large cells (about 15 30 μm in diameter) infiltrated into the tumor, and they had Thy‐1 surface antigen and many periodic acid‐Schiff‐positive round grannies in the cytoplasm. The incidence of these large granular cells was correlated well with the reduction in tumor weight. The ultrastructural features of the large granular cells were very similar to those of murine large granular lymphocyte‐like cells maintained in vitro in an IL‐2‐containing medium. The present large granular cells appear to be a kind of activated lymphoid cells.
A Murine Monoclonal Antibody that Recognizes an Extracellular Domain of the Human c‐erbB‐2 Protooncogene Product
Authors: Masuko Takashi , Sugahara Kunio , Kozono Mitsuko , Otsuki Seiko , Akiyama Tetsu , Yamamoto Tadashi , Toyoshima Kumao , Hashimoto Yoshiyuki
Published: August 2005

A marine IgM monoclonal antibody, designated SV2–61, was generated against human c‐erbB‐2 gene‐transfected NIH‐3T3 (SV11) cells. SV2–61 defined a 185‐kDa molecule present on the surface of SV11 cells, another line of c‐erbB‐2 gene‐transfected NIH‐3T3 (A4–15) cells, and MKN‐7 human gastric cancer cell line carrying an amplified human c‐erbB‐2 gene. The SV2–61‐defined antigen was found to show protein kinase activity in vitro. The SV2–61 was reactive with human c‐erbB‐2 gene‐transfected NIH‐3T3 cell lines but not with transfectants carrying c‐erbB‐2 gene mutants which lack a coding region for the extracellular domain. It was reactive with a portion of human epithelial cell lines hut not with native NIH‐3T3, TGF‐a‐coding gene‐, activated c‐rafgene‐ or Ha‐ras gene‐transfected NIH‐3T3 cells, or non‐epithelial human cells. These results indicate that the SV2–61 is an antibody which recognizes an extracellular domain of the c‐erbB‐2 gene product, 185‐kDa protein.
Modification of Cellular Membrane Functions by Pendolmycin
Authors: Umezawa Kazuo , Imoto Masaya , Yamashita Takashi , Sawa Tsutomu , Takeuchi Tomio
Published: August 2005

Pendolmycin is a new indole alkaloid isolated from Nocardiopsis as an inhibitor of phosphatidyl‐inositol turnover. Structurally, pendolmycin is similar to teleocidin B and lyngbyatoxin A. Pendolmycin, teleocidin B and 12‐O‐tetradecanoylphorbol‐13‐acetate all inhibited phosphatidylinositol turnover in A431 cells. Pendolmycin inhibited binding of epidermal growth factor, activated ara‐chidonic acid release and hexose transport, and inhibited binding of phorbol‐12,13‐tlibutylate in C3H10T1/2 cells. Thus, pendolmycin was as potent as teleocidin B and phorbol ester tumor promoters in modification of cell membrane functions, while having the simplest structure among them.
Biochemical and Immunological Characterization of Murine Leukemia Viruses that Are Paralysis‐inducing in Rats
Authors: Shibamura Sei‐Ichi , Kai Kazushige , Akatsuka Toshitaka , Odaka Takeshi
Published: August 2005

The molecular size and pi of the viral structural proteins of four PVC viruses (PVC111, PVC211, PVC321 and PVC441) were compared by single or two‐dimensional polyacrylamide gel electrophoresis. PVC111 had slightly larger plSE and gPr85 molecules (about 0.5 kilodalton) than did the other PVC viruses. On the other hand, the virion structural proteins p30, p15, p12E and p12 from all the viruses had the same molecular sizes and pis. The gp70s and plOs from all the viruses showed the same molecular sizes. A monoclonal antibody to gp70 of PVC321 virus recognized the gp70s of all PVC viruses, but not the gp70s of four clones of the wild mouse ecotropic viruses, Friend murine leukemia viruses (F‐MuLV), AKR ecotropic MuLV, dual‐tropic F‐MuLV or NZB endogenous xenotropic MuLV, revealing that these four PVC viruses are homologous with each other, but distinct from the known mouse retrovirnses.
Cultured Skin Fibroblasts from Patients with Porokeratosis Are Hypersensitive to the Lethal Effects of X‐radiation
Authors: Otsuka Fujio , Watanabe Ryohji , Moro Akiko , Ohkochi Hitoshi , Ishibashi Yasumasa
Published: August 2005

Porokeratosis is an autosomal dominant inherited skin disorder. The lesions are characterized by localized abnormal keratinization and may develop into malignant tumors. To determine the cellular basis of the cancer susceptibility associated with this skin condition, we examined the colony‐forming ability of X‐ray or ultraviolet (UV) light irradiated, cultured fibroblasts derived from porokeratosis patients' normal‐appearing skin. Four fibroblast strains derived from four porokeratosis patients' skin were significantly hypersensitive to the lethal effects of X‐radiation. However, they all showed a similar sensitivity to strains from normal donors to 254 nm UV light. The hypersensitivity to X‐ray radiation in cultured skin fibroblasts from porokeratosis patients suggests an inherent instability of cellular DNA and may probably be associated with the cancer‐prone nature of this skin condition.
Lysis of Human Leukemic Cells by Monocyte‐derived Macrophages Activated with Interferon‐γ and Interleukin‐2
Authors: Kakita Tokio , Sasada Masataka , Moriguchi Toshinori , Nishimura Toshiro , Yamamoto Kohkichi , Uchino Haruto
Published: August 2005

Cytolysis of leukemic cells by peripheral blood‐derived macrophages was examined by means of an in vitro111In release assay. Monocytes prepared on culture dishes lyse YK‐M2. However, when monocytes were cultured in vitro and transformed into macrophages, they lost most of their lytic activity. The addition of human recombinant interleukin‐2 (rIL‐2) on day 5 in culture enhanced the lytic activity significantly. Similarly, treatment of macrophages with human recombinant interferon gamma (rIFN‐γ) promoted the lysis of YK‐M2 and K‐562, although the extent of lysis was smaller than that by rIL‐2. Macrophages activated with rIL‐2 and rIFN‐γ also Iysed human leukemic cells. Activated macrophages lysed leukemic cells of acute myelocytic leukemia more than acute lymphocytic leukemia cells. Macrophages derived from the peripheral blood of patients with leukemia were examined for their lytic activity against YK‐M2. The patient's macrophages lysed more YK‐M2 than did control macrophages when they were activated with rIL‐2 and rIFN‐γ. The macrophages of two patients also demonstrated autologous leukemic cell lysis.
Inhibition of Intracellular Bleomycin Hydrolase Activity by E‐64 Leads to the Potentiation of the Cytotoxicity of Peplomycin against Chinese Hamster Lung Cells
Authors: Nishimura Chiaki , Nishimura Toshio , Tanaka Nobuo , Yamaguchi Hideyo , Suzuki Hideo
Published: August 2005

N‐(N‐(L‐3‐frans‐carboxyoxiran‐2‐carbonyl)‐L‐leucyl)‐agmatine (E‐64), a thiol protease inhibitor, potentiated the cytotoxicity of peplomycin against the Chinese hamster lung (V79) cell. After the treatment of the cells with E‐64 (50 μg/ml) for 12 h, bleomycin hydrolasc activity of the cells was almost completely inhibited. V79 cells treated with [3H]peplomycin for 24 h in the presence of E‐64 (50 μg/ml) accumulated twice as much [3H]peplomycin and five times less [3H]desamidopeplomycin compared with V79 cells treated in the absence of E‐64. These results suggest that E‐64 increases the sensitivity of V79 cells to peplomycin probably by inhibiting the intracellular bleomycin hydrolasc activity.
Antitumor Activities of a Novel 9‐Aminoanthracycline (SM‐5887) against Mouse Experimental Tumors and Human Tumor Xenografts
Authors: Morisada Shinya , Yanagi Yoshikazu , Noguchi Toshihiro , Kashiwazaki Yasuo , Fukui Masaru
Published: August 2005

The antitumor effects of SM‐5887, a totally synthetic 9‐aminoanthracycline derivative, were evaluated in six murinc experimental tumor systems (P38S, Ehrlich carcinoma, sarcoma 180, Lewis lung carcinoma, B16 melanoma and colon 38) and nine human tumor‐nude mouse systems (one breast cancer, two lung cancers and six gastric cancers). Characteristically SM‐5887 showed excellent antitumor activities, superior to adriamycin (ADR), against human tumor xenografts, although its activities against murine experimental tumors were almost equal to those of ADR. When the human tumors were implanted sc in female athymic mice (BALB/c, nu/nu) and their volume reached 100‐ 300 mm3, SM‐5887 and ADR were injected iv. All nine human tumors tested showed statistically significant responses to SM‐5887, and 7 of them were strongly suppressed in their growth by SM‐5887 so that minimum T/C values were less than 30% at the maximum tolerated dose (MTD, 25 mg/kg) with a single iv injection. Compared with ADR, SM‐5887 was statistically more effective in five tumors (one breast, one lung and three gastric), equal in two tumors (two gastric), and less potent in two tumors (one lung and one gastric). In addition, the 10‐day‐interval repeated iv treatments with SM‐5887 at the MTD (25 mg/kg) resulted in remarkably potent antitumor effects (including complete regression) against human gastric cancer, 4‐1ST, implanted in nude mice without enhancement of toxic effects, SM‐5887 was also effective against ip‐inoculated P388 by oral administration as well as iv injection.
Effect of Adriamycin on the Activities of Superoxide Dismutase, Glutathione Peroxidase and Catalase in Tissues of Mice
Authors: Sazuka Yasuyuki , Tanizawa Hisayuki , Takino Yoshio
Published: August 2005

The increment of lipid peroxide in the hearts of mice treated with adriamycin (ADR) was examined in relation to the decrease in the activities of Superoxide dismutase (SOD), glutathione peroxidase (GSHpx) and catalase. The natural activities of these enzymes in mouse heart are lower than those in the liver. The biggest decrease in enzyme activity observed in the heart after ADR administration was that of GSHpx. Therefore, the increment of lipid peroxide was attributable to the decrease in the activities of these enzymes, especially GSHpx. Subsequently, the effects of antioxidants on the decreases in activities of SOD, GSHpx and catalase in the hearts of mice treated with ADR were examined. However, the decrease in the activities of the enzymes were not accompanied with any increment of lipid peroxide. This result suggests that active oxygen radicals produced by ADR through the agent's redox cycling have no effect on the activities of these enzymes. Therefore, it appears that the decrease in the activities of these enzymes induced by ADR in the mouse results from inhibition of enzyme protein biosynthesis.
The Biphasic Change of Cytosolic Acetyl‐CoA Hydrolase in Rat Liver during 3′‐ Methyl‐4‐dimethylaminoazobenzene Hepatocarcinogenesis
Authors: Ebisuno Shoji , Isohashi Fumihide , Nakanishi Yoko , Higashi Taneaki , Sakamoto Yukiya
Published: August 2005

When Donryu male albino rats were given diet containing 0.06% 3′–methyl‐4‐dimethyl‐ aminoazobenzene (3′–Me‐DAB) for 20 weeks, the activity of cytosolic acetyl‐CoA hydrolase in their livers decreased to about one‐third the initial level in week 2, returned to the control level in week 7, and then decreased again to about one‐tenth of the control in week 20. These changes in enzyme activity were parallel with changes in the amount of enzyme protein determined by ELISA. In 3′–Me‐DAB‐resistant rats, however, the enzyme activity and enzyme protein remained within the normal range during administration of 3′–Me‐DAB‐containing diet for 20 weeks and no tumors were detectable macroscopically. Interestingly, the biphasic change in this enzyme activity was inversely associated with the well known change of γ‐glutamyltranspeptidase activity during azo‐dye‐induced hepatocarcinogenesis.
Differential Activation of Lymphokine‐activated Killer Cells with Different Surface Phenotypes by Cultivation with Recombinant Interleukin 2 or T‐cell Growth Factor in Gastric Cancer Patients
Authors: Koyama Shohei , Ebihara Tsugio , Fukao Katashi , Osuga Toshiaki
Published: August 2005

Fourteen days' culture of human peripheral blood lymphocytes (PBL) with recombinant interleukin 2 (rIL 2) or T cell growth factor (TCGF) results in the generation of lymphoklne‐activated killer (LAK) effector cells which have the unique property of lysing natural killer (NK)‐resistant human tumor cells, Daudi, as well as NK‐sensitive, K562 cells. LAK cells were generated from both normal and gastric cancer patients' PBL. However, LAK cell activities induced by rIL 2 or TCGF decreased with the progress of the tumor growth. In addition, TCGF‐induced LAK cell activities were found to be lower than the rIL 2‐indnced LAK cell activities. Different mechanisms may be involved in the decreases of the rIL 2‐induced and TCGF‐induced LAK cell activities. This study further demonstrates that the cell types involved are also heterogeneous, as determined by phenotypic characteristics. The LAK‐effector cell type was analyzed by two‐color flow cytometry. RIL 2‐induced LAK cells showed increased proportions of CD4±Leu 8‐ and Leu 7±CD16‐, and a decreased proportion of CD8±CD11‐ cells, which are believed to be associated with killer T cell functions. In contrast, TCGF‐induced LAK cells revealed significantly increased proportions of CD8±CD11‐and CD4±Leu 8‐ cells, and a decreased proportion of Leu 7±CD16‐ cells. Thus, LAK cells with different surface phenotypes were induced by the cultivations with rIL 2 and with TCGF.
Transformation of NIH3T3 Cells with Synthetic c‐Ha‐ras Genes
Authors: Kamiya Hiroyuki , Miura Kazunobu , Ohtomo Noriko , Koda Toshiaki , Kakinuma Mitsuaki , Nishimura Susumu , Ohtsuka Eiko
Published: August 2005

Synthetic human c‐Ha‐ras genes in which amino acid codons were altered to those which are frequently used in highly expressed Escherichia coli genes were ligated to the 3′‐end of Rous sarcoma virus long terminal repeat. When NIH3T3 cells were transfected with the plasmids having those genes with valine at codon 12, leucine at codon 61 or arginine at codon 61, transformants were efficiently produced. These results indicated that the synthetic c‐Ha‐ras genes are expressed in a mammalian system even though their codon usage is altered to correspond with that of E. colt. This expression vector system should he useful for studies on the structure‐function relationships of c‐Ha‐ras, since the synthetic gene can be easily modified to have multiple base alterations, and can also be used simultaneously for the production of large amounts of p21 in E. coli for biochemical and biophysical studies.
Lung Cancer Cells Often Express High Levels of Protein Kinase C Activity
Authors: Hirai Masamichi , Gamou Shinobu , Kobayashi Makoto , Shimizu Nobuyoshi
Published: August 2005

We analyzed protein kinase C (PKC) activity in twenty‐two tumor cell lines derived from lung, pancreas, stomach, tongue and vulva, and found that lung cancer cells often (9 out of 13) exhibit significantly higher PKC activity than other types of cancer cells. The PKC in these lung cancer cells was separated into one major and one minor peaks by a Mono Q column chromatography. The PKC in the major peak had an absolute requirement for Ca2±, phosphatidylserine and 12‐O‐tetradeca‐noylphorbol‐13‐acetate (TPA), as expected. However, the PKC in the minor peak did not require TPA for its activation. Hydroxyapatite column chromatography revealed that the PKC in the major peak is type III. These results indicate that in lung cancer cells type III PKC activity is often elevated compared to other types of cancer cells. The growth of many lung cancer cell lines was inhibited by TPA.
Demethylation by 5‐Azacytidine Results in the Expression of Hepatitis B Virus Surface Antigen in Transgenic Mice
Authors: Araki Kimi , Miyazaki Jun‐ichi , Tsurimoto Toshiki , Inomoto Takeaki , Iwanaga Tomohisa , Matsubara Kenichi , Yamamura Ken‐ichi
Published: August 2005

In 14p3HB transgenic mice, which carry three tandem copies of hepatitis B virus (HBV) DNA, the HBV DNA was significantly methylated and no viral proteins were produced. To analyze the causal relationship between hypermethylation and gene inactivity, 5‐azacytidine was injected into the mice to demethylate HBV DNA. When postnatal 14p3HB mice were treated with the drug, hepatitis virus surface antigen was produced in these mice by 3 weeks of age, and the integrated HBV DNA of the liver was less heavily methylated. Our results suggest that injection of 5‐azacytidine can be used to efficiently activate a silent transgene such as HBV DNA in transgenic mice.
Advantages and Limitations of Stereological Estimation of Placental Glutathione S‐Transferase‐positive Rat Liver Cell Foci by Computerized Three‐dimensional Reconstruction
Authors: Imaida Katsumi , Tatematsu Masae , Kato Toshio , Tsuda Hiroyuki , Ito Nobuyuki
Published: August 2005

The applicability to a medium‐term bioassay for liver carcinogens of mathematical formulae for the calculation of numbers of foci per volume was examined in F344 rats. Two weeks after initiation with diethylnitrosamine, animals were given test compounds for 6 weeks, partial hepatectomy being performed at week 3. At week 8, the rats were killed, the livers removed and stained immuno‐histochemically for assessment of glutathione S‐transferase P form (GST‐P)‐positive foci development. Numbers and areas of lesions were measured two‐dimensionally using a color image analyzer, and the Enzmann and Campbell formulae for estimation of number and volume per cm3 were applied to the results. In addition, three‐dimensional reconstruction of individual foci was performed using up to 150 GST‐P stained foci, with the aid of a computerized graphic system. Both two‐ and three‐dimensionally expressed quantitative results were found to adequately demonstrate the modifying potential of test chemicals on hepatocarcinogenesis. The three‐dimensional approach was only more accurate if most of the foci were small and the liver was enlarged by compound treatment. Stereological reconstruction revealed that the shape of GST‐P‐positive foci, especially if relatively large, is not always spherical but that many demonstrate irregular branching forms, so that the assumptions behind Stereological estimation are not met. The results therefore show that care must be taken in applying mathematical formulae for the calculation of three‐dimensional data.
Gene Expression of Terminal Deoxynucleotidyl Transferase in Neoplastic Cells of Leukemia and Lymphoma
Authors: Oiwa Kazuko , Koiwai Osamu , Kaneda Tsuguhiro
Published: August 2005

Expression levels of terminal deoxynucleotidyl transferase (TdT) mRNA in fresh leukemia and lymphoma cells were measured by northern blotting analysis. Bands of 2.1 kb mRNA were detected in all of eight cases of TdT activity‐positive leukemias: two cases of null‐cell acute lymphoblastic leukemia (null‐ALL), two of common ALL, one of pre‐B ALL, one of T‐ALL, and two of chronic myelogenous leukemia in blastic crisis. One of the null‐ALL and one of the common ALL cases also showed large TdT mRNA (3.3 kb). Since all TdT activity‐positive samples exhibited TdT mRNA, the TdT gene might be mainly regulated at the transcription level in leukemic cells. An elevated level of 2.1 kb TdT mRNA was also detected in one lymphoma case, where neither TdT activity nor immunoreactive TdT was detected. The extensive chromosomal abnormality demonstrated in this case might be associated with the translational anomaly of TdT.
Detection of Granulocyte Colony‐stimulating Factor Produced by a Newly Established Human Hepatoma Cell Line Using a Simple Bioassay System
Authors: Tohyama Kaoru , Yoshida Yataro , Kubo Akemi , Sudo Tetsuo , Moriyama Masami , Sato Hisao , Uchino Haruto
Published: August 2005

A colony‐stimulating factor(CSF)‐producing tumor cell line (KX‐87) was established from a patient with hepatocellular carcinoma and marked granuloeytosis. This cell line formed tumors on nude mice in high frequency and the mice revealed marked granuloeytosis. In clonogenic assays of human bone marrow cells, KX‐87 conditioned medium (CM) supported the formation of colonies mainly consisting of neutrophilic granulocytes but had no burst‐promoting activity. The molecular weight of the colony‐stimulating activity (CSA) in KX‐87CM was estimated about 25,000 daltons by gel filtration and a new bioassay system. In principle, a subline of murine hemopoietic cell line NFS‐60 was cloned which was dependent on KX‐87CM. Then the growth of this subline was examined by a rapid and sensitive colorimetric tetrazolium assay. From these results, it was concluded that the CSA which KX‐87 cell line produced was G‐CSF.
Serial Section Study of Colonic Adenomas with Special Reference to Minute Carcinoma
Authors: Muto Tetsuichiro
Published: August 2005

A total of 73 colonic adenomas, 20 with mild atypia, 26 with moderate atypia and 27 with focal carcinoma (severe atypia), were examined by complete serial section in order to find the distribution of various degrees of atypia in adenomas. In serial sections minute carcinomatous foci were found in 3 (15%) and 12 (46.2%) of the adenomas with mild and moderate atypia, respectively. Fourteen adenomas (51.9%) with focal carcinoma had multiple foci of minute carcinoma elsewhere. Adenomas less than 1 cm in diameter showed 27% malignancy rate in serial sections. From this observation it is postulated that the malignant potential of colonic adenomas is much higher than previously believed.
Preventive and Antiproliferative Effects of Tumor Necrosis Factor against Experimental Hepatic Metastases of Mouse Colon‐26 Tumor
Authors: Nishiyama Yoshitaka , Fuchimoto Sadanori , Orita Kunzo
Published: August 2005

This investigation was undertaken in order to assess both the preventive and antiproliferative effects of tumor necrosis factor (TNF) in a hepatic metastasis model, by means of inoculation of mouse colon‐ 26 tumor cells into the portal vein via the superior mesenteric vein in male CDF1 mice, aged 5 weeks. Continuous 10‐day administration of natural human TNF‐α (nHuTNF‐α) following the tumor cell inoculation caused no reduction but rather an increase in the number of hepatic metastases. However, pretreatment with this preparation daily for 10 days before the inoculation caused a remarkable decrease in the number of hepatic metastases. This prophylactic effect was reversed by the intravenous administration of anti‐asialo GM1 antibody 24 h before the inoculation. The result of immunoperoxidase staining of liver specimens suggested that organ‐associated natural killer cells might play a role in the metastatic inhibition. An apparent antiproliferative effect on metastatic liver tumors was also recognized following injection of nHuTNF‐α from the 10th day after the inoculation. Thus, TNF appears to have important effects upon the host immune system, acting against liver metastases.
In vivo Effects of Recombinant Interferon Alpha A/D Incorporated in Gelatin Microspheres on Murine Tumor Cell Growth
Authors: Tabata Yasuhiko , Uno Kazuko , Muramatsu Shigeru , Ikada Yoshito
Published: August 2005

Intraperitoneal (ip) injections of gelatin microspheres containing a very small amount of recombinant human interferon alpha A/D (A/D‐IFN) (IFN‐microspheres) plus free A/D‐IFN improved the survival of mice bearing ascitic Meth A‐R1 cells which we had isolated as IFN‐resistant cells under in vitro conditions. The dose of free A/D‐IFN in one injection was 10,000 IU, which was insufficient by itself for manifesting in vivo antitumor activity. In these mice, in vivo Rl cell growth was suppressed and macrophage recruitment was enhanced in comparison with mice receiving other control agents. Administration of IFN‐microspheres alone was also effective but less than that of IFN‐microspheres plus free A/D‐IFN. Peritoneal macrophages obtained from normal or R1‐bearing mice receiving ip injection of IFN‐microspheres with or without free A/D‐IFN were activated to inhibit the in vitro growth of R1 cells. The intratumoral injection of IFN‐microspheres strongly inhibited the growth of solid R1 tumors. Intravenous injection of IFN‐microspheres was effective in preventing the pulmonary metastasis of B16 melanoma cells. These results indicate that the IFN‐microsphere is much more effective against tumors than free A/D‐IFN.
Construction of Mouse A9 Clones Containing a Single Human Chromosome Tagged with Neomycin‐resistance Gene via Microcell Fusion
Authors: Koi Minoru , Shimizu Motoyuki , Morita Hiroyuki , Yamada Hideto , Oshimura Mitsuo
Published: August 2005

Normal human fibroblasts (MRC‐5 or NTI‐4) were transfected with pSV2‐neo plasmid DNA. Fifty G418‐resistant fibroblast clones were isolated and independently fused to mouse A9 cells. The cell hybrids were selected and isolated in the medium containing G418 plus ouabaln. Since micronuclei were more efficiently induced in these hybrids compared to parental human fibroblasts by colcemid treatment, the transfer of neo‐tagged human chromosomes in the hybrids to mouse A9 cells was performed via microcell fusion. Two hundred A9 microcell hybrids were isolated and karyotyped. Among them, thirteen microcell clones, each containing a single human chromosome 1, 2, 5, 6, 7, 8, 10,11,12,15,18,19 or 20 were established. Isozyme analyses confirmed the presence of each human chromosome in these A9 microcell clones. The results of Southern blot and chromosomal in situ hybridization analyses indicate that the human chromosomes in these clones were tagged with pSV2‐ neo plasmid DNA.
Induction of Uterine Hemangioendothelioma and Lymphoma in (C57BL/6N×C3H/2N)F1 Mice by Oral Administration of Azathioprine
Authors: Ito Akihiro , Mori Masahiro , Naito Masashi
Published: August 2005

The tumorigenicity of 6‐(l‐methyl‐4‐nitro‐5‐imidazolyl)mercaptopurine (azathioprine), an imniuno‐suppressant, was examined in (C57BL/6N×C3H/2N)F1 mice. Animals were divided into 6 groups with 50 mice in each group, and they were given powdered diet mixed with 0, 5 or 20 ppm azathioprine starting at 6 weeks and ending at 100 weeks of age. Female mice, given 20 ppm azathioprine, developed lymphomas and uterine hemangioendotheliomas at incidences of 12.5 and 14.6%, respectively, which were significantly higher than the incidences in control mice (P< 0.05), Lymphoma and uterine hemangioendothelioma developed independently in different mice. On the other hand, the male mice given 20 ppm azathioprine had a significantly (P< 0.05) lower incidence of hepatic tumor (0.5%) compared to the control mice (16%).
Establishment and Characterization of Mouse Leukemia Cell Lines L615K and L7212K Derived from Transplantable Murine Leukemias Maintained in China
Authors: You Shengguo , Maeda Sakan , Murao Shin‐ichi , Takahashi Rei , Ishikawa Jiro , Miyazawa Masaaki , Nose Masato , Sugiyama Taketoshi
Published: August 2005

Two culture cell lines L615K and L7212K were established from transplanted murine leukemias L615 and L7212, which had been established and maintained in China for years. Based on morphological, immunological and gene rearrangement analyses, L7212K cells are considered to be of T‐cell origin while L615K cells might be immature T‐cells. Immunofluorescence assays of viable leukemia cells and fluorescence focus assays of their culture supernate for infectious viruses suggested that recombinant mink cell focus‐inducing viruses were significantly involved in both leukemic cell lines. Chromosome analysis of the L615K cells revealed a translocation t(12;17) which probably involved the c‐fos locus on chromosome 12, since the DNA rearrangement of c‐fos was demonstrated by Southern blot analysis with Hind III, and c‐fos has been assigned to this chromosome. Although the expression of this gene was not detected by RNA Northern blot analysis, c‐myc was slightly expressed in both L615K and L7212K cells.