bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of Cytosolic Phosphoethanolamine and Ethanolamine and Their Correlation with Prognostic Factors in Breast Cancer
Authors: Kano‐Sueoka Tamiko , Watanabe Toru , Miya Toshimichi , Kasai Hiroshi
Published: August 2005

Availability of accurate prognostic factors is vital in making decisions on cancer therapy. We have measured the cytosolic contents of phosphoethanolamine and ethanolamine in tumor tissues of 53 breast cancer patients in an attempt to explore the possibility that these amines could be used as prognostic indicators. The levels of phosphoethanolamine and ethanolamine were determined by high‐performance liquid chromatography. The ratios of the molar quantity of these amines or amino acids to that of alanine plus tyrosine, which eluted as a single peak, were used to analyze and compare the results among different tumor samples. The results indicated that the values for phosphoethanolamine or ethanolamine varied significantly more than the values for amino acids, such as glycine plus threonine or glutamine plus serine (these amino acids were eluted as single peaks, respectively). The values for phosphoethanolamine, ethanolamine, and phosphoethanolamine plus ethanolamine were analyzed in relation to several commonly used prognostic factors of breast disease. The results indicated that groups having higher mitotic indices had significantly higher values for phosphoethanolamine or phosphoethanolamine plus ethanolamine than the group having lower mitotic indices. As the stage of the disease increased, the values for phosphoethanolamine plus ethanolamine also seemed to become higher. No correlation, however, was observed between steroid hormone receptor positive and negative groups or between positive and negative groups with regard to involved axillary lymph nodes. The content of phosphoethanolamine or phosphoethanolamine plus ethanolamine in cytosol therefore seems to be correlated with some prognostic indicators.
Characterization of the Individual and Cross‐reactive Antigens Involved in the Anti‐tumor Immunity Induced by Use of an H‐2K‐erbB Recombinant Gene Transfectant
Authors: Ding Lin‐na , Yoshida Tomoaki , Isobe Ken‐Ichi , Rahman S. M. Jamshedur , Nagase Fumihiko , Yokochi Takashi , Kawashima Kohei , Nakashima Izumi
Published: August 2005

The specificities of the antisera raised in the CDF4 mice that had been immunized with the Pl. HTR tumor cells xenogenized by transfection with recombinant H‐2Kb‐erbB gene were studied. The antisera cross‐reacted with a broad range of tumor cell lines maintained either in vitro or in vivo in an immunofluorescence assay. However, they did not react at all with syngeneic normal tissue cells from thynms, spleen, bone marrow and fetal liver. Even though antigens related to the murine leukemia virus and murine mammary tumor virus (MuMTV) were demonstrated in many of the tumor cell lines tested with specific antibodies, these antigens did not seem to be primarily involved in the anti‐Pl. HTR antibody activity. The 74 kDa molecule, which was precipitated by the anti‐Pl. HTR anti‐serum from the surface radiolabeled cell extract of Pl. HTR tumor and was discriminated from the 70 kDa molecule precipitated by the anti‐MuMTV serum, was widely distributed among various tumor cell lines tested, but was absent in normal tissue cells. In contrast to the extensive cross‐reaction by the antibody, the cytotoxic T lymphocyte generated in the Pl. HTR immune mice were shown to be specific to the Pl. HTR tumor, and the 98 kDa molecule was precipitated by the anti‐Pl. HTR serum from the Pl. HTR tumor but not from other tumors tested. It is suggested from these results that the 98 kDa molecule is a candidate for an individual tumor‐specific transplantation antigen, and is immunodominant for inducing cytotoxic T lymphocytes to coexisting intrinsic retroviral antigens and other serologically cross‐reactive tumor antigens.
Malignant Cell Detection in Burkitt's Lymphoma Using Third‐complementarity‐determining Region (CDRIII), Clone‐specific Probe Developed by Sequencing DNA from Stored Slides
Authors: Asada Minoru , Miyashita Toshiyuki , Bessho Fumio , Kobayashi Noboru , Mizutani Shuki
Published: August 2005

The DNA sequence of the third‐complementarity‐determining region (CDRIII) of the immunoglobulin heavy chain (IgH) gene in a case of Burkitt's lymphoma was determined hy polymerase chain reaction (PCR) using template DNA extracted from a smear stored at room temperature for more than one year. The DNA sequence obtained from the stored slide was compared with that of DNA from a frozen lymph node biopsied at the initial presentation. The sequences were shown to he identical, implying that DNA from a smear on a stored slide can he used as a source of DNA for PCR amplification, sequencing, and development of a clone‐specific probe. Using oligonucleotides generated from one of the CDRIII sequences of the IgH gene as molecular probes, a retrospective study for the malignant clone on the smears was carried out. Malignant ceils were detectable in the peripheral blood at an early stage of hone marrow relapse but not in the peripheral blood or bone marrow at the initial presentation. No malignant clone was detected in the bone marrow when testicular infiltration was diagnosed by examination of a pathological specimen. Thus, the technique permits molecular analysis of hematologic malignancies of B‐cell lineage in cases where fresh or frozen specimens are not available.
A New CA125‐like Antigen (CA602) Recognized by Two Monoclonal Antibodies against a Newly Established Ovarian Clear Cell Carcinoma Cell Line (RMG‐II)
Authors: Nozawa Shiro , Yajima Masazumi , Sasaki Hirosuke , Tsukazaki Katsumi , Aoki Daisuke , Sakayori Motoko , Udagawa Yasuhiro , Kobayashi Toshifumi , Sato Ichiro , Furusako Shoji , Mochizuki Hiroshi
Published: August 2005

A cell line designated RMG‐II was established from the ascites of a patient with ovarian clear cell carcinoma. The chromosomal analysis revealed aneuploidy with a hypertetraploid modal numher and 8 marker chromosomes. Radioimmunoassay and immunocytochemical staining showed that RMG‐II cells produced some tumor markers such as CA125 and TPA. Two monoclonal antibodies, designated MA602‐1 and MA602‐6, were generated by immunization of mice with an extract prepared from the culture supernatant of RMG‐II cells. The epitopes recognized by these two monoclonal antibodies were proved to differ from the CA125 epitope, but to exist on the molecule bearing CA125. We developed a double‐determinant sandwich enzyme imnnmoassay using these two monoclonal anti‐bodies, and the antigen defined by this assay was termed CA602. CA602 was frequently found in the sera of ovarian cancer patients; the positive rates were 92%, 38%, 60%, and 80% for serous, mucinous, clear cell, and endometrioid ovarian carcinomas, respectively, when the cut‐off value was set at 60 U/ml (=mean + 3SD of healthy females). CA602 levels in serum were also high in endometriosis patients and in early pregnancy, as is the case for CA125, and the correlation coefficient between CA602 and CA125 was high (r=0.88). Our preliminary evidence suggests that this CA602 assay system has higher sensitivity than the CA125 one.
Detection of Bone‐type Alkaline Phosphatase by Monoclonal Antibodies Reacting with Human Osteosarcoma‐associated Antigen
Authors: Nagoya Satoshi , Uede Toshimitsu , Wada Takuro , Ishii Seiichi , Yamawaki Shinya , Kikuchi Kokichi
Published: August 2005

The antigen detected by monoclonal antibodies reacting with human osteosarcoma‐associated antigen was shown to he a phosphatidyl‐inositol (Pl)‐glycan‐anchored protein, which can be released from the cell surface by PI‐specific phospholipase C‐treatment. The antigen detected by 2D3 and 2H10 antibodies exhibited alkaline phosphatase activity. Both antibodies strongly reacted with bone‐type alkaline phosphatase. However, importantly, immunohistochemical analysis demonstrated that 2D3 and 2H10 did not react with alkaline phosphatase present in kidney or liver. In addition, neither placental nor intestinal alkaline phosphatase was recognized by 2D3 and 2H10 antibodies. These results indicated that two monoclonal antibodies, 2D3 and 2H10, are highly specific for bone‐type alkaline phosphatase and can distinguish bone alkaline phosphatase from liver alkaline phosphatase in spite of the fact that liver and bone alkaline phosphatase are encoded by the same gene.
Anti‐tumor Effects of Interleukin‐4 and Interleukin‐5 against Mouse B Cell Lymphoma and Possible Mechanisms of Their Action
Authors: Wu Hua‐Lang , Hirai Hisamaru , Inamori Ken , Kitamura Kiyoshi , Takaku Fumimaro
Published: August 2005

We investigated the anti‐tumor effects of recombinant mouse interleukin (IL)‐4 and IL‐5 by using a transplantable B cell lymphoma 38C13 cell line as a model. Daily local administration of either IL‐4 or IL‐5 produced moderate but significant inhibition of the rate of local tumor growth and prolongation of mean survival time (MST) in syngeneic C3H/HeJ mice; these anti‐tumor effects appeared to plateau at low doses. Histopathologic and immuno‐histochemical examination revealed necrotic changes in the cytokine‐treated tumors, associated with infiltration of inflammatory cells such as eosinophils, macrophages, and lymphocytes. The infiltrating lymphocytes were found to be Thy‐1.2+ T cells. To elucidate the importance of T cells, the rate of tumor growth and the MSTs were compared between athymic T cell‐deficient BALB/c nude mice and immunocompetent C3H/HeJ mice. In the nude mice the transplanted tumor grew more rapidly and the MST was shorter than in the normal mice, suggesting a significant contribution of infiltrating T cells in the anti‐tumor effects of the interleukins. Lastly, in vitro, growth inhibition of the 38C13 cells was observed in a dose‐dependent manner at relatively high concentrations of either cytokine. Therefore, we conclude that both IL‐4 and IL‐5 have moderate anti‐tumor effects against 38C13 B cell lymphoma both in vivo and in vitro, and that the observed in vivo anti‐tumor effects are probably mediated both by tumoristatic action of infiltrating cells, such as eosinophils, macrophages and T lymphocytes, and by direct anti‐proliferative action of the recombinant cytokines.
Experimental Metastasis of Oncogene‐transformed NIH 3T3 Cells in Chick Embryo
Authors: Endo Yoshio , Seiki Motoharu , Uchida Hiroyuki , Noguchi Mika , Kida Yuri , Sato Hiroshi , Mai Masayoshi , Sasaki Takuma
Published: August 2005

By means of a highly sensitive and quantitative assay for specific detection of metastasized tumor cells in chick embryonic organs using the polymerase chain reaction (PCR), we have examined the experimental metastatic ability of individual clones of NIH 3T3 cells, transformed with oncogenes: v‐Ki‐ras, v‐Ha‐ras, v‐src, v‐fos, and v‐abl. Such a transformed clone had different metastatic abilities in different embryonic organs. Among them, two clones of NIH 3T3 cells transformed with ras‐oncogenes (v‐Ki‐ras or v‐Ha‐ras) metastasized to liver and lungs of chick embryo, and grew there more rapidly than the other clones. The parental NIH 3T3 cells were detected as slight bands of PCR products after iv injection, indicating some cells were trapped in chick embryonic organs, but did not grow. These findings indicate that the transformed cells are able to invade the organ tissues and grow in embryonic chick organs, but non‐metastatic cells such as the untransformed‐NIH 3T3 cells are not able to grow in the secondary sites. These experiments clearly demonstrate the usefulness of this assay system to study genes involved in malignant transformation.
High Human IgG Levels in Severe Combined Immunodeficient Mouse Reconstituted with Human Splenic Tissues from Patients with Gastric Cancer
Authors: Kubota Tetsuro , Yamaguchi Hiroshi , Watanabe Masahiko , Yamamoto Takaaki , Takahara Tetsuya , Takeuchi Tooru , Furukawa Toshiharu , Kase Suguru , Kodaira Susumu , Ishibiki Kyuya , Kitajima Masaki
Published: August 2005

We implanted normal peripheral blood lymphocytes (PBL) from healthy donors and splenic tissues from patients with gastric cancers into the severe combined immunodeficient (SCID) mouse, demonstrating that SCID mouse with splenic tissue can produce a high level of human immuno‐globulin G (IgG). The normal PBLs at 107 and 108/mouse were implanted intraperitoneally, and three splenic tissues with a size of 3×3×3 mm from gastric cancer patients were inoculated subcutaneously into the bilateral backs of the mice. At 2, 4, 6 and 8 weeks after inoculation, mice were killed, and the human IgG was assessed by an ELISA method. SCID mice with splenic tissue revealed high human IgG levels from 2 weeks after inoculation and approximately 2 mg of IgG per ml was observed at 8 weeks post‐implantation, while the IgG levels in mice treated with PBLs were limited. Since the half life of the extrinsic human IgG was 10.2 days, the high level of human IgG in the SCID mice was supposed to be produced by human plasma cells in the splenic tissue from gastric cancer patients. This model was thought to be adequate for evaluating human immunological functions in vivo.
The Effect of Breast Self‐examination on Early Detection and Survival
Authors: Kuroishi Tetsuo , Tominaga Suketami , Ota Jun , Horino Toshio , Taguchi Tetsuo , Ishida Tsunehiro , Yokoe Takao , Izuo Masaru , Ogita Masami , Itoh Sueyoshi , Abe Rikiya , Yoshida Koichi , Morimoto Tadaoki , Enomoto Kohji , Tashiro Hideya , Kashiki Yoshitomo , Yamamoto Satoru , Kido Choichiro , Honda Kazuyoshi , Sasakawa Michizou , Fukuda Mamoru , Watanabe Hiromu
Published: August 2005

To investigate the effect of breast self‐examination (BSE), we compared the stages, survival, and the risk of death for 355 patients with breast cancer detected by BSE with those for 1,327 patients with breast cancer detected by chance. The early stages of the disease were found to be more common among the symptomatic breast cancer patients detected by BSE than those by chance. The 5‐year overall survival rate was 94.4% for the symptomatic patients detected by BSE, and was significantly higher by 8.7% than that (85.7%) for patients detected by chance (P< 0.001). The 10‐year survival rate was 81.6% for patients detected by BSE, and 76.6% for cases detected by chance (the difference was not significant). The overall difference between the two survival curves was statistically significant by the Iogrank test (P<0.01). A multivariate analysis using the Cox proportional hazards model showed that the risk of death for patients detected by BSE was smaller by 0.570 times than that for patients detected by chance, which was statistically significant (P< 0.05). The effect of biases inherent to BSE in the survival analysis cannot be controlled completely even after conducting multivariate analysis. These results suggest that BSE may contribute to the reduction of the risk of death through early detection of breast cancer. However, further examination should be conducted by other methods to obtain conclusive evidence.
In vivo Antitumor Effects of Fluoropyrimidines on Colon Adenocarcinoma 38 and Enhancement by Leucovorin
Authors: Iigo Masaaki , Nishikata Ken‐ichi , Hoshi Akio
Published: August 2005

Antitumor effect and active metabolites of fluoropyrimidines were examined in mice with transplant‐able colon adenocarcinoma 38 (Co 38). 5‐Fluoro‐2′‐deoxj uridine (FUdR) treatment resulted in a much higher level of free 5‐fluoro‐2′‐deoxyuridine‐5′‐monophosphate in the tumor than 5‐rluorouracil (5‐FU) did, and thymidylate synthase was almost completely inhibited after FUdR treatment, but FUdR showed weaker antitumor activity than 5‐FU did. Moreover, 5‐fluorouridine (FUR) also hardly inhibited tumor growth. A more marked tumor inhibition was obtained when FUdR and FUR were administered together. The antitumor activity of 5‐FU was similar to that of the combination of FUdR and FUR. In combination with 2,2’‐anhyilro‐5‐ethyluridine, a uridine phosphorylase inhibitor, FUdR lost its antitumor activity, but that of FUR was somewhat potentiated. On the other hand, in combination with leucovorin (LV), 5‐FU showed markedly potentiated antitumor activity, while the antitumor activity of FUdR or FUR was not potentiated. Addition of LV to the combination of FUdR and FUR enhanced the inhibitory effect of the drugs. From these results, the combination of FUdR and FUR together with LV, and the combination of 5‐FU and LV seem to be highly efficaceous against Co 38.
Radiosensitization and Hypoxic Cell Toxicity of NLA‐1 and NLA‐2, Two New Bioreductive Compounds
Authors: Papadopoulou Maria V. , Epperly Michael W. , Shields Donna S. , Bloomer William D.
Published: August 2005

Two new bioreductive compounds, 9‐[3‐(2‐nitro‐l‐imidazolyl)propylamino]acridine hydrochloride (NLA‐1) and 9‐[2‐(2‐nitro‐l‐imidazoryl)ethylamino]acridine hydrochloride (NLA‐2), have been prepared. They feature an acridine ring to intercalate with DNA, a 2‐nitroimidazole ring as the radiosensitizing moiety and an amino functionality for increased DNA‐binding and hydrophilicity. Time and concentration dependent cytotoxicity as well as radiosensitization efficacy of the two compounds under hypoxic or aerobic conditions were determined in vitro using V‐79 cells and an MTT colorimetric or clonogenic assay. The isosensitization point (ISP), defined as that drug concentration which results in the same survival decrement upon exposure of hypoxic or oxygenated cells to a given radiation dose, has been determined for both compounds at 7.5 Gy and the values are significantly lower than the ISPs of 5‐[3‐(2‐nitro‐l‐imidazolyl)propyl]phenanthridinium bromide, 2‐(2‐nitro‐l‐imidazolyl)ethylamine or misonidazole (MISO). NLA‐1 and NLA‐2 are potent hypoxic cytotoxins and on a concentration basis, more potent than MISO as radiosensitizers in vitro. The sensitization enhancement ratios were significantly increased when 1 h drug preincubation under hypoxia at 37°C was applied, before irradiation at room temperature.
Elevated Levels of Plasma Transforming Growth Factor‐β in Patients with Hepatocellular Carcinoma
Authors: Shirai Yasuhiro , Kawata Sumio , Ito Nobuyuki , Tamura Shinji , Takaishi Kenji , Kiso Shinichi , Tsushima Hirofumi , Matsuzawa Yuji
Published: August 2005

We measured the plasma transforming growth factor‐β (TGF‐β) concentration in 14 patients with human hepatocellular carcinoma (HCC) and 9 age‐matched normal subjects using growth inhibition assay of mink lung epithelial cells. The calculated plasma TGF‐β concentration in the patients with HCC was 28.6 ± 27.9 ng/ml (mean± SE), showing significant elevation compared with that in 9 normal subjects (5.3 ± 3.3 ng/ml, P<0.01). In three cases, we could measure plasma TGF‐β levels before and after their treatment for HCC. The plasma TGF‐β levels decreased from 59.0 to 18.2 ng/ml after hepatic resection in one case, and from 24.0 to 10.7 ng/ml and from 12.4 to 3.4 ng/ml after transhepatic arterial embolization in the other two cases. These data indicate that plasma TGF‐β level is elevated in patients with HCC, probably due to release from HCC tissues.
Transition of Phenotypic Dimorphism with Regard to Spontaneous Sister Chromatid Exchange in Epstein‐Barr Virus‐transformed Bloom's Syndrome Lymphoblastoid Cell Lines
Authors: Tatsumi Kouichi , Kurihara Takayuki , Arita Izumi , Tatsumi‐Miyajima Junko
Published: August 2005

We recently established four lymphoblastoid cell lines (LCLs) by infecting the peripheral blood of four Japanese patients suffering from Bloom's syndrome (BS) with Epstein‐Barr virus (EBV). During the course of propagating these cell lines, two of them exhibited dimorphism regarding spontaneous sister chromatid exchange (SCE), i.e., a mixed population consisted of cells with extremely high SCE levels characteristic of BS and cells with low SCE levels indistinguishable from that of normal control cells. On the other hand, the other two cell lines maintained a monomorphic population with high SCE levels at least until 30 weeks after EBV infection. The proportion of the cells with high SCE levels in the cell lines with dual phenotype declined as the population doubling numbers (PDN) increased with time and they became ultimately undetectable. The proportion of cells with low SCE levels at the time of EBV infection was estimated in one of these LCLs as 0.075% by extrapolating the linear regression of the logit for the proportion plotted against PDN. In view of the well‐known stability of the monomorphic phenotype in representative BS LCLs during extended cultivation, together with the present observations on the dual phenotype, we conclude that the frequent establishment of BS LCLs exclusively with low spontaneous SCE levels is attributable to the various proportions of low‐SCE cells existing in vivo in the B‐lymphocytes pool of BS individuals and to the selective pressure against the high‐SCE cells in in vitro cultures.
Granulocyte‐colony‐stimulating Factor Enhances the Circulating Hematopoietic Progenitors in Lung Cancer Patients Treated with Cisplatin‐containing Regimens
Authors: Mukai Jun‐nosuke , Shimizu Eiji , Ogura Takeshi
Published: August 2005

A phase II study examining the effects of human recombinant granulocyte‐colony‐stimulating factor (G‐CSF) on the growth of colony‐forming unit‐granulocyte macrophage (CFU‐GM) in the bone marrow and in the peripheral blood was performed in lung cancer patients treated with cisplatin‐containing regimens. Treatment with G‐CSF following chemotherapy significantly increased the absolute granulocyte count. No significant effect of G‐CSF on either the platelet or the red blood cell count was observed. Treatment with G‐CSF did not affect the CFU‐GM levels in the bone marrow, but did have a significant effect on peripheral blood CFU‐GM levels 14 days after initiation of chemotherapy (J°<0.05). Four patients demonstrated a rebound increase in the level of peripheral blood CFU‐GM during the first course of chemotherapy without G‐CSF. In contrast, eight patients displayed increase in peripheral blood CFU‐GM levels during the second course of chemotherapy with G‐CSF treatment. These findings demonstrate that G‐CSF is a potent stimulator of granulocyte proliferation as well as a potent agent for promoting transport of hematopoietic progenitors from the bone marrow into the peripheral blood.
Protection of Survival and Hematopoiesis in Irradiated Mice by 5‐FIuorouracil
Authors: Kojima Eiichi , Tsuboi Atsushi
Published: August 2005

The effects of whole‐body irradiation on survival and the hematopoietic system were studied in mice treated with 5‐fluorouracil (5‐FU). Animals (ddY‐SLC male mice, 8–10 weeks old) were injected with 5‐FU (i.p.) as a single dose (150 mg/kg) at various times before or after irradiation with X‐rays at graded doses (4.8 to 7.6 Gy). The treatment of mice with 5‐FU 5 days before irradiation was the most effective for the reduction of radiation lethality, having a radioprotective effect. The dose reduction factor (DRF) was 1.24. However, treatment with 5‐FU at 1 day and 2 hours before, or at various times after irradiation significantly increased the radiation lethality compared to the untreated controls, creating a radiosensitizing effect. The decrease or the increase of radiation lethality exhibited by 5‐FU was similar to. the radiation‐dose relationship pattern shown by endogenous and exogenous CFU‐S. The pattern of change of thrombocyte counts in the circulating blood after irradiation was greatly modified by pretreatment with 5‐FU 5 days before irradiation, effectively lessening the radiation‐induced depression. In contrast, the post‐irradiation patterns of leukocyte and erythrocyte variation did not show any significant change due to pretreatment with 5‐FU.
Detection of 2‐Amino‐1‐methyl‐6‐(4‐hydroxyphenyl)imidazo[4,5‐b]pyridine in Broiled Beef
Authors: Kurosaka Reiko , Wakabayashi Keiji , Ushiyama Hirofumi , Nukaya Haruo , Arakawa Nobuhiko , Sugimura Takashi , Nagao Minako
Published: August 2005

2‐Amino‐1‐methyl‐6‐(4‐hydroxyphenyl)imidazo[4,5‐b]pyridine (4′‐OH‐PhIP) showed mutagenicity in Salmonella typhimurium TA98 in the presence of S9 mix, inducing 180 revertants per 100 μg. Since creatinine, tyrosine and glucose, which are present in meat, are expected to be involved in the formation of 4′‐OH‐PhIP, its presence in broiled beef was examined. 4′‐OH‐PhIP was detected in broiled beef by high‐performance liquid chromatography and UV‐spectrometry after extraction with 0.1 N HCl and purification by blue cotton treatment and ion exchange column chromatography. Its level was estimated to be 21.0 ng per g of broiled beef, which is comparable to that of 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine (PhIP).
Enhancing Effects of Harman and Norharman on Induction of Preneoplastic and Neoplastic Kidney Lesions in Rats Initiated with N‐Ethyl‐N‐hydroxyethylnitrosamine
Authors: Hagiwara Akihiro , Sano Masashi , Asakawa Emiko , Tanaka Hikaru , Hasegawa Ryohei , Ito Nobuyuki
Published: August 2005

The modifying potential of two nephrotoxic agents, harman and norharman, on N‐ethyl‐N‐hydroxyethylnitrosamine (EHEN)‐induced renal and hepatic carcinogenesis was investigated in male F344/DuCrj rats. Animals were given 0.1% EHEN in their drinking water for the first 2 weeks as an initiator. Subsequently, starting 3 weeks from the commencement, they were fed diet containing these compounds at concentrations of 1000, S00 or 0 ppm until week 26, and then killed for light microscopic examination. The mean numbers of renal tubular cell hyperplasias/cm2 and those of tumors/cm2 in rats given harman and norharman at 1000 ppm after initiation, but not at 500 ppm, were significantly increased as compared to the control values. However, neither compound modified liver carcinogenesis. It is concluded that harman and norharman show enhancing effects on rat kidney carcinogenesis, when ingested at dose levels which cause renal tubular damage.
Clonal Origin of Skin and Bone Tumors Produced by Repeated Beta‐irradiation in Mosaic Cell Mice
Authors: Ootsuyama Akira , Tanaka Kazuhiko , Tanooka Hiroshi
Published: August 2005

Clonal origin of skin and bone tomors produced by repeated beta‐irradiation was determined by using mice with cellular mosaicism created by random X‐chromosome inactivation, on the basis of phosphoglycerate kinase‐1 (PGK). The backs of female C3H/He (Pgk‐1a/Pgk‐1b) mice were exposed to beta rays from 90Sr‐90Y at a dose of 3 Gy per exposure 3 times weekly until tumors appeared. The cumulative tumor incidence reached 100% 500 days after the beginning of irradiation, as determined by the Kaplan‐Meier method. All 8 tumors examined were of a single PGK phenotype: 5 squamous cell carcinomas and 2 osteosarcomas of A‐type, and 1 squamous cell carcinoma of B‐type. The absence of double PGK phenotype (AB‐type) tumors indicated the monoclonal origin of the tumors produced by repeated irradiation.
Maternal Transmisson of HTLV‐1 Other than through Breast Milk: Discrepancy between the Polymerase Chain Reaction Positivity of Cord Blood Samples for HTLV‐1 and the Subsequent Seropositivity of Individuals
Authors: Kawase Ken‐ichiro , Katamine Shigeru , Moriuchi Ryozo , Miyamoto Tsutomu , Kubota Kyoko , Igarashi Hisanaga , Doi Hiroshi , Tsuji Yoshiro , Yamabe Toru , Hino Shigeo
Published: August 2005

We used a nested polymerase chain reaction (PCR) to diagnose HTLV‐1 carriers. The DNA isolated from the nuclear extract obtained from frozen whole blood was found appropriate for PCR study both qualitatively and quantitatively. The use of freshly frozen whole blood made the field work much easier, and the use of a nuclear extraction procedure allowed DNA isolation in just 4 microcentrifuge tubes. We could not attain sufficient sensitivity to detect a single molecule with single‐step PCR, but nested PCR was confirmed to detect a single molecule/reaction. All samples of the seropositive group including 94 blood donors, 66 mothers, and 13 children were positive in the nested PCR, while none of the seronegative group, including 198 blood donors and 285 children, was positive. Although 18/717 (2.5%) cord blood samples obtained from babies born to carrier mothers were PCR‐positive, none of 5 formula‐fed children tested who had been PCR‐positive in the cord blood gave evidence of infection later on. Furthermore, all of 4 seropositive infected children who were formula‐fed had been PCR‐negative in their cord blood. The results are not consistent with intrauterine infection, but suggest the presence of a perinatal or postnatal infection route other than through breast milk.
Pulmonary Large Cell Carcinoma Expressing Neuroendocrine Markers: The Morphological, Biological, and Neuroendocrine Features of Their Cell Lines and Surgical Cases
Authors: Kasai Kiyoshi , Kameya Tom , Kawakubo Yasuaki , Sato Yuichi , Wada Chieki , Itoh Hiroshi
Published: August 2005

A cell line expressing neuroendocrine (NE) markers, designated as KTS9, was established from a human large cell carcinoma of the lung using serum‐free medium, ACL‐3. KTS9 cells showed morphological characteristics of large cell undifferentiated carcinoma (LGUC) and expressed some general NE markers including neuron‐specific enolase (NSE), protein gene product (PGP) 9.5, neural cell adhesion molecule (N‐CAM), synaptophysin and neurofilaments (NF) of 200 kd. Some cells of this cell line were positive to chromogranin‐A (CG‐A), but did not express Leu7 or aromatic L‐amino acid decarboxylase (AADC). Such a cell line derived from LCUC with NE properties has not previously been reported. The biological and NE properties of the KTS9 cell line were compared with those of 2 surgical cases of LCUC with NE markers and of the KTA7 cell line previously reported to derive from large cell carcinoma and to possess NE markers such as alpha‐hCG, PGP9.5 N‐CAM and AADC. Tumor cells of 2 large cell carcinomas expressed NSE, PGP9.5, N‐CAM and NF. The KTS9 and KTA7 cell lines and 2 large cell carcinomas were thus considered to be LCUCs with NE differentiation. Both lines had the morphological characteristics of LCUC, relatively short doubling time and discordant expression of NE markers, indicating them to be closely related to the variant type of small cell carcinoma cell lines and thus possibly to represent high‐grade malignancy. They may be useful for examining the biological behavior and NE features of large cell‐type NE tumors of the lung.
Significance of Strain and Sex Differences in the Development of 252Cf Neutron‐induced Liver Tumors in Mice
Authors: Ito Akihiro , Takahashi Tadateru , Watanabe Hiromitsu , Ogundigie Peter Osa , Okamoto Taro
Published: August 2005

Mouse liver tumors occurring in C3H/HeN, C57BL/6N and C3B6F1 hybrid (C3H × C57BL) were studied following 252Cf fission neutron irradiation. Three strains of mice of both sexes (about 30 mice/group) were irradiated once with 252Cf at doses of 0,12.5, 50 and 200 cGy. The groups were observed for 13 months after irradiation. The incidence of liver tumors in the non‐irradiated controls was 0% in both sexes of CS7BL/6N, 11.7% in males and 0% in females of C3B6F1 and 39.5% in males and 11.4% in females of C3H/HeN mice. In the four strains of mice thus far studied, including B6C3F1 hybrid (CS7BL × C3H) which was previously studied, 252Cf irradiation has increased the tumor incidence dose‐dependently in males and in females, but less effectively in females. The mean number and size of liver tumors were clearly correlated with tumor incidence. The incidence was always highest in C3H/HeN mice of both sexes, followed by B6C3F1, C3B6F1 and C57BL/6N mice. The influence of sex hormones was studied in B6C3F1 mice of both sexes after 200 cGy of 252Cf irradiation. In males, the incidence of liver tumors was significantly decreased from 55.2% to 23.3% and 25.9% after orchidectomy, and in females it was slightly decreased from 27.6% to 14.8% and 18.8% after ovariectomy. Supplementation of testosterone in orchidectomized mice did not restore the occurrence of liver tumors.
Altered Glycosylation of Membrane Glycoproteins Associated with Human Mammary Carcinoma
Authors: Hiraizumi Sen , Takasaki Seiichi , Ohuchi Noriaki , Harada Yuko , Nose Masato , Mori Shozo , Kobata Akira
Published: August 2005

N‐Linked sugar chains of normal mammary gland, mammary carcinomas (primary lesion), and axillary lymph node metastases of mammary carcinomas were released from their membrane preparations by hydrazinolysis and their structures were analyzed. Fractionation using a Datura stramonium agglutinin (DSA)‐Sepharose column revealed that the metastasized carcinomas contain more than twice as much DSA‐binding oligosaccharides as the normal gland, and the primary carcinomas contain an intermediate amount. These oligosaccharides were elucidated to have tri‐ and tetraantennary structures containing the GlcNAcβ1 → 6(GlcNAcβ1 → 2) Man group with and without N‐acetyllactosamine repeating units. Lectin blot analysis of membrane glycoproteins and histochemical staining of tissues using biotinylated DSA indicated that these glycosylation changes predominantly occur in a limited number of glycoproteins with apparent molecular weights of 90,160, and 210 kilodaltons, and mammary carcinomas are distinguishable from normal gland by their intense intracytoplasmic staining.
A Prospective Study of Atrophic Gastritis and Stomach Cancer Risk
Authors: Kato Ikuko , Tominaga Suketami , Ito Yoshiaki , Kobayashi Seibi , Yoshii Yuri , Matsuura Akira , Kameya Akira , Kano Tomoyuki , Ikari Akira
Published: August 2005

The relation of atrophic gastritis, other gastric lesions and lifestyle factors to stomach cancer risk was prospectively studied among 3,914 subjects who underwent gastroscopic examination and responded to a questionnaire survey at the Aichi Cancer Center Hospital. During 4.4 years of follow‐up on average, 45 incident cases of stomach cancer were identified at least three months after the initial examination. If the baseline endoscopic findings indicated the presence of atrophic gastritis, the risk of developing stomach cancer was increased 5.73‐fold, compared with no indication at the baseline. The risk further increased with advancing degree of atrophy and increasing extension of atrophy on the lesser curvature. These trends in the relative risks were statistically significant (P= 0.027 andP= 0.041, respectively). The risk of developing stomach cancer was statistically significantly increased among subjects with gastric polyps, but not among those with gastric ulcer. Stomach cancer cases tended to consume more cigarettes, alcohol, rice, pickles and salted fish gut/cod roe and less fruits and vegetables and to have more family histories of stomach cancer than noncases, although these differences were not statistically significant. The results of the present study provide additional evidence on the relation between atrophic gastritis and stomach cancer and suggest a need for intensive follow‐up of patients with atrophic gastritis and gastric polyps.
Clinicopathologic Characteristics and Prognosis of Breast Cancer Patients Associated with Pregnancy and Lactation: Analysis of Case‐Control Study in Japan
Authors: Ishida Tsunehiro , Yokoe Takao , Kasumi Fujio , Sakamoto Goi , Makita Masujiro , Tominaga Takeshi , Simozuma Kohjiro , Enomoto Kohji , Fujiwara Kiyoshi , Nanasawa Takeshi , Fukutomi Takashi , Hirota Teruyuki , Fukuda Mamoru , Miura Shigeto , Koyama Hiroki , Inaji Hideo , Sonoo Hiroshi
Published: August 2005

Clinicopathologic characteristics and prognosis of breast cancer patients associated with pregnancy and lactation were clarified by means of a case‐control study of matched non‐pregnant and non‐lactating patients with breast cancer. From 18 institutions in Japan, a total of 192 subjects with breast cancer diagnosed during pregnancy (72 cases) and lactation (120 cases) were collected between 1970 and 1988, accounting for 0.76% of all breast cancer patients. The duration of symptoms was longer and tumor size was larger in the study subjects. Although the disease‐free interval was longer than that in the control patients, the survival time was shorter. There was no characteristic difference in histologic type. Vascular invasion and lymph node metastasis were found more frequently in the subjects. The positive rates of estrogen receptor and progesterone receptor were lower in the subjects. The 5‐ and 10‐year survival rates of the study patients were 65% and 55%, respectively, and these survivals were significantly lower than those of the control (P < 0.001). The survival rates were poorer in the subjects, in accordance with stage and lymph node metastasis. The results suggest that most of the patients with breast cancer diagnosed during pregnancy and lactation are in a more advanced stage because of a delay in detection and diagnosis, and hence have unfavorable prognosis. Therefore, it is important to diagnose and treat early for improvement of prognosis in patients with breast cancer during pregnancy and lactation.
Comparison of the Anatomic Distribution of Stomach Cancer and Precancerous Gastric Lesions
Authors: You Wei‐cheng , Blot William J. , Chang Yun‐sheng , Li Ji‐you , Jin Mao‐lin , Zhao Yongxing , Kneller Robert W. , Xie Yu‐quan , Zhang Lian , Xu Guang‐wei , Fraumeni Joseph F.
Published: August 2005

The anatomic distribution of precancerous gastric lesions among 3,400 residents in Linqu, Shandong Province of China, was compared with the anatomic distribution of stomach cancer (SC) among 959 patients in Tokyo, Japan. The incidence of SC is high in both areas, and locations within the stomach of the precancerous and malignant lesions were classified using similar criteria. Chronic atrophic gastritis (CAG) affected 98% of the population in Linqu, with intestinal metaplasia (IM) the most severe diagnosis in 33% and dysplasia (DYS) in 20%. Neither the SC nor precancerous lesions were uniformly distributed in the stomach. Among the DYS 3% were along the greater curvature of the body, 15% along the lesser curvature of the body, 25% in the angulus, 22% along the lesser curvature of the antrum, and 34% elsewhere in the antrum. Among the SC the corresponding percentages were 2, 16, 28, 25 and 29. The similarity to the SC distribution increased gradually from CAG to IM to DYS, providing further evidence for the multistage progression of precancerous gastric lesions.
Effects of Modifying Agents on Conformity of Enzyme Phenotype and Proliferative Potential in Focal Preneoplastic and Neoplastic Liver Cell Lesions in Rats
Authors: Tsuda Hiroyuki , Ozaki Keisuke , Uwagawa Satoshi , Yamaguchi Shuji , Hakoi Kazuo , Aoki Toyohiko , Kato Toshio , Sato Kiyomi , Ito Nobuyuki
Published: August 2005

Development of preneoplastic lesions in the rat liver under the influence of various modifiers was investigated with particular attention to changes in simultaneous expression of altered enzyme phenotype within the lesions (conformity) and proliferation potential. Degree of conformity of marker enzymes such as glutathione S‐transferase placental form (GST‐P), glucose‐6‐phosphate dehydrogenase (G6PD), glucose‐6‐phosphatase, adenosine triphosphatase and γ‐glutamyltranspeptidase was compared with levels of S‐bromo‐2‐deoxyuridine labeling. After initiation with diethylnitrosamine, rats were administered the hepatopromoter sodium phenobarbital (PB, 0.05%), the antioxidant ethoxyquin (EQ, 0.5%), or a peroxisome proliferator, clofibrate (CF, 1.0%) or di(2‐ethylhexyl)‐phthalate (0.3%) and killed at week 16 or 32. The PB promoting regimen was clearly associated with increase in the numbers of high conformity class lesions simultaneously expressing three to five enzymes, and elevated proliferation potential. The inhibitor, EQ, in contrast, brought about a time‐dependent decrease in conformity so that only 1 or 2 alterations were most commonly observed at week 32. Lesion populations in the peroxisome proliferator‐ and especially CF‐treated cases were characterized by obvious dissociation between degree of conformity and proliferative status. Such treatment‐dependent differences were not always correlated with the size of the lesion. The results thus suggested that the conformity and proliferation potential of preneoplastic lesions are dependent on modification treatment. Overall, GST‐P was found to be the most reliable marker, although G6PD was less influenced in the peroxisome proliferator cases.
Lack of Allelic Preference in Amplification and Loss of the c‐myc Oncogene in Methylcholanthrene‐induced Mouse Sarcomas
Authors: Niwa Ohtsura , Kominami Ryo
Published: August 2005

Sarcomas were induced in Fl mice between C57BL/6N and C3H/He strains by subcutaneous injection of methylcholanthrene. The c‐myc oncogene was found to be amplified in 16 cases among 43 sarcomas of C57BL/6N × C3H/He mice and 1 case among 5 sarcomas of the reciprocal cross. The origin of the amplified allele was determined by the polymerase chain reaction single strand conformation polymorphism analysis. Among the 17 sarcomas, only one had both of the alleles amplified. The rest of the tumors carried the amplified c‐myc allele coming either from C57BL/6N (9 cases) or from C3H/He (8 cases). These results indicate that the c‐myc allele is amplified randomly in methylcholanthrene‐induced mouse sarcomas irrespective of its origin, such as paternal or maternal allele and C57BL/6N or C3H/He allele. In addition to these changes, the unamplified c‐myc oncogene was found to be lost in 12 cases out of the 17 sarcomas with the amplification.
Vertical Transmission of HTLV‐1 in HTLV‐1 Carrier Rat
Authors: Ami Yoshihiro , Kushida Shigeki , Matsumura Masayuki , Yoshida Yuki , Kameyama Tsuneo , Sugiyama Yoshihiro , Yagami Ken‐ichi , Uchida Masako , Uchida Kazuhiko , Koiso Kenkichi , Miwa Masanao
Published: August 2005

A female F344 rat was injected with 2.4 × 106 MT‐2 cells intravenously at 3 and 4 weeks old, and was mated with a non‐infected male rat at the 17th week after injection, when the dam rat contained HTLV‐1 provirus in the peripheral blood mononuclear cells as determined by polymerase chain reaction. HTLV‐1 provirus was detected in at least 2 out of 9 offspring. This system should be useful for studies on the routes and prevention of vertical transmission and on elimination of once‐transmitted HTLV‐1.
Physical Ordering of Three Polymorphic DNA Markers Spanning the Regions Containing a Tumor Suppressor Gene of Renal Cell Carcinoma by Three‐color Fluorescent in situ Hybridization
Authors: Inazawa Johji , Ariyama Takeshi , Abe Tatsuo
Published: August 2005

Fluorescent in situ hybridization (FISH) is a powerful technique for gene mapping and multi‐color FISH allows us to determine directly the order of two or more probes on both metaphase and interphase chromosomes. We report the physical ordering of three DNA markers by three‐color FISH using two fluorochrome dyes, fluorescein isothiocyanate (FITC; green) and rhodamine (red). The third color was visualized as a pseudo‐color (yellow) generated by optical interference with FITC and rhodamine. Using this system we could rapidly determine the order of three polymorphic DNA markers located on the 3p23–p21.2 bands which span a tumor suppressor gene for renal cell carcinoma, lung carcinoma, and several other types of tumors.
Forestomach Neoplasm Induction in F344/DuCrj Rats and B6C3F1 Mice Exposed to Sesamol
Authors: Tamano Seiko , Hirose Masao , Tanaka Hikaru , Asakawa Emiko , Ogawa Kumiko , Ito Nobuyuki
Published: August 2005

Sesamol was administered at a dietary level of 2% to groups of 30 male and female F344/DuCrj rats and B6C3F, mice for 104and 96 weeks, respectively. Squamous cell carcinomas in the forestomach were Induced in nine of 29 (31%) effective male rats, three of 30 (10%) female rats, eleven of 29 (38%) male mice and five of 30 (17%) female mice treated with sesamol. Papillomas developed in ten of 29 (34%) male rats and fourteen of 30 (47%) female rats, but not in any of the mice. Hyperplasias developed in almost all rats and mice of both sexes. Significant differences from control values were found for all three lesions in rats and for carcinoma and hyperplasia categories in mice. The incidences of other tumors in the 2% sesamol group were comparable with control values. In conclusion, sesamol induces squamous cell carcinomas in the forestomach of rats and mice, males being more susceptible than females.
Absence of Activating Mutations in the Transmembrane Domain of the c‐erbB‐2 Proto‐oncogene in Human Lung Cancer
Authors: Sachse Richard , Murakami Yoshinori , Shiraishi Masahiko , Hayashi Kenshi , Sekiya Takao
Published: August 2005

The rat neu gene is known to be activated by a point mutation in its predicted transmembrane domain. Overexpression of the human homologue of neu, the c‐erbB‐2 gene, in human lung cancer has been reported, and a similar activating point mutation has been suggested. Therefore, we tested for possible aberrations of the c‐erbB‐2 gene in the region of the transmembrane domain in surgical specimens of human primary lung cancer from 190 patients, and also examined 24 metastases and 26 specimens of noncancerous portions of the lung of the same patients. Single‐strand conformation polymorphism analysis of polymerase chain reaction products revealed no point mutations in the target domain in any of these specimens.
Recombinant Human Interferon‐α2a Increases Hormone Receptor Level of a Human Breast Carcinoma Xenograft in Nude Mice and Enhances the Anti‐proliferative Activity of Tamoxifen
Authors: Josui Kazuya , Kubota Tetsuro , Kitajima Masaki
Published: August 2005

The effect of recombinant human interferon‐α2a (rhIFN‐α2a) on the hormone receptor level and antitumor activity of tamoxifen (TAM) was investigated in nude mice using ZR‐75‐1, an estrogen receptor (ER)‐positive, and progesterone receptor (PgR)‐negative human breast carcinoma xenograft. ER levels (maximum binding sites) of tumors treated with rhIFN‐α2a at a dose of 6 × 104 U/mouse/ day for 1 or 3 wk were not significantly different from the control, whereas those with rhIFN‐α2a at a dose of 6 × 104 U/mouse/day for 1 or 3 wk were higher than the control (3.9‐ to 4.4‐fold) with a significant difference at P < 0.01. The increase of ER by rhIFN‐α2a was investigated using a sucrose density gradient method. The peak was only seen at 8S in both rhIFN‐α2a‐treated tumor and control ER, and the sedimentation patterns were almost the same, suggesting that both ERs were essentially equivalent. On the other hand, PgR of all the treated groups could be detected, while that of the control group was undetectable. The antitumor effect of the combination treatment of rhIFN‐α2a and TAM was compared with those of single treatments. While rhIFN‐α2a at a dose of 6 × 105 U/mouse/day and TAM did not show a combination effect, rhIFN‐α2a at a dose of 6 × 104 U/mouse/day and TAM showed a synergistic combination effect, and ER was decreased to the threshold of detection by the combination treatment. These findings indicated that a low dose of rhIFN‐α2a increased the ER levels of ER‐positive human breast cancer in vivo as well as in vitro and enhanced the anti‐proliferative effect of TAM, and the newly synthesized ER was essentially the same as the original ER.
Pharmacokinetic Study of Mitomycin C with Emphasis on the Influence of Aging
Authors: Miya Toshimichi , Sasaki Yasutsuna , Karato Atsuya , Saijo Nagahiro
Published: August 2005

Mitomycin C (MMC) at a dose of 8 mg/m2 was administered by short intravenous infusion to 14 cancer patients. All patients had normal renal, hepatic, cardiac and bone marrow functions. The plasma level of MMC, which was determined by high‐performance liquid chromatography over a 24–h period after the infusion, fitted the 2‐compartment model curve except for one patient. There was a significant correlation between the area under the time versus concentration curve (AUC) of MMC and the age of patients (r= 0.558, P < 0.05). Pharmacokinetic parameters in one of the older (a 69‐year‐old male) patients were extremely different from those of the other 13 patients. This patient experienced severe myelosuppression, probably due to the markedly increased AUC of MMC. Our results suggest that a patient's age has a significant influence on the pharmacokinetics of MMC in patients with normal major organ functions and that in some patients, MMC pharmacokinetics may be altered, possibly due to interpatient variation in the activation or metabolic pathway of MMC.