bluuebunny/crossref_metadata_2025 huggingface dataset:
Synergistic Enhancement of Glutathione S–Transferase Placental Form–positive Hepatic Foci Development in Diethylnitrosamine–treated Rats by Combined Administration of Five Heterocyclic Amines at Low Doses
Authors: Hasegawa Ryohei , Shirai Tomoyuki , Hakoi Kazuo , Takaba Katsumi , Iwasaki Shogo , Hoshiya Toru , Ito Nobuyuki , Nagao Minako , Takashi Takashi
Published: August 2005

Potential synergism among 5 heterocyclic amines at low doses in the induction of glutathione S–transferase placental form (GST–P)–positive liver cell foci was examined in an 8–week experiment using male rats initially given diethylnitrosamine (200 mg/kg, ip). The heterocyclic amines applied were 3–amino–l–methyl‐5H–pyrido[4,3–b]indole (500 ppm), 2–amino–6–methyldipyrido[l,2–a:3′,2′–d]–imidazole (500 ppm), 2–amino–3–methyl–9H–pyrido[2,3‐b]indole (800 ppm), 2–amino‐9H–pyrido[2,3–b]indole (800 ppm), and 2–amino–l–methyl–6–plienyliniidazo[4,5–b]pyridine (PhIP, 400 ppm). Separate groups received each chemical at the dose used in earlier carcinogenicity assays (above doses), at 1/5 or 1/25 of these, or all 5 chemicals together, each at the 1/5 or 1/25 levels. The numbers and areas of GST–P–positive foci were significantly increased with all chemicals, except for PhIP, at the highest dose, the results being consistent with the reported liver carcinogenicity. In the combined treatment at the 1/5 dose levels, synergistic enhancement occurred; the numbers and areas of foci were significantly increased above the sums of individual data. However, this was not the case for the 1/25 dose groups. Although the synergism between pyrolysis products in liver carcinogenesis depended on the dose and combination of chemicals, the findings, together with those from a previous experiment using 5 different heterocyclic amines, are of particular significance since several heterocyclic amines might be simultaneously generated during cooking of foodstuffs.
Enhancing Potential of 6 Different Carcinogens on Multi‐organ Tumorigenesis after Initial Treatment with N‐Methyl‐N‐nitrosourea in Rats
Authors: Uwagawa Satoshi , Tsuda Hiroyuki , Inoue Tadashi , Tagawa Yoshiaki , Aoki Toyohiko , Kagawa Masataka , Ogiso Tadashi , Ito Nobuyuki
Published: August 2005

The advantages of applying a whole‐body concept to the assessment of carcinogenic potential of compounds in a two‐stage model after initiation by N‐methyl‐N‐nitrosourea (MNU) were investigated. Male, 6‐week‐old F344 rats were injected with MNU (20 mg/kg, i. p.) twice a week for 4 weeks and they then received 3,2′‐dimethyl‐4‐aminobiphenyl (DMAB) (50 mg/kg, s.c., once a week), N,N′‐dibutylnitrosaraine (DBN) (0.05%, in drinking water), N‐bis(2‐hydroxypropyl)nitrosamine (DHPN) (0.1%, in drinking water), diethylstilbestrol (DES) (2.5 ppm, in diet), sodium o‐phenylphenate (S. OPP) (2%, in diet) or captafol (0.15%, in diet) for 20 weeks. All six carcinogens enhanced the incidences of preneoplastic and neoplastic lesions in their respective target organs: liver, pancreas, small intestine and urinary bladder with DMAB; liver, esophagus, forestomach and urinary bladder with DBN; thyroid, lung, liver, esophagus, forestomach, small intestine and urinary bladder with DHPN; liver and forestomach with DES; and thyroid, forestomach, kidney and urinary bladder with S. OPP; liver and forestomach with captafol. The results suggested that prior treatment with MNU sensitized the tissues to the organotropic carcinogenic potential of chemicals given thereafter for as short a period as 20 weeks. Thus, this system could be utilized as a whole‐body medium‐term bioassay system for the screening of environmental carcinogens, bridging the gap between in vitro mutagenicity and long‐term carcinogenicity tests.
Genetic Alteration of p53 in Some Patients with Adult T‐Cell Leukemia
Authors: Nagai Hirokazu , Kinoshita Tomohiro , Imamura Jun , Murakami Yoshinori , Hayashi Kenshi , Mukai Kiyoshi , Ikeda Shuichi , Tobinai Kensei , Saito Hidehiko , Shimoyama Masanori , Kunitada Kunitada
Published: August 2005

Abnormalities of p53 mRNA in adult T‐cell leukemia (ATL) were analyzed using reverse transcription‐polymerase chain reaction‐single strand conformation polymorphism analysis. Mutations were present in two of 12 ATL patients studied, but not in 3 cell lines immortalized by human T cell leukemia virus type 1 (HTLV‐1) infection in vitro. Direct sequencing analysis of the p53 gene from these two patients revealed missense point mutations at codon 153 (arginine to histidine) or codon 220 (cysteine to tyrosine), respectively. Immunohistochemical analysis revealed the elevated expression of p53 proteins in ATL cells from a patient carrying the mutated p53 gene at codon 158. Neither gross rearrangement of p53 gene nor abnormal size of mRNA for the gene was demonstrated by Southern or Northern blot analyses. Thus, there is a mutated p53 in some patients with ATL. The involvement of abnormalities in some suppressor oncogenes may play a role in the development of ATL.
Effects of the Tumor Inhibitor IKP‐104, a 4(lH)‐Pyridinone Derivative, on Cytoskeletal Microtuhules of Cultured Tumor Cells
Authors: Mizuhashi Fukutaro , Murata Kyoji , Kitagaki Tadaharu , Isao Isao
Published: August 2005

The effects of IKP‐104, a 4(lH)‐pyridinone derivative, on the mitotic profile and cytoskeletal microtubule dynamics of cultured B16 melanoma cells were examined in order to investigate the mechanism of its antitumor activity. The exposure to IKP‐104 caused accumulation of cells in abnormal metaphase with chromosomes scattered within the cytoplasm and induced polyploid and multinucleate cells as detected by differential staining microscopy with brilliant blue R and safranin O. An immunofluorcsceiicc study with monoclonal anti‐α‐tubulin antibody revealed that IKP‐104 diminished cytoskeletal microtubules of both interphase and mitotic cells, resulting in induction of a few fragments resembling “microtubular bundles” induced by vinblastine (VLB). These results indicated that IKP‐104 arrests cells in the mitotic phase by inhibition of polymerization and induction of depolymerization of cytoskeletal microtubules, similarly to VLB.
Topoisomerase Inhibitors Have Potent Differentiation‐inducing Activity for Human and Mouse Myeloid Leukemia Cells
Authors: Nakaya Kazuyasu , Chou Sawako , Kaneko Motohisa , Nakamura Yasuharu
Published: August 2005

DNA topoisomerase inhibitors, camptothecin and 4′‐demethylepipodophyllotoxin ethylidene‐jS‐D‐glucoside (VP16) had strong differentiation‐inducing activity for all five kinds of leukemia cells examined (human HL60, U937, ML1, and K562 cells and mouse Ml cells) as judged from measurements of various differentiation markers. The characteristics that appeared as a result of differentiation induced by these inhibitors were essentially similar in every cell line. Exposure to VP16 for 2 h induced both differentiation and DNA‐strand breaks in K562 cells, whereas podophyl‐lotoxin, which lacks topoisomerase II inhibitory activity, induced neither differentiation nor DNA‐strand breaks in these cells. These results suggest a parallelism between the induction of differentiation and that of DNA‐strand breaks. The combination of VP16 and recomhinant tumor necrosis factor α (rTNFα) synergistically induced differentiation of human U937, ML1, and M1 cells and had an additive effect on HL60 cells. Simultaneous treatment with rTNFa plus camptothecin or VP16, or pretreatment with camptothecin or VP16, followed by rTNFα induced marked differentiation of Ml cells. These results indicate that inhibition of topoisomerase (either topoisomerase I or II) followed by the action of rTNFα was effective in inducing differentiation of leukemia cells.
Objective Quantification of Grade of Atypia in Epithelial Tumors of the Stomach by Image Processing
Authors: Ishido Tatsuya , Nakamura Kyoichi
Published: August 2005

An image processing technique to extract tubular and nuclear areas from histological specimens stained with hematoxylin was established. Two indices representing tubular density and nucleus‐to‐cytoplasm ratio of glands were examined in intestinal‐type tumor of the stomach. In specimens clearly stained with hematoxylin, significant differences of brightness among nuclei, cytoplasm and stroma can be obtained. Image processing by computer enabled us to identify nuclei, cytoplasm and stroma by utilizing the differences of brightness among them. Both indices tended to increase with severity of histological atypia and their values in the tubular adenoma group were significantly different from those in the tubular adenocarcinoma group (P< 0.05). Therefore, these two indices could be regarded as valid and objective measures of the grade of histological atypia. Eighty‐four percent of tubular adenoma and tubular adenocarcinoma cases were classified correctly with the discriminant formula and critical value calculated from the two indices on the basis of measurement of the images magnified by 40 times. There is, however, still a non‐negligible overlap of discriminant scores between the tubular adenoma group and tubular adenocarcinoma group. More indices representing histological atypia will be required to allow objective differential diagnosis between malignant and benign lesions.
Tumor Localization and in vivo Antitumor Activity of the Immunoconjugate Composed of Anti‐human Colon Cancer Monoclonal Antibody and Mitomycin C‐Dextran Conjugate
Authors: Noguchi Akinori , Takahashi Toshio , Yamaguchi Toshiharu , Kitamura Kazuya , Takakura Yoshinobu , Hashida Mitsuru , Sezaki Hitoshi
Published: August 2005

The tissue distribution and in vivo antitumor activity of a novel monoclonal antibody‐mitomycin C conjugate (A7‐MMCD) composed of anti‐human MAb A7 and MMC‐dextran conjugate were investigated using tumor‐bearing mice. A7‐MMCD was prepared via an anionic dextran intermediate for the purpose of keeping the non‐specific uptake by the reticuloendothelial system to a minimum, 111n‐labeled A7‐MMCD showed about a 5‐times‐greater accumulation in SW1116 (targeted tumor) than in S180 (non‐targeted tumor) 48 h after injection, and produced a tumor‐to‐blood ratio which was 3 times higher in SW1116‐bearing mice than in S180‐bearing mice 96 h after injection. Accumulations in the liver, spleen, and kidney were also observed to some extent. Pharmacokinetic analysis revealed that A7‐MMCD had nearly the same properties in the body as MMCDan (MMCD with an anionic charge), i.e. those of a negatively charged macromolecule. Both A7‐MMCD and MMCDan had relatively similar tissue uptake rate indices for the liver and spleen. The tumor uptake rate index for SW1116 was about 2.5 times greater than that for S180, and the total amount of 111In‐A7‐MMCD accumulated in SW1116 was calculated to be approximately 5 times greater than the amount in S180. These results indicated that A7‐MMCD could achieve site‐specific targeting in the body. Furthermore, in the therapeutic experiment using SW1116 implanted subcutaneously, A7‐MMCD suppressed tumor growth significantly, compared to free MMC and MMCDan. These results suggest that in designing an monoclonal antibody‐drug conjugate via an intermediary, the physicochemical properties of intermediate macromolecules must also be taken into consideration to obtain a high degree of efficacy in vivo.
Overexpression of c‐Myc Inhibits the Appearance of a Specific DNase I Hypersensitive Site in the β‐Globin Chromatin in Murine Erythroleukemia Cells
Authors: Takada Shinji , Obinata Masuo
Published: August 2005

To investigate the molecular mechanism of regulation of differentiation by c‐Myc, we examined the acquisition of DNase I hypersensitivity by;β‐globin chromatin in an MEL cell transformant (38–2) in which the c‐myc gene was placed under the control of a metallothionein gene promoter. Of the three DNase I hypersensitive sites around the β‐globin gene which were detected in MEL cells after DMSO treatment, the appearance of a specific DNase I hypersensitive site near the promoter was greatly reduced by the Overexpression of c‐Myc. This result suggests that c‐Myc regulates β‐globin expression by interfering with the establishment of an active chromatin conformation around the β‐globin gene.
Inhibitory Effect of 2‐0‐Octadecylascorbic Acid in Agglutination Assay with Concanavalin A; Short‐term Examination of Rat Urinary Bladder Carcinogenesis
Authors: Suzuki Mika , Wakabayashi Keiji , Sone Hideko , Kushida Hiromi , Sugiyama Kiyoshi , Kakizoe Tadao , Nagao Minako , Sugimura Takashi
Published: August 2005

A derivative of ascorbic acid, 2‐O‐octadecylascorbic acid (CV‐3611), is a strong scavenger of active oxygen species. We examined the effect of CV‐3611 on a short‐term test of bladder Carcinogenesis, using Concanavalin A (Con A) ‐dependent agglutination of isolated bladder epithelial cells. Rats were given 0.01%N ‐butyl‐N(4‐hydroxybutyl)nitrosamine (BHBN) for 1 week, and then 5% sodium saccharin or 2% DL‐tryptophan or 0.01% BHBN alone or with 0.002, 0.006 or 0.02% CV‐3611 for 3 weeks. Treatment with CV‐3611 reduced the effects of the bladder tumor promoters sodium saccharin and DL‐tryptophan by 48–86 and 65–87%, respectively. CV‐3611 also reduced the number of aggregates of bladder epithelial cells from rats treated with BHBN for 4 weeks. These results suggest that CV‐3611 has a suppressive effect on rat bladder Carcinogenesis.
Sequence‐dependent Termination of Mammalian DNA Polymerase Reaction by a New Platinum Compound, (–)‐(R)‐2‐Aminomethylpyrrolidine(1,1‐cyclobutane‐dicarboxylato)‐2‐platinum(II) Monohydrate
Authors: Iwata Mitsumasa , Izuta Shunji , Suzuki Motoshi , Kojima Kiyohide , Furuhashi Yoshihito , Tomoda Yutaka , Yoshida Shonen
Published: August 2005

We examined the mechanisms of the inhibition of DNA synthesis by a new platinum compound, (‐)‐(R)‐2‐aminomethylpyrrolidine(1,1‐cyclobutane‐dicarboxylato)‐2‐platinum(II) monohydrate (DWA‐2114R), a derivative of the antitumor drug cis‐ diamminedichloroplatinum(II) (CDDP), using prokaryotic and eukaryotic DNA polymerases. Preincubating activated DNA with CDDP or DWA‐2114R reduced its template activity for prokaryotic and eukaryotic DNA polymerases in a dose‐dependent manner. DWA2114R required six times greater drug concentration and two times longer incubation time to show the same decrease of the template activity compared to CDDP. Treatment of primed pUC118 ssDNA templates with the two drugs followed by second‐strand synthesis by prokaryotic and eukaryotic DNA polymerases revealed that DWA2114R bound to DNA in a similar manner to CDDP and these adducts blocked DNA elongation by DNA polymerases of eukaryotes as well as of prokaryotes. With these two drugs, the elongations by E. coli DNA polymerase I (Klenow fragment), T7 DNA polymerase and calf thymus DNA polymerase α were strongly arrested at guanine‐guanine sequences (GG). Stop bands were also observed at adenine‐guanine sequences (AG) guanine‐adenine‐guanine sequences (GAG) and mono‐guanine sequence (G). Calf testis DNA polymerase β was also arrested efficiently at AG, GAG and G, but much more weakly at GG. This pattern was common to DWA2114R and CDDP.
In vivo Tumor Growth Enhancement by Granulocyte Colony‐stimulating Factor
Authors: Segawa Kotaro , Ueno Yoshio , Kataoka Tateshi
Published: August 2005

The intraperitoneal administration of human recombinant granulocyte colony‐stimulating factor (G‐CSF) enhanced the growth of intradermally inoculated tumor in mice; in a Meth A fibrosarcoma model, G‐CSF administration significantly shortened the latency before tumor appearance, accelerated the increase of tumor size, shortened the survival time of tumor‐bearing mice and increased the incidence of lethal tumor growth. A similar growth‐enhancing effect of G‐CSF was observed in models employing Meth 1 fibrosarcoma, colon carcinoma 26, and L1210 leukemia, although not all the effects were statistically significant. In vitro study showed that G‐CSF did not enhance Meth A growth in suspension culture or in soft agar. These data suggest that G‐CSF enhances the Meth A growth not directly but through the mediation of host factors. The accumulation of neutrophils was histologically observed in the tumor nodule, the blood, and the spleen in mice given G‐CSF repeatedly. The spleen cells and the peripheral blood leukocytes of G‐CSF‐injected mice enhanced Meth A growth in vitro as compared with those of mice injected with physiological saline. These results suggest the possibility that the in vivo growth of tumor cells was enhanced by G‐CSF‐induced overproduction of cells including neutrophils.
T Cell Receptor‐independent Cell‐mediated Cytotoxicity by Nude Mouse Lymphokine‐activated Killer Cells
Authors: Nishimura Takashi , Togashi Yuji , Wakamiya Nobutaka , Hashimoto Yoshiyuki , Yagita Hideo , Okumura Ko , Habu Sonoko
Published: August 2005

Lymphokine‐activated killer (LAK) cells, which can lyse a variety of tumor cells, can be induced from both normal and athymic nude mouse spleen cells by culture with high doses of recombinant interleukin 2 (rIL‐2). LAK cells generated from nude mouse spleen cells (Nude‐LAK cells) express just Thy 1.2 antigen, but not CD4 and CDS antigens. Nude‐LAK cells express neither T3 molecule, T cell receptor (TCR) αβ nor TCR γδ on their cell surface. The lack of TCR expression on Nude‐LAK cells was confirmed by the results of northern blot analysis. LAK cells generated from normal mouse spleen cells (Nor‐LAK) express TCR α, β transcripts, while Nude‐LAK cells express only sterile TCR β transcript, but not TCR α transcript. TCR γδ transcripts were scarcely detected in both Nor‐LAK cells and Nude‐LAK cells. Thus, it is strongly suggested that Nude‐LAK cells can recognize and lyse tumor cells by TCR‐independent mechanisms. Monoclonal antibody against lymphocyte function‐associated antigen (LFA‐1) molecule can block the Cytotoxicity of Nude‐LAK cells, indicating an important role of such accessory molecules in Nude‐LAK cell‐mediated Cytotoxicity.
Hereditary Low Level of Plasma Ceruloplasmin in LEC Rats Associated with Spontaneous Development of Hepatitis and Liver Cancer
Authors: Ono Takao , Abe Syuiti , Yoshida Michihiro C.
Published: August 2005

Both young (5 weeks old) and old (61 100 weeks old) hereditary hepatitis LEC rats showed a markedly low level of plasma ceruloplasmin (Cp) ferroxidase activity as compared with that of age‐matched LEA and BN strain rats. This trait was genetically examined hy the use of (BN × LEC) F1 hybrid and (F1× LEC) backcross rats. The F1 hybrids never developed hepatitis and showed a similar level of Cp to that found in the parental BN rats. Among the backcross rats with about 1:1 segregation rate for hepatitis, affected rats had a remarkably decreased level of Cp, as found in LEC rats, whereas unaffected rats exhibited a similar level of Cp to that of BN, F1 and LEA rats. These results indicate that the low level of Cp is heritable in a single autosomal recessive mode in LEC rats. The observed tight link between the low Cp level and the hepatitis in LEC rats suggests that defective copper metabolism may he associated with the occurrence of hepatitis in LEC rats, since Cp is a copper‐binding protein primarily involved in copper transport from the liver.
Promotion of BALB/3T3 Cell Transformation by the Okadaic Acid Class of Tumor Promoters, Okadaic Acid and Dinophysistoxin‐1
Authors: Sakai Ayako , Fujiki Hirota
Published: August 2005

Okadaic acid and dinophysistoxin‐1 are non‐12‐O‐tetradecanoylphorbol‐13‐acetate (non‐TPA)‐type tumor promoters, which enhance chemically induced tumorigenesis on mouse skin through a different mechanism from that of TPA. In the present study, we examined the promoting effects of these okadaic acid class tumor promoters on a two‐stage transformation using BALB/3T3 cells which was designed to simulate in vivo two‐stage carcinogenesis. Cells were treated first with a low dose of the initiator 3‐methylcholanthrene (MCA) and then with a test chemical. Okadaic acid and dinophysistoxin‐1 significantly enhanced the MCA‐induced cell transformation. Okadaic acid tetramethyl ether, an inactive compound, did not affect the transformation of MCA‐treated cells. The okadaic acid class of tumor promoters failed to induce transformation without pretreatment by MCA. Okadaic acid did not show initiating activity in the two‐stage transformation assay in which cells were treated first with okadaic acid and then with TPA. These results indicate that this transformation assay with BALB/ 3T3 cells is useful to predict tumor‐promoting activity of non‐TPA‐type as well as TPA‐type tumor promoters, before long‐term in vivo two‐stage carcinogenesis experiments are carried out.
Detection and Typing of Multiple Genital Human Papillomaviruses by DNA Amplification with Consensus Primers
Authors: Yoshikawa Hiroyuki , Kawana Takashi , Kitagawa Kyoko , Mizuno Masahiko , Yoshikura Hiroshi , Iwamoto Aikichi
Published: August 2005

Many types of human papillomavirus (HPV) are associated with genital lesions. In order to develop simple and sensitive diagnostic procedures for HPV infection, we took advantage of the polymerase chain reaction (PCR). We compared the published nucleotide sequences of the LI region from six genital HPV types and designed a pair of consensus primers for LI region. The PCR with the consensus primers for LI region (Ll‐PCR) could amplify at least nine genital HPV types, 6,11, 16, 18, 31, 33, 42, 52 and 58, and the amplified HPV DNA could be typed by subsequent restriction mapping. Ll‐PCR was compared to Southern blot analysis and also to the consensus primer‐mediated PCR for E6 region (E6‐PCR) described before. Although both our PCR systems are nonradioactive, the sensitivity in detecting HPV DNA was even better than that obtained by using Southern blot analysis. By means of the PCR systems we detected HPV DNA in 100% of cervical condylomas (10/10), 92% of cervical intraepithelial neoplasias (33/36) and 96% of invasive cervical carcinomas (53/55), while we detected HPV DNA in 12% of normal cervices (12/102).
Alteration of Physiological Activity of Activated Macrophages through L‐Arginine Metabolism
Authors: Takema Morio , Inaba Kayo , Okazaki Ken‐ichi , Uno Kazuko , Tawara Katsuya , Muramatsu Shigeru
Published: August 2005

Our aim in this study was to define the effect of L‐arginine on macrophages (Mø) in relation to the decay of tumoricidal activity of activated Mø. We found that the activated M0 retained their cytotoxicity when cultured in L‐arginine‐deficient medium but not in conventional medium. Such a decline of tumoricidal activity was associated with increase of glucose consumption and concomitant lactate production, resulting in Mø death. Addition of glucose to the culture medium of activated Mø appeared to cause only a slight delay of the decrease of tumoricidal activity and Mø death. These events were also coincident with a decrease of electron transport activity in mitochondria. Cytological observation by electron microscopy clearly showed the structural alteration or destruction of mitochondria, which preceded the changes of other physiological and functional activities. These results demonstrate that the L‐arginine‐dependent cytolytic activity against tumor target cells also impairs M0 functions and ultimately induces Mø death, which is primarily mediated by the inhibition of mitochondrial activity.
Immunohistochemical Expression of Blood Group Substances and Related Carbohydrate Antigens in Breast Carcinoma
Authors: Nakagoe Tohru , Fukushtma Kiyoyasu , Hirota Masaki , Kusano Hiroyuki , Kawahara Katsunobu , Ayabe Hiroyoshi , Tomita Masao , Kamihira Shimeru
Published: August 2005

In forty–one carcinomas and sixteen benign lesions (fibroadenoma and mastopathy) of the human breast, immunohistochemical expression of sialylated and non–sialylated forms of both Lea and Lex, and the A, B, and H type 2 blood group substances were studied by using an indirect immunoperoxidase staining. In normal ductal epithelium and benign lesion of breast, Lewis–related antigens were mostly expressed. Breast carcinomas showed these antigens with the following frequencies: Lea, 31.7% (13/41); sialyl Lea, 56.1% (23/41); Lex, 46.3% (19/41); sialyl Lex, 68.3% (28/41); A/B/H type 2, 38.1% (16/41). Sialylated forms of Lea and Lex were observed more frequently than their respective non–sialylated forms in breast carcinomas. In both one normal epithelium and four carcinomas of breast with Le(a−b‐) phenotype, the expressions of type 2 antigens were observed, while type 1 antigens were not consistently expressed. Although compatible expression was observed in all specimens of both normal epithelium and benign lesion of breast, twenty–four cases with the deletion of A and/or B antigens, six cases with H type 2 accumulation and one case with incompatible expression were demonstrated in breast carcinoma. Thirty–one breast carcinomas which showed the deletion of A/B/H type 2 expressed the Lewis–related antigens more frequently than nine cases which showed compatible expression. These results suggested that the activation of terminal fucosyltransferase and sialyltransferase as well as inactivation of some glycosyltransferases had occurred in cancer cell membrane, and sialyl Le1, defined by a new monoclonal antibody CSLEX1, may be useful as a tumor–associated antigen independent of Lewis blood group type in breast cancer.
Suppressive Activity of Interleukin 4 on the Induction of Antigen‐specific Cytotoxic T Cells in Humans
Authors: Yamashita Uki , Tanaka Yoshiya
Published: August 2005

The effect of interleukin 4 (IL 4) on the induction of cytotoxic T cells (CTL) was studied by using human peripheral blood lymphocytes in vitro. IL 4 suppressed the induction of CTL specific for allogeneic antigens in a concentration‐dependent manner. However, IL 4 did not suppress proliferative responses induced with allogeneic antigens or mitogens. The suppressive effect of IL 4 on CTL induction was observed when IL 4 was added at the early period of the CTL induction culture, but not at the later period. Furthermore, IL 4 did not suppress the effector function of CTL to target cells. IL 4 suppressed the production of IL 1 by monocytes/macrophages and the production of IL 2 and the expression of IL 2 receptors on T cells. Moreover, IL 4 suppressed the induction of lymphokine‐activated killer cells. These results suggest that IL 4 has a suppressive activity on the induction of killer cells in humans.
Photodynamic Cell Killing Effects and Acute Skin Photosensitivity of Aluminum‐chloro‐tetrasulfonated Phthalocyanine and Hematoporphyrin Derivative
Authors: Komatsu Kazuto
Published: August 2005

Aluminum‐chloro‐tetrasulfonated phthalocyanine (PC) showing an absorption peak at 678 nm was compared to hematoporphyrin derivative (MpD), a photosensitizer commonly used in the photodynamic therapy (PDT) of cancers. In vitro studies: KK‐47 cells were exposed to long‐wavelength ultraviolet (UVA) or red light (>600 nm, >640 nm and >660 nm) after drug sensitization. With UVA irradiation, a higher photodynamic cell killing effect was observed in the cells treated with HpD than with PC. However, with red light irradiation (both > 640 nm and >660 nm) PC resulted in greater cell damage. PC was less toxic to KK‐47 cells in the dark. In vivo studies: Using a gold vapor laser (GVL: 627.8 nm, 200 mW/cm2, 200 J/cm2), the photodynamic tumor response was determined in C3H/He mice bearing transplantable squamous cell carcinoma. No significant difference was observed in the tumor volume between the PC and HpD groups, except that the PC group (10.0 mg/kg body weight) showed a significantly higher remission rate (3/6) than the control group (0/10, P<0.05). Skin Photosensitivity test: Skin photosensitivity was estimated by measuring changes in back skin thickness due to photosensitization. With UVA irradiation, a stronger skin reaction was observed in the HpD group, while with visible light irradiation there was no significant difference between the HpD and PC groups. Based on the superior cell killing effect with red light, reduced toxicity to the cells in the dark and mild skin reaction with UVA, PC may be a more promising photosensitizer for PDT.
A New Form of Specific Targeting Cancer Immunotherapy Using Anti‐tumor Monoclonal Antibody‐conjugated Lymphokine‐activated Killer Cells
Authors: Shiraiwa Hiroshi , Sekine Teruaki , Tobisu Ken‐ichi , Kakizoe Tadao , Koiso Kenkichi
Published: August 2005

Cross‐linking of effector T cells to target cancer cells augments their tumor lytic activities. Here we describe a new method of conjugating lymphokine‐activated killer (LAK) cells with cancer‐specific monoclonal antibody. The LAK cells were biotinylated, treated with avidin, and conjugated with biotinylated monoclonal antibody. These monoclonal antibody‐conjugated LAK cells showed specifically enhanced killing activities against anti‐tumor antibody‐reactive cancer cells, and cold target cells specifically inhibited their activities.
Work‐related Bladder Cancer Risks in Male Japanese Workers: Estimation of Attributable Fraction and Geographical Correlation Analysis
Authors: Yaraaguchi Naohito , Watanabe Shaw , Okubo Toshiteru , Takahashi Ken
Published: August 2005

One hundred and forty‐nine relative risks (RRs) on occupations and bladder cancer were extracted from 27 case‐control studies and geometric means were calculated for 27 occupations as summary RRs. Those in high risk occupations, among whom the summary RRs significantly exceeded unity, were found to be petroleum workers (RR=3.51), dye workers (RR = 3.38), machinists (RR=2.76), drivers (RR = 2,22), rubber workers (RR=2.19), printers (RR = 2.12), clothing/tailors (RR = 2,10), wood workers/carpenters (RR=1.70), miners (RR = 1.68), textile workers (RR = 1.63), mechanics (RR=1,54), engineers (RR = 1,54), leather workers (RR=1.49), painters (RR=1.48) and chemical workers (RR = 1.44). The estimated numbers of male bladder cancers in 1980 in the high risk and other occupations were 820 and 2065 cases, respectively. The estimated number of work‐related bladder cancer was 548 cases, the attributable fraction being 19% in active employees and 12% in the entire population. The geographical distribution of male bladder cancer deaths in Japan was positively correlated with the distributions of workers in the transport/communications industry and in tertiary industries including services, wholesale, retail, finance and insurance, whereas it was negatively correlated with the distribution of workers in the agricultural industry. These findings suggest that workers in certain industries may be at high risk of bladder cancer, but lifestyle modification associated with urbanization and industrialization could be an alternative explanation.
Modifying Effects of Various Chemicals on Preneoplastic and Neoplastic Lesion Development in a Wide‐spectrum Organ Carcinogenesis Model Using F344 Rats
Authors: Fukushima Shoji , Hagiwara Akihiro , Hirose Masao , Yamaguchi Shuji , Tiwawech Danai , Ito Nobuyuki
Published: August 2005

Modifying potentials of various chemicals on tumor development were investigated in a wide‐spectrum organ carcinogenesis model using male F344/DuCrj rats. The animals were treated with N‐nitroso‐diethylamine (100 mg/kg body weight, ip, single injection at the commencement of the study), N‐methyl‐N‐nitrosourea (20 mg/kg body weight, ip, 4 times during weeks 1 and 2) and N‐bis(2‐hydroxypropyl)nitrosamine (0.1% in drinking water, during weeks 3 and 4) for multi‐organ initiation and then were given one of 14 test chemicals including 6 hepatocarcinogens, 7 non‐hepatocarcinogens and 1 non‐carcinogen, or basal diet for 16 weeks. All rats were killed at the end of week 20, and the major organs were carefully examined for preneoplastic and neoplastic lesions. Immunohistochemical demonstration of glutathione S‐transferase‐positivc foci was also used for quantitative assessment of liver preneoplastic lesion development. Modifying effects were shown for 11 out of 14 test agents in the liver, forestomach, glandular stomach, lung, urinary bladder or thyroid, 7 of them targeting more than two organs. This was the first demonstration to our knowledge that cloflbrate possesses enhancing potential for urinary bladder carcinogenesis and an inhibiting effect on thyroid carcinogenesis. Caprolactam showed no effect in any organ, in agreement with its established inactivity. The results indicated that the system could be reliably applied as a medium‐term multiple organ bioassay for assessment of the modification potential of test agents in unknown target sites.
Time‐ and Dose‐dependent Induction of Invasive Urinary Bladder Cancers by N‐Ethyl‐N‐(4‐hydroxybutyl)nitrosamine in B6C3F1 Mice
Authors: Tamano Seiko , Hagiwara Akihiro , Suzuki Emako , Okada Masashi , Shirai Tomoyuki , Fukushima Shoji
Published: August 2005

A sequential investigation of N‐ethyl‐N‐(4‐hydroxybutyl)nitrosamine (EHBN) bladder carcino‐genesis was performed in male B6C3F1mice maintained ad libitum on tap water containing 0.025% EHBN for 4, 12, 20, 28 and 36 weeks. A total of 81 invasive tumors, comprising 55 smiamous cell carcinomas (SCCs) (68%), 25 transitional cell carcinomas (TCCs) (31%) and 1 adenocarcinoma (1%) were found. Of these, 23 (22 SCCs and 1 TCC) demonstrated invasion to the prostate, 3 metastasized to the lung, and 2 spread by peritoneal seeding. The anaplastic grade and extent of invasion of the SCCs significantly exceeded those of the TCCs. The results suggested a histogenetic pathway from simple dysplasia through papillary or nodular dysplasia and/or carcinoma in situ to eventual development of invasive carcinomas
Sex Differences in o‐Phenylphenol and Sodium o‐Phenylphenate Rat Urinary Bladder Carcinogenesis: Urinary Metabolites and Electrolytes under Conditions of Aciduria and Alkalinuria
Authors: Hasegawa Ryohei , Fukuoka Masamichi , Takahashi Terue , Yamamoto Atsushi , Yamaguchi Shuji , Shibata Masa‐Aki , Tanaka Akira , Fukushima Shoji
Published: August 2005

F344 male and female rats were administered 1.25%o‐phenylphenol (OPP) or 2% sodium o‐ phenylphenate (Na‐OPP) in combination with 3% NaHCO3 or 1% NI4Cl for 8 weeks and changes in the urinary bladder histopathology and the urinary components were examined. Administration of OPP with NaHCO3 resulted in marked urothelial liyperplasia in the urinary bladder of male rats, the response being less pronounced in females. OPP alone exerted no proliferative effect and NaHCO3 induced only slight hyperplasia in males. Na‐OPP alone induced mild hyperplastic lesions only in males, this being completely prevented by concomitant administration of NH4C1. The findings thus demonstrated a clear correlation between hyperplastic response and reported carcinogenic potential of these treatments. Of the urinary factors examined, increases in levels of pH and sodium ion concentration were positively associated with proliferative lesions especially in males, although the findings failed to explain the sex difference. Urinary concentrations of non‐conjugated forms of OPP metabolites were also not directly correlated with the development of hyperplasias. Thus, changes in individual urinary factors presumably affect nrothelial proliferation in combination rather than separately. The presence of OPP metabolites, including 2‐phenyl‐l,4‐benzoquinone, in the urine may be unimportant in the OPP urinary carcinogenesis even under conditions of alkalinuria and high sodium ion concentration.
Cytosolic‐nuclear Tumor Promoter‐specific Binding Protein: Association with the 90 kDa Heat Shock Protein and Translocation into Nuclei by Treatment with 12‐O‐Tetradecanoylphorbol 13‐Acetate
Authors: Hashimoto Yuichi , Shudo Koichi
Published: August 2005

Suspension‐cultured HeLa cells possess a cytosolic‐nuclear tumor promoter‐specific binding protein (CN‐TPBP) which lacks protein kinase C activity. This CN‐TPBP existed in cytosol of HeLa cells, but translocated into nuclear fraction of the cells after treatment of the cells with 12‐O‐tetradecanoyl‐phorbol 13‐acetate (TPA). The translation of CN‐TPBP induced by TPA became apparent within 10 min after the treatment with TPA, and was completed within 3 h. CN‐TPBP bound TPA with the association constant of 1.4X1010M−1, and also bound teleocidin B, debromoaplysiatoxin, and thapsigargin in a mutually competitive manner. The binding affinity order of synthetic analogs of teleocidin B correlated with the adhesion‐inducing potency order of the compounds toward human leukemia cell line HL‐60. The apparent molecular weight of CN‐TPBP under non‐denaturing conditions was estimated to be 66–68 kDa. CN‐TPBP forms a complex with the 90 kDa heat shock protein, and the complex was stabilized by the presence of molybdate. These characteristics of CN‐TPBP are similar to those of the nuclear receptors of glucocorticoid and dioxin. These findings suggested that CN‐TPBP acts as a nuclear receptor for tumor promoters, and that tumor promoters may exert their biological effects by binding to CN‐TPBP.
Rapid Growth and Spontaneous Metastasis of Human Germinal Tumors Ectopically Transplanted into Scid (Severe Combined Immunodeficiency) and Scid‐nudestreaker Mice
Authors: Nomura Taisei , Takahama Yousuke , Hongyo Tadashi , Takatera Hiroshi , Inohara Hidenori , Fukushima Hisao , Ono Shiro , Hamaoka Toshiyuki
Published: August 2005

Xenograft acceptance, growth and spontaneous metastasis of ectopically transplanted human germinal tumors were compared among scid mice, athymic nude mice and F2 hybrids constructed from scid and nude mice, in relation to the impairments of T and B cell functions in these mice. In scid mice which are deficient in T and B cell functions, human yolk sac tumor (YST‐2) that originated from the ovary grew to enormous sizes in 100% of the animals after both subcutaneous and intraperitoneal transplantation, while only half (59.1% and 51.9%) of the subcutaneous and none of the intraperitoneal transplants were accepted in usual athymic nude mice (BALB/c‐nu/nu and CDl‐nu/nu). The YST‐2 grew rapidly in scid mice, developing 3 to 10 times larger tumors compared to nude‐streaker (AKR/J‐nustr/nustr) and usual nude mice, respectively. Furthermore, ectopically transplanted tumors spontaneously metastasized to distant organs (mostly to the lung) in scid mice (but less frequently in leaky scid mice), while metastases have never been found in nude mice. Although a xenograft of human classic (typical or pure) seminoma of the testis has never been established in nude mice, it grows slowly in one‐third (36.4%) of scid mice and very rapidly in all of scid‐nu (scid/scid; nustr/nustr) double mutant mice. Spontaneous metastases of xenografted seminomas were also observed in distant organs (lymph node, lung, liver, spleen, and kidney). The metastastic distribution of the two human germinal tumors in scid and scid‐nustr mice mimics that found in human. These results (xenograft acceptance, growth of transplanted tumors and degree of metastatic spread) were compatible with the level of T and B cell impairments indicated by FACS analysis, as well as mitogen responses, serum IgG and morphological features of the thymus.
Allele Loss on Chromosome 11 in a Pituitary Tumor from a Patient with Multiple Endocrine Neoplasia Type 1
Authors: Yoshimoto Katsuhiko , Iwahana Hiroyuki , Kubo Katsuyuki , Saito Shiro , Itakura Mitsuo
Published: August 2005

We have examined the allele loss of chromosome 11 in a pituitary tumor from a patient with familial multiple endocrine neoplasia type 1 (MEN 1). The extensive loss of chromosome 11, including loci of D11S149, HRAS1 and F2, was detected by the loss of heterozygosity. All of the lost alleles of these loci were transmitted from the unaffected father and not from an affected mother. This is the first evidence of allele loss of chromosome 11 in a pituitary tumor of MEN 1 and supports the idea that similar allelic deletion of MEN1 locus on chromosome 11 is the common genetic basis for tumorigenesis in the pituitary, endocrine pancreas, and parathyroid gland in MEN 1.
Co‐production of Interleukin‐1 and Interleukin‐6 in Tumor Cell Lines Elaborating Colony‐stimulating Factors
Authors: Okuno Yoshiaki , Takahashi Takayuki , Suzuki Akira , Fukumoto Manabu , Nakamura Kishiko , Imura Hiroo
Published: August 2005

Six carcinoma cell lines elaborating colony‐stimulating factors (CSFs) were examined by enzyme‐linked immtmosorbent assay and Northern blotting to determine whether or not they co‐produced interleukin‐lα (IL‐lα), IL‐lβ and IL‐6. All 6 cell lines were co‐producers; IL‐lα was produced in all 6, IL‐6 in 5 and IL‐1β in 3 of them. These results indicate that IL‐1 and IL‐6 are commonly co‐produced in CSF‐producing tumors.
Establishment and Characterization of a New Spontaneous Metastasis Model of Human Gastric Carcinoma in Nude Mice
Authors: Nakanishi Hayao , Yasui Kenzo , Yamagata Sadako , Shimizu Satoru , Ando Shoji , Hosoda Syun
Published: August 2005

A poorly differentiated medullary carcinoma of human stomach, designated HY–1, was successfully transplanted to nude mice by either the subcutaneous or intramuscular route for five generations. The transplanted tumor showed spontaneous lung metastases in nearly 100% of KSN and Balb/c female nude mice. There were over 20 visible lung metastatic nodules in KSN and Balb/c nude mice bearing tumors for over SO days. Immunostaining of type IV collagen and electron microscopy revealed that tumor cells were often in direct contact with basement membrane (BM) of tumor blood vessels in the primary tumor tissue. At the site of contact between tumor cells and vascular BM, focal disappearance of the BM, disruption of endothelial cells and entry of tumor cell clusters into vascular lumen were observed. Immunostaining of 72 kDa gelatinase/type IV collagenase demonstrated that tumor cells expressed this enzyme in their cytoplasm. These results suggest that spontaneous metastasis of this tumor may be partly due to a marked tendency to vascular invasion involving the following sequential events: tumor cell contact with vascular BM, BM degradation possibly by 72 kDa gelatinases and endothelial disruption. This model could be a useful tool for understanding the mechanism of hematogenous metastasis of human gastric cancer.
A Case‐Control Study of Stomach Cancer and Its Genesis in Relation to Alcohol Consumption, Smoking, and Familial Cancer History
Authors: Tominaga Keigo , Koyama Yasuo , Sasagawa Michizou , Hiroki Masae , Nagai Masaki
Published: August 2005

A case‐control study of stomach cancer and its genesis has been conducted in relation to alcohol consumption, cigarette smoking, and familial cancer history. Two hundred and ninety‐four cancer cases, discovered by mass screening and histologically verified after endoscopic examination, have been compared with 588 randomly selected controls, who received the same early detection program and were verified as being free of the disease. No statistically significant association was observed between the development of stomach cancer and alcohol consumption or familial cancer history. However, the development of stomach cancer was found to have a positive correlation with smoking (relative risk for those who smoke less than 19 cigarettes/day, 3.56: 95% confidence interval, 2.39 to 5.31; relative risk for those who smoke more than 20 cigarettes/day, 2.58: 95% confidence interval, 1.60 to 4.17). The results of this study suggest that cigarette smoking appears to have a more harmful effect on the development of stomach cancer than either alcohol consumption or a familial history of cancer. The high relative risk of smoking revealed by this study implies that further research on the effects of smoking in the development of stomach cancer would be desirable.
Inhibition by Xiao‐chai‐hu‐tang (TJ‐9) of Development of Hepatic Foci Induced by N‐Nitrosomorpholine in Sprague‐Dawley Rats
Authors: Tatsuta Masaharu , Iishi Hiroyasu , Baba Miyako , Nakaizumi Akihiko , Uehara Hiroyuki
Published: August 2005

The effect of Xiao‐chai‐hu‐tang (TJ‐9) on hepatocarcinogenesis induced by N‐nitrosomorpholine (NNM) was investigated in male Sprague‐Dawley rats. Rats were given normal chow pellets containing 0.5% or 1.0% TJ‐9 until the end of the experiment, and drinking water containing NNM for 8 weeks. Pre‐neoplastic and neoplastic lesions staining for γ‐glutamyl transpeptidase (GGT) or the placenta] type of glutathione‐S‐transferase (GST‐P) were examined histochemically. In Week 15, quantitative histological analysis showed that prolonged treatment with 0.5% TJ‐9 significantly reduced the number and volume of GGT‐positive and GST‐P‐positive hepatic lesions. Treatment with 1.0% TJ‐9 inhibited the development of GGT‐positive and GST‐P‐positive lesions, but was less effective than 0.5% TJ‐9. Administration of 0.5% TJ‐9 also caused a significant increase in the proportion of helper T lymphocytes and a significant decrease in the labeling index of pre‐neoplastic lesions. These findings indicate that TJ‐9 inhibits the development of hepatic foci.
Localization of Acidic and Basic Fibroblast Growth Factor mRNA in Human Brain Tumors
Authors: Akutsu Yasuhiko , Aida Takeo , Nakazawa Shozo , Asano Goro
Published: August 2005

Acidic and basic fibroblast growth factors (aFGF and bFGF) are closely related peptide mitogens acting on both mesoderm‐ and neuroectoderm‐derived cells, including fibroblasts, endothelial cells and glial cells. In order to identify the expression of mRNAs for these growth factors, in situ hybridization using human aFGF and bFGF RNA probes was performed in 24 human brain tumors. The mRNAs for aFGF and bFGF were expressed in the cells of various tumors (1/1 and 1/1 astrocytoma, 2/2 and 2/2 anaplastic astrocytomas, 6/6 and 6/6 glioblastomas, 4/4 and 4/4 meningiomas, 3/3 and 3/3 schwannomas, 1/2 and 1/2 pituitary adenomas, 4/4 and 4/4 metastatic carcinomas, 0/1 and 0/1 hemangioblastoma, 0/1 and 0/1 craniopharyngioma) and were also detected in endothelial cells and surrounding neuronal cells of brain tumors. These results suggest the possibilities that aFGF and bFGF contribute to the uncontrolled growth of tumor cells and the proliferation of endothelial cells in autocrine and paracrine manners, and that the expression of mRNAs for these growth factors in the surrounding neuronal cells results in enhancement of tumor growth.
Augmentation and Suppression of Release of Tumor Necrosis Factor from Macrophages by Negatively Charged Phospholipids
Authors: Yui Satoru , Yamazaki Masatoshi
Published: August 2005

We recently reported that some lipid species of cell membranes and lipoproteins induced the growth of peripheral macrophages. In this study, the effects of phospholipids on tumor necrosis factor (TNF)‐releasing activity of macrophages were examined. Ten to 20 μg/ml of cardiolipin, which is a suboptimal concentration for macrophage growth‐stimulation, augmented macrophage TNF release triggered by lipopolysaccharide (LPS) in vitro. This priming effect appeared with 1 day of preincubation and was still potent on day 3, whereas the priming effect of interferon‐γ (IFN‐γ) peaked at 3 h and then gradually decreased. In contrast, a high concentration of cardiolipin (40 μg/ml) which is optimal for the induction of macrophage growth, completely suppressed LPS‐triggered TNF release from not only untreated macrophages but also IFN‐γ‐primed macrophages. The suppressive effect was potent even with 3 h preincubation, was still potent on day 3, and was not abolished by indomethacin. Cardiolipin had scarcely any effect on the triggering activity of LPS. Similar augmentative and suppressive activities were observed in peroxidized phosphatidylserine, which is also highly active in inducing macrophage growth, but was not found in native phosphatidylserine, which is less active, nor in phosphatidylcholine, which is an inactive species toward macrophage growth. These results suggest that lipids may be important endogenous factors in regulating both activation and growth states of peripheral macrophages.