bluuebunny/crossref_metadata_2025 huggingface dataset:
Distribution of Transforming Growth Factor‐β and Its Receptors in Gastric Carcinoma Tissue
Authors: Kai Toshihiro , Taketazu Fumitoshi , Kawakami Masanobu , Shimanuki Kimiyoshi , Yamada Shigeki , Miyazono Kohei , Kato Mitsuyasu , Miyata Michio
Published: August 2005

The distribution of the three mammalian isoforms of transforming growth factor (TGF)‐β (TGF‐β1,‐β2, and ‐β3) as well as their signaling receptors, TGF‐β type I and type II receptors (TβR‐I and TβR‐II, respectively), in gastric carcinoma tissue was examined by immunohistochemistry using specific antibodies. Tissue specimens were obtained from 25 cases of gastric carcinoma, which were classified into two groups according to Lauren's classification, i.e. 15 cases of diffuse carcinoma and 10 cases of intestinal carcinoma. In normal gastric mucosa apart from carcinoma nests, all of TGF‐β1, ‐β2, ‐β3, TβR‐I and TβR‐II were clearly demonstrated in fundic glands. In sharp contrast, none of them was detectable in surface mucous cells. In carcinoma cells, strong staining for TGF‐β1, ‐β2 and β3 was obtained only in diffuse‐type carcinoma. In particular, carcinoma cells scattered as single cells or small nests had a tendency to show strong staining for TGF‐βs. The receptors tended to be distributed concomitantly with the ligands, and diffuse‐type carcinoma showed stronger receptor staining than intestinal‐type carcinoma. In cancer stroma, TGF‐βs and receptors were detected in both diffuse and intestinal types, but the area with positive staining was wider and more dispersed in diffuse‐type carcinoma than in intestinal carcinoma. These results suggest that TGF‐β may contribute in part to the variety of histogenesis and mode of progression of gastric carcinoma.
A Quantitative Trait Locus in Major Histocompatibility Complex Determining Latent Period of Mouse Lymphomas
Authors: Kamoto Toshiyuki , Shisa Hayase , Pataer Abujiang , Lu Ling‐min , Yoshida Osamu , Yamada Yoshihiro , Hiai Hiroshi
Published: August 2005

The effects of two host genes on retro virus‐induced murine lymphoma were evaluated by studying 114 F2 intercross mice between SL/Kh and AKR/Ms mice. Out of 47 T‐lymphoma‐bearing F2 mice, 45 had the AKR‐derived dominant allele at Tlsm‐1. The length of the lymphoma latent period was not related to type of tumor. Instead, it was significantly shortened by a recessive SL/Kh‐derived allele at a major histocompatibility complex (MHC)‐linked locus on Chr. 17. A quantitative trait analysis of the latent period yielded a maximal logarithm of likelihood ratio for linkage (LOD) score of 7.06 at a class II gene within MHC. The SL/Kh‐derived recessive gene was named lla (lymphoma latency acceleration).
Improvement of Intraperitoneal Chemotherapy for Rat Ovarian Cancer Using Cisplatin‐containing Microspheres
Authors: Kumagai Seisuke , Sugiyama Toru , Nishida Takashi , Ushijima Kimio , Yakushiji Michiaki
Published: August 2005

Microspheres consisting of L‐lactic acid and glycolic acid copolymer containing cisplatin (CDDP‐PLGA) were developed to improve the delivery of cisplatin. We evaluated the effects of intraperitoneal administration of cisplatin prepared as CDDP‐PLGA in rats with ovarian cancer. The toxicity, platinum distribution, and therapeutic effects of CDDP‐PLGA were evaluated as compared with those in the case of cisplatin aqueous solution. The LD50 of CDDP‐PLGA was almost four‐fold higher than that of cisplatin aqueous solution. CDDP‐PLGA released cisplatin slowly and achieved a higher concentration in the peritoneal cavity and in peritoneal tumors for prolonged periods, while the tissue concentration of cisplatin was reduced elsewhere in the body, as compared with the case of cisplatin aqueous solution. The survival of rats with peritoneal carcinomatosis was increased by this delivery system relative to cisplatin aqueous solution. CDDP‐PLGA thus allows a higher dose to be given without increasing systemic toxicity, enhancing the therapeutic effect of cisplatin.
Gastric Tumorigenicity of 1,2‐Dimethylhydrazine on the Background of Gastric Intestinal Metaplasia Induced by X‐Irradiation in CD (SD) Rats
Authors: Ando Yasumi , Watanabe Hiromitsu , Tatematsu Masae , Hirano Kazuyuki , Furihata Chie , Fujimoto Nariaki , Toge Tetsuya , Ito Akihiro
Published: August 2005

Five‐week‐old male CD (SD) rats were X‐irradiated with a total of 20 Gy in 2 equal fractions with a 3‐day interval. After the second irradiation, rats were fed normal diet supplemented with 1% sodium chloride, which is known to increase intestinal metaplasia. 1,2‐Dimethylhydrazine (DMH) solution was injected i.m. into the back musculature at a dose of 20 mg/kg body weight weekly for 10 weeks, beginning 20 weeks after the final irradiation. Twelve months after the initial carcinogen treatment, gastric tumors in the glandular stomach were observed in 2 (3 lesions) of 30 animals in the X‐irradiated and DMH‐treated group fed diet supplemented with 1% sodium chloride. No gastric tumors were observed in the group which excluded X‐irradiation from the experimental protocol.
Detection of Low‐level Expression of P‐Glycoprotein in ACHN Renal Adenocarcinoma Cells
Authors: Kawamoto Shogo , Deguchi Takashi , Nezasa Shinichi , Yamada Shinichiro , Okano Manabu , Kawada Yukimichi
Published: August 2005

A highly sensitive reverse transcriptase‐polymerase chain reaction (RT‐PCR) assay and a flow cytometric assay were used to examine ACHN cells for the expression of P‐glycoprotein. The expression of P‐glycoprotein was detected at the RNA and protein levels in ACHN cells by RT‐PCR and flow cytometry, respectively. However, it was below the limit of detection by immunoblotting. The intracellular accumulation of adriamycin in ACHN cells was enhanced by verapamil, cyclosporin A and medroxyprogesterone acetate. Therefore, this study has demonstrated that low‐level expression of P‐glycoprotein detectable only by RT‐PCR and flow cytometry plays a significant role in reducing the intracellular concentration of antitumor agents and thus contributes to the multidrug‐resistant phenotype of ACHN cells.
Anti‐ganglioside GM2 Monoclonal Antibody‐dependent Killing of Human Lung Cancer Cells by Lymphocytes and Monocytes
Authors: Hanibuchi Masaki , Yano Seiji , Nishioka Yasuhiko , Yanagawa Hiroaki , Sone Saburo
Published: August 2005

Ganglioside GM2 (GM2) frequently appears on the cell surface of human cancers of neuroendocrine origin. A mouse‐human chimeric monoclonal antibody (mAb), KM966, against GM2 was previously found to promote the lysis of various cancer cells by human blood mononnclear cells (MNC). In this study, we analyzed the effector cells responsible for the chimeric mAb‐dependent cell‐mediated cytotoxicity (ADCC) against small cell lung cancer (SCLC) cells and examined the enhancing effect of various cytokines on the ADCC activity. The ADCC activity was assessed by 4‐h 51Cr release assay. Highly purified lymphocytes (>99%) and monocytes (>90%) were separated by centrifugal elutriation from peripheral blood MNC of the same healthy donor. KM966 induced lysis of SCLC cells mediated by both lymphocytes and monocytes to similar extents, in a dose‐dependent manner. Pretreatment of lymphocytes with various cytokines [interleukin (IL)‐2, IL‐12 and interferon‐γ] and that of monocytes with macrophage‐colony‐stimulating factor significantly augmented the killer activity against SCLC cells in the presence of KM966 mAb. KM966 was also effective for the lysis of non‐small cell lung cancer cells in direct proportion to the GM2 expression levels. These findings suggest that combined treatment of KM966 mAb with cytokines may be therapeutically useful for in vivo killing of lung cancer cells expressing GM2 through the ADCC reaction.
Biochemical Characterization of α‐Fetoprotein and Other Serum Proteins Produced by a Uterine Endometrial Adenocarcinoma
Authors: Koyama Yoshikazu , Taketa Kazuhisa , Azuma Masaki , Yamamoto Ritsu , Fujimoto Seiichiro , Nishi Shinzo
Published: August 2005

A high serum α‐fetoprotein (AFP) level was found in a patient with endometrial adenocarcinoma of the uterus, which appeared to be hepatoid on histological examination. The AFP of this unusual patient was purified hy immunoaffinity chromatography and characterized. The electrophoretic profiles on sodium dodecyl sulfate‐polyacrylamide gel electrophoresis both before and after glycopeptidase F treatment were indistinguishable from those of a hepatoma AFP. This indicates that the patient's AFP was also composed of a single polypeptide chain of Mr 67,000 and an N‐linked sugar chain of Mr 3,000. Amino acid sequence analyses of this AFP, and of AFP from hepatoma and umbilical cord serum indicated that the N‐terminal sequences were essentially the same. The sequence, Arg‐Thr‐Leu‐His‐Arg‐Asn‐Glu‐Tyr‐Gly‐Ile, was slightly different from previous reports, but matched that deduced from the cDNA sequence. AFP isoforms due to microheterogeneity of the sugar chain were analyzed by lectin affinity electrophoresis using a series of lectins. The AFP isoform profiles were distinct from those of proteins derived from cord serum, hepatoma, yolk sac tumor and gastric cancer. The reverse‐transcription of RNA from the tumor tissue followed by a polymerase chain reaction using primers with AFP‐specific sequences gave a product of the size and nucleotide sequence expected for AFP. mRNAs possessing the requisite sequences for albumin and transferrin syntheses were also detected in the tumor. The expression of these hepatocyte‐specific proteins supported the hepatoid nature of this tumor.
Detection of Gastric Cancer Micrometastases in Lymph Nodes by Amplification of Keratin 19 mRNA with Reverse Transcriptase‐Polymerase Chain Reaction
Authors: Noguchi Sbinzaburo , Hiratsuka Masahiro , Furukawa Hiroshi , Aihara Tomohiko , Kasugai Tsutomu , Tamura Sumihito , Imaoka Shingi , Koyama Hiroki , Iwanaga Takeshi
Published: August 2005

A sensitive method for the detection of gastric cancer micrometastases in lymph nodes was developed. The method was based on amplification of keratin 19 mRNA by reverse transcriptase‐polymerase chain reaction (RT‐PCR). Keratin 19 RT‐PCR showed that keratin 19 mRNA was expressed in all 12 gastric cancers, but not in any of 20 normal control lymph nodes, indicating that keratin 19 mRNA is a good target of RT‐PCR for the detection of gastric cancer micrometastases in lymph nodes. Serial dilution studies of RNA extracted from gastric cancers against RNA extracted from control lymph nodes demonstrated that the detection sensitivity of the keratin 19 RT‐PCR method was one cancer cell in 103‐105 lymph node cells. Detectability of lymph node metastases was compared between keratin 19 RT‐PCR and conventional histological examination, using 100 lymph nodes obtained from 12 gastric cancer patients. Keratin 19 mRNA was detected in all of the seven lymph nodes which were historically metastasis‐positive. Of the 93 lymph nodes which were histologically metastasisnegative, 79 were found not to express keratin 19 mRNA but 14 were found to express keratin 19 mRNA, indicating that these lymph nodes contained micrometastases which could not be detected by histological examination. These results demonstrate that keratin 19 RT‐PCR is a more sensitive method than histological examination for the detection of gastric micrometastases in lymph nodes.
Therapeutic Drug Monitoring in 21‐Day Oral Etoposide Treatment for Lung Cancer
Authors: Ando Yuichi , Minami Hironobu , Saka Hideo , Ando Masahiko , Sakai Shuzo , Shimokata Kaoru
Published: August 2005

We aimed to determine whether or not therapeutic drug monitoring is applicable to 21‐day oral etoposide treatment for lung cancer. As the starting dose, a 25‐mg capsule of etoposide was taken orally three times daily (75 mg/body). To achieve the target concentration range of 1.0 to 1.5 μg/ml, the dose was changed to two (50 mg/body) or four (100 mg/body) times a day from day 5, depending on the mean concentration obtained on days 3 and 4 (Cbefore). The mean concentration was calculated by use of a limited sampling model we constructed previously. Among 26 courses in 15 patients, two patients experienced grade 4 leukopenia plus neutropenia, and one of them died on day 20. Because nausea/emesis prevented the planned dose escalation in one patient, we excluded two courses of this patient from the pharmacokinetic analysis of dose modification. Among 5 courses with dose reduction, the Cbefore of 1.7±0.1 (μg/ml, mean±SE) was decreased to 1.3±0.2 after day 5 (Cafter). Among 7 courses with dose escalation, the Cbefore of 0.9±0.0 was increased to the Cafter of 1.2±0.1. Among the remaining 12 courses without dose modification, the Cbefore and the Cafter were 1.2±0.0 and 1.3±0.1, respectively. Hematologic toxicities tended to correlate with the drug concentration. TDM is thus applicable to oral etoposide given according to this schedule, and a larger study is now needed to confirm that the therapeutic efficacy is improved by introducing TDM.
Prevention by 2‐Mercaptoethane Sulfonate and N‐Acetylcysteine of Renal Oxidative Damage in Rats Treated with Ferric Nitrilotriacetate
Authors: Umemura Takashi , Hasegawa Ryuichi , Sai‐Kato Kimie , Nishikawa Akiyoshi , Furukawa Fumio , Toyokuni Shinya , Uchida Koji , Inoue Tohru , Kurokawa Yuji
Published: August 2005

Ferric nitrilotriacetate (Fe‐NTA) is a renal toxicant and carcinogen in rats and mice. We found that its administration results in formation of 4‐hydroxy‐2‐nonenal (HNE) in the renal proximal tubule cells of rats, and 8‐hydroxydeoxyguanosine (8‐OHdG) adducts in their DNA, suggesting a role for oxidative stress. Since 2‐mercaptoethane sulfonate (MESNA) and N‐acetylcysteine (NAC), administered orally, have been shown to increase the kidney levels of free thiol groups, their influence on the renal toxicity and carcinogenicity induced by Fe‐NTA was examined in the present study. Male Wistar rats were intraperitoneally injected with Fe‐NTA (12 mg Fe/kg), and MESNA (100 mg/kg) or NAC (200 mg/kg) was given orally 1 h before and 1 h after this treatment. The animals were killed for tissue analyses 3 h after the Fe‐NTA exposure. In accord with our previous reports, HNE‐modified protein was detected in the proximal tubules of Fe‐NTA‐treated rats by means of immunohistochemistry. Likewise, levels of 8‐OHdG in the renal nuclear DNA, lipid peroxides as thiobarbituric acid‐reactive substances in the kidneys, and blood urea nitrogen and creatinine in the serum were significantly increased by the Fe‐NTA treatment. All of these changes were completely inhibited by oral administration of MESNA or NAC. These results suggest that both of these compounds can prevent the oxidative stress induced by Fe‐NTA.
Evaluation of Angiogenic Inhibitors with an in vivo Quantitative Angiogenesis Method Using Agarose Microencapsulation and Mouse Hemoglobin Enzyme‐linked Immunosorbent Assay
Authors: Okada Naoki , Fushimi Mariko , Nagata Yuriko , Fukunaga Tomoko , Tsutsumi Yasuo , Nakagawa Shinsaku , Mayumi Tadanori
Published: August 2005

In the present work, using a previously reported in vivo quantitative tumor‐angiogenesis model, we attempted to ascertain whether this animal model is suitable for practical use in monitoring inhibitors of tumor angiogenesis. Mouse sarcoma‐180 cells, human A431 cells or rat C6 cells microencapsulated in agarose beads were implanted s.c. into C57BL/6 mice. The level of blood vessel induction at the agarose pellet site was evaluated using mouse hemoglobin enzyme‐linked immunosorbent assay on day 10 after implantation. Hydrocortisone, tetrahydro‐S, medroxyprogesterone acetate, pentosan polysulfate and suramin inhibited blood vessel growth in our in vivo tumor‐angiogenesis assay system, and heparin enhanced the antiangiogenic effects of hydrocortisone and tetrahydro‐S. These results are almost entirely consistent with those observed in common assay systems, and suggest that this method may be useful for the identification and quantitative evaluation of inhibitors of tumor angiogenesis.
The Cytocidal Activity of OK‐432‐activated Mononuclear Cells against Human Glioma Cells is Partly Mediated through the Fas Ligand/Fas System
Authors: Toda Keisuke , Shiraishi Tetsuya , Hirotsu Tatsumi , Fukuyama Kouzou , Mineta Toshihiro , Kawaguchi Shojiro , Tabuchi Kazuo
Published: August 2005

We have been applying an adoptive immunotherapy protocol to patients with malignant brain tumors using OK‐432‐activated peripheral blood mononuclear cells (OK‐MCs). In order to elucidate the mechanism of OK‐MCs' cytotoxicity, we examined the expression of Fas ligand mRNA in OK‐MCs and the cytocidal activity of these cells against a human glioma cell line, T98G which expresses a high level of Fas. The expression of Fas ligand mRNA was low in non‐treated peripheral blood mononuclear cells and was elevated by treatment with OK‐432, irrespective of the dose employed. Apoptosis of T98G cells induced by OK‐MCs was unequivocally inhibited by the pretreatment of T98G cells with ZB4 monoclonal antibody, which binds to Fas and blocks the binding of Fas ligand to Fas. These data indicate that the cytotoxic activity of OK‐MCs via apoptosis seems to be at least partly mediated by the Fas ligand/Fas system. Adoptive immunotherapy using the Fas ligand/Fas system could be a new treatment modality for human malignant brain tumors.
Failure to Detect Mutations in the Retinoblastoma Protein‐binding Domain of the Transcription Factor E2F‐1 in Human Cancers
Authors: Nakamura Takao , Monden Yoshiaki , Kawashima Kazuko , Naruke Tsuguo , Nishimura Susumu
Published: August 2005

The functions of the transcription factor E2F‐1 are regulated by the RB protein through the RB‐binding domain of E2F‐1 and this factor is considered to be an important molecule that functions downstream of the RB protein. In order to determine whether E2F‐1 that cannot bind to RB might be associated with various human cancers, we searched for mutations in the RB‐binding domain of E2F‐1 using samples of DNA from various clinical specimens obtained from 406 cancer patients (with lung, pancreatic, stomach, colon, esophageal, and hepatic cancers) by analysis of polymerase chain reaction‐mediated single‐strand conformational polymorphism. No mutations or deletions were detected in genes for E2F‐1 from any of the tumor tissues examined. These results suggest that a mutation or deletion in E2F‐1 that might affect binding of the RB protein is not involved in human cancers.
Therapeutic Drug Monitoring of Etoposide in a 14‐Day Infusion for Non‐small‐cell Lung Cancer
Authors: Ando Yuichi , Minami Hironobu , Saka Hideo , Ando Masahiko , Sakai Shuzo , Shimokata Kaoru
Published: August 2005

We investigated whether a constant plasma concentration could be obtained by the individualized administration of low‐dose, prolonged‐infusional etoposide. Etoposide was infused for 14 days at 40 mg/m2day initially in patients with inoperable non‐small‐cell lung cancer. The infusion rate was modified based upon the etoposide concentration at 24 h following the initiation of the infusion (C24) to achieve a target concentration of 1.5 μg/ml. We postulated that severe toxicities could be avoided by maintaining the steady‐state concentration at less than 2 7mu;g/ml, while antitumor activity could be expected if the steady‐state concentration was maintained at more than 1 μg/ml. In a total of 21 courses in 12 patients, the mean etoposide dose was 35±6 mg/m2 daily. The C24 was 1.8±0.4 μg/ml and ranged from 1.1 to 2.9 μg/ml. Following dose modification, the mean concentration from 96 to 336 h (Cmean) was 1.6±0.2 μg/ml and ranged from 1.2 to 2.0 μg/ml. The toxicities were well‐tolerated except for one patient with WHO grade 4 leukopenia and neutropenia who developed infectious complications. There were no treatment‐related deaths. Following dose modification, the inter‐patient variability was decreased successfully. Although this pharmacologically‐guided method needs to be validated using more patients, it could be used for therapeutic drug monitoring.
Reduced Activity of Anabolizing Enzymes in 5‐Fluorouracil‐resistant Human Stomach Cancer Cells
Authors: Inaba Makoto , Mitsuhashi Junko , Sawada Hiroko , Miike Naoko , Naoe Yoshinori , Daimon Aiko , Koizumi Kazuhisa , Tsujimoto Hiroaki , Fukushima Masakazu
Published: August 2005

The mechanism of resistance to 5‐fluorouracil (5‐FU) was studied with NUGC‐3/5FU/L, a human stomach cancer cell line which had acquired resistance as a consequence of repeated 5‐day exposures to stepwise‐increasing concentrations of 5‐FU in vitro. NUGC‐3/5FU/L was 200‐fold and over 16‐fold resistant to 96‐h and 1‐h exposures to 5‐FU, respectively. NUGC‐3/5FU/L incorporated less 5‐FU into RNA, indicating resistance to the RNA‐directed action of 5‐FU. On the other hand, NUGC‐3/5FU/L also showed resistance to in situ thymidylate synthase (TS) inhibition by 5‐FU. Polymerase chain reaction‐single‐strand conformation polymorphism analysis of TS cDNA and a FdUMP ligand binding assay showed that quantitative and qualitative alterations of TS are not responsible for this resistance. In contrast, the ability to metabolize 5‐FU to its active metabolites, FUTP and FdUMP, was reduced in NUGC‐3/5FU/L. We found that not only the activities of uridine phosphorylase/kinase and orotate phosphoribosyl‐transferase (OPRT), but also the level of phosphoribosyl pyrophosphate, a cosubstrate for OPRT, were significantly lower in NUGC‐3/5FU/L than in the parent NUGC‐3. These results indicated that resistance to 5‐FU in NUGC‐3/5FU/L is due to reduced activities of 5‐FU‐anabolizing enzymes, but not to an alteration of TS. 2′‐Deoxyinosine effectively enhanced TS inhibition by 5‐FU in the resistant cells, thus markedly sensitizing them to 5‐FU.
Induction of p53‐independent Apoptosis Associated with G2M Arrest Following DNA Damage in Human Colon Cancer Cell Lines
Authors: Arita Daisaku , Kambe Mariko , Ishioka Chikashi , Kanamaru Ryunosuke
Published: August 2005

The tumor suppressor p53 protein induces apoptosis in response to various kinds of DNA damage in normal cells, but it is still unclear whether or not apoptosis induced by DNA damage correlates with the p53 status in tumor cells. We determined the status of p53 by functional analysis of separated alleles in yeast in five human colon cancer cell lines, SW‐480, SW‐620, DLD‐1, COLO320 and LS174T and investigated whether p53 is necessary for apoptosis and cell cycle arrest after treatment of the cells with a DNA‐damaging agent, etoposide (VP‐16), or γ‐irradiation. Of these cell lines, only LS174T expresses a functional p53. Apoptosis was detected in SW‐480 and COLO320 cell lines, but not in the other cell lines, including LS174T cell line with a normal p53 function. Furthermore, cell cycle analysis revealed accumulation in the G2M phase preceding induction of apoptosis in SW‐480 and COLO320 cells, but not in the other cells. These results suggest that apoptotic induction by DNA damage is not necessarily related to p53 status and that induction of p53‐independent apoptosis following DNA damage may correlate with G2M arrest in the cell cycle, at least in the colon cancer cell lines used in this study.
Helicobacter pylori Extracts Exhibit Nicotinamide Adenine Dinucleotide‐derived Adenylation but Not Mono(adenosine 5‐diphosphate‐ribosyOation of DNA Ligase
Authors: Nozaki Tadashige , Masutani Mitsuko , Noda Takeshi , Saito Daizo , Sugiyama Toshiro , Takato Tsuyoshi , Wakabayashi Keiji , Nakagama Hitoshi , Sugimura Takashi
Published: August 2005

The issue of toxins produced by Helicobacter pylori (H. pylori) urgently requires clarification given that the bacterium causes gastric epithelial cell damage which may lead to precancerous and cancerous changes. During an investigation of the possibility of mono(adenosine 5′‐diphosphate (ADP)‐ribosyl)‐ation by H. pylori products, as observed for other bacterial toxins, we found that radioactivity of [adenylate‐32P]nicotinamide adenine dinucleotide (NAD) is incorporated into an H. pylori protein of 80 kDa after incubation with crude bacterial extract. In contrast, [carbonyl‐14C]NAD did not show any radioactivity incorporation. Unexpectedly, treatment of the modified protein with 0.1 N HC1, but not 0.1 JV NaOH, released the AMP moiety. Such chemical properties are characteristic of bacterial DNA ligase‐AMP complexes. We found that an antibody raised against Escherichia coli DNA ligase [EC 6.5.1.2] immunoprecipitated the modified 80 kDa protein. Our results indicate that incorporation of radioactivity derived from NAD into the 80 kDa protein was due to adenylation, but not mono‐(ADP‐ribosyl) ation, of the DNA ligase of H. pylori.
Hepatitis C Virus in Pelvic Lymph Nodes and Female Reproductive Organs
Authors: Sugiyama Kazuo , Kato Nobuyuki , Ikeda Masanori , Mizutani Tetsuya , Shimotohno Kunitada , Kato Tomoyasu , Sugiyama Yuko , Hasumi Katsuhiko
Published: August 2005

Based on epidemiological evidence, hepatitis C virus (HCV) is thought to be involved in the etiology of not only hepatocellular carcinoma, but also lymphoproliferative diseases. In addition, our previous studies using recently developed cell culture systems that support HCV replication have indicated that HCV possesses both hepatotropism and lympholropism. To determine whether HCV is present in extrahepatic organs, we conducted semi‐quantitative reverse transcription‐polymerase chain reaction for the 5 non‐coding region of the HCV genome in surgical specimens (lymph nodes, ovary, uterus, peripheral blood mononuclear cells [PBMCs] and serum) from three patients with gynecological cancer. We found relatively high HCV genome titers in the lymph nodes, not in the sera, irrespective of various titers in PBMCs. These results suggest that lymph nodes may play an important role in the carrier state and the persistence of HCV infection. Moreover, contrary to expectation, high titers of the HCV genome were observed in the ovaries and the uteri, suggesting the feasibility of mother‐toinfant and spouse‐to‐spouse transmissions of HCV.
P16INK4Gene Mutations Are Relatively Frequent in Ampullary Carcinomas
Authors: Imai Yasuo , Tsurutani Naomi , Oda Hideaki , Nakatsuru Yoko , Inoue Tohru , Ishikawa Takatoshi
Published: August 2005

A high incidence of gene mutations or deletions of P16INK4, a cell cycle regulator which inhibits the activity of cyclin‐dependent kinase 4/cyclin D complex and blocks the Gl‐to‐S transition, has been reported in pancreato‐biliary tract cancers. In order to investigateP16INK4 gene alterations in sporadic ampullary carcinomas, 17 sporadic ampullary carcinomas were examined. After histological diagnosis, DNA samples extracted separately from both cancerous and normal paraffin‐embedded tissues were investigated. Loss of heterozygosity (LOH) was investigated utilizing 3 microsatellite markers on 9p21‐22, and a mutationsl analysis was performed by cloning and sequencing. LOH was observed in 3 cases (17.6%) and somatic mutations with retention of heterozygosity were found in 7 cases (41.2%). Of note was that two mutations resulted in truncated incomplete proteins and one was a point mutation at the consensus site in the conserved ankyrin repeats, which would be crucial for function. Although two‐hit inactivation was not evident in any of the mutation cases and further investigation would be needed to elucidate the role of altered p16INK4, these results suggest that the pl6INK4 gene mutations are relatively frequent and its inactivation might be important in ampnllary carcinogenesis.
Antitumor Activity of a Novel Quinoline Derivative, TAS‐103, with Inhibitory Effects on Topoisomerases I and II
Authors: Utsugi Teruhiro , Aoyagi Kumio , Asao Tetsuji , Okazaki Shinji , Aoyagi Yoshimi , Sano Masaki , Wierzba Konstanty , Yamada Yuji
Published: August 2005

A novel quinoline derivative, TAS‐103 (6‐[[2‐(dimethyIamino)ethyl]amino]‐3‐hydroxy‐7H‐indeno[2,l‐c]quinolin‐7‐one dihydrochloride), was developed as an anticancer agent targeting topoisomerases (topo) I and II, with marked efficacy in solid tumors. TAS‐103 inhibited topo I and II (IC50: 2 μM, 6.5 μM) at a concentration similar to or lower than those of previous agents, and had a strong cytotoxic effect on P388 and KB cells (IC50,: 0.0011 μM, 0.0096 μM). TAS‐103 stabilized topo I and II‐DNA cleavable complexes in KB cells, generating a similar amount of topo II‐DNA complex to that induced by etoposide (VP‐16) but a smaller amount of topo I‐DNA complex than that produced by camptothecin (CPT). In the in vivo study, intermittent i.v. administration was markedly effective against s.c.‐implanted murine tumors. Furthermore, TAS‐103 had marked efficacy against various lung metastatic tumors, and a broad antitumor spectrum in human tumor xenografts (derived from lung, colon, stomach, breast, and pancreatic cancer). The efficacy of TAS‐103 was generally greater than that of irinotecan (CPT‐11), VP‐16, or cis‐diamminedichloroplatinum (CDDP).
Efficient Retrovirus‐mediated Cytokine‐gene Transduction of Primary‐cultured Human Glioma Cells for Tumor Vaccination Therapy
Authors: Wakimoto Hiroaki , Yoshida Yoko , Aoyagi Masaru , Hirakawa Kimiyoshi , Hamada Hirofumi
Published: August 2005

In order to realize a novel vaccination treatment for malignant gliomas using tumor cells genetically modified to express certain cytokines, it is essential to achieve an efficient gene transduction into primary cultured cells. We investigated the feasibility of preparing a glioma vaccine through retro‐virus‐mediated gene transduction. Glioma cells were cultured primarily from surgically resected tumor tissues of six patients. We obtained more than 1000‐fold proliferation of cultures within eight weeks in all six cases. In vitro infection with a recombinant retrovirns GKlacZ carrying an Escherichia coli β‐galsictosidase marker gene resulted in over 65% gene transfer to the primary cultured glioma cells. Further enrichment (∼90%) of transduced cells was possible by employing repeated infections or using vectors with neo marker gene. Two cytokine genes, granulocyte‐macrophage colony‐stimulating factor and interleukin‐4, were introduced into glioma cells by sequential transduction with two single‐expression GK vectors. The transduced glioma cells produced high levels of both cytokines. We also evaluated simultaneous introduction of two genes with double‐expression GK vectors containing internal ribosomal entry site (IRES) or internal promoter (PGK). Although the cells transduced with double‐expression vectors secreted both cytokines, the level of the gene product following IRES or PGK was 10 times lower than that of the upstream gene product. The transduced cells retained cytokine secretion in vitro for 14 days after 100 Gy irradiation. Our data indicate the feasibility of retrovirus‐mediated preparation of gene‐modified tumor vaccines from clinical glioma materials, which could be useful for potentiating antitumor immunity in glioma patients.
Analysis of the DPC4 Gene in Gastric Carcinoma
Authors: Nishizuka Satoshi , Tamura Gen , Maesawa Chihaya , Sakata Ken , Suzuki Yasushi , Iwaya Takeshi , Terashima Masanori , Saito Kazuyoshi , Satodate Ryoichi
Published: August 2005

Allelic loss on chromosome 18q is involved in the progression of gastric carcinoma. Recently, DPC4 (deleted in pancreatic carcinoma, locus 4), a candidate tumor suppressor gene, has been localized at 18q21.1. This gene is inactivated in nearly one half of pancreatic carcinomas. We tested for DPC4 gene mutations and allelic status at 18q21 in 30 primary gastric carcinomas and 5 gastric carcinoma cell lines. Polymerase chain reaction single‐strand conformation polymorphism and sequencing analyses revealed no DPC4 mutations in any of the primary tumors or cell lines. Homozygous deletion ofDPC4 was observed in only 1 (MKN‐45) of the 5 (20%) cell lines. This suggests that the target gene for loss on 18q is not DPC4. The true tumor suppressor gene, encoded near DPC4, has yet to be identified.
Detection of Residual Host Cells in Sex–mismatched Bone Marrow Transplantation in Various Hematological Diseases by Fluorescence in situ Hybridization
Authors: Arif Mansyur , Tanaka Kimio , Kumaravel Tirukalikundram Subrahmanyan , Eguchi Mariko , Iwato Kohji , Dohy Hiroo , Kamada Nanao
Published: August 2005

Thirty–eight sex–mismatched bone marrow transplantation patients with various hematological diseases were followed–up using fluorescence in situ hybridization. Probes specific for various transloca–tions, the X chromosome (DXZ1) and the whole Y chromosome (WCPY), were used to assess successful engraftment and residual host cells. The combination of translocation and WCP Y probes enabled the identification of host and donor cells in addition to the identification of malignant vs. normal cells in the transplant recipient. Fifteen patients were sequentially followed up. The results obtained using the combination of translocation plus WCP Y probes were more reliable than those with DXZ1 plus WCP Y probes, or the translocation probe alone, especially when the percentage of residua] leukemic cells detected by the translocation probe alone was around the cut–off level.
Cell‐killing Efficiency and Number of Platinum Atoms Binding to DNA, RNA and Protein Molecules of HeLa Cells Treated with Combinations of trans‐Diammedichloroplatinum(II) and Hyperthermia
Authors: Akaboshi Mitsuhiko , Kawai Kenichi , Kinashi Yuko , Masunaga Shinichiro , Ono Koji
Published: August 2005

The effect of hyperthermia on the cell‐killing efficiency of Pt atoms binding to DNA, RNA and protein molecules was examined. HeLa S‐3 cells were treated with 195mPt‐radiolabeled trans‐diaminedichloro‐platinum(II) (TDDP) for 60 min at various temperatures, and the relationship between the lethal effect and the number of Pt atoms binding to DNA, RNA and proteins was examined. The mean lethal concentration (D0) of TDDP for a 60‐min treatment at 0, 25, 37, 40, 42 and 44°C was 1714,1016, 302, 179,125 and 42.5 μM, respectively. (D0 is defined as the dose that would give an average of one lethal event per member of the population; for further details, see Fig. 1). By using identically treated cells, the numbers of Pt‐atoms combined with DNA, RNA and protein molecules were determined in the subcellular fractions. Thus, the D0's given as the drug concentrations were replaced with the number of Pt‐atoms combined in each fraction. Then, the cell‐killing efficiency of the Pt atom was expressed as the reciprocal of the number of Pt‐atoms combined and was calculated for each molecule. The efficiency for DNA was 0.206, 0.273, 0.779,1.28,1.77 and 5.14×103 nucleotides, respectively, for the conditions described above. It seemed that hyperthermia potentially interacted not only with bifunctional, but also with monofunctional bonds. Thus, it was concluded that TDDP was markedly less cytotoxic than cis‐diaminedichloroplatinum (CDDP) at 37°C, but was more cytotoxic than CDDP at 44°C.
Assessment of Preference for Breast Cancer Chemopreyention in Japanese Young Women
Authors: Nagata Chisato
Published: August 2005

Pills containing estrogen and progesterone or gonadotropin releasing hormone agonist have been considered valuable to prevent breast cancer. This study assessed preference for the combination‐type pill for preventing breast cancer, to evaluate the hypothetical preventive effect of this agent among young Japanese women. The standard gamble method was applied. Fifty‐five college students and 44 nursing school students aged between 18 and 41 years were asked to decide the probability of being affected by breast cancer at which they would start to take this agent. Preference score was calculated by subtracting the probability given by each respondent from 1, which corresponds to the value (utility) she allotted to the agent. The means of preference score were 0.58, 0.48, 0.37, and 0.27 for 100, 75, 50, and 25% of efficacy levels of the agent, respectively. Preference score was significantly lower in nursing school students and those whose knowledge about hormones were relatively high. Score of Health Locus of Control (HLC) was nonsignificantly negatively correlated with preference score at any efficacy level. HLC score was significantly higher among those who refused the agent with 50 and 25% efficacy levels at 100% level of breast cancer risk. The data suggest that perceived risk of this agent was not negligibly small in this population and school status, knowledge about hormones, and beliefs about health would affect preference for the agent. Understanding of preference for chemopreventive agents for breast cancer, especially those containing hormones, is important to assess their potential as future preventive agents and is helpful when planning a strategy of chemoprevention.
Coexpression of Multiple Sertoli Cell and Leydig Cell Marker Genes in the Spontaneous Testicular Tumor of F344 Rat: Evidence for Phenotypical Bifurcation of the Interstitial Cell Tumor
Authors: Kondoh Gen , Yomogida Kentaro , Dohmae Kayoko , Nozawa Masahiro , Koga Minoru , Nonomura Norio , Miki Tsuneharu , Okuyama Akihiko , Nishimune Yoshitake
Published: August 2005

The development of testicular tumor has been frequently observed in some laboratory rat strains. In the present study, we have further characterized the testicular tumor that spontaneously develops in the F344 rat (F344/Jcl). Tumor cells first appeared in the interstitium and developed into multifocal nodular lesions. In the later stage, the whole testes were occupied by tumor cells that consisted of three different types of cells in morphological appearance: large clear type, small eosinophilic type and intermediate type. To determine the character of these cells, we examined the expression of marker genes for Sertoli cells (e.g., transferrin) and Leydig cells (e.g., 3β–hydroxysteroid dehydrogenase 1 (3β–HSD 1)). Transferrin and 3β–HSD 1 mRNAs were found in all 8 tumor samples analyzed hy northern blotting. By in situ hybridization, we observed a substantial amount of 3β–HSD 1 mRNA and little or no transferrin mRNA in the large clear cells. In contrast, the small eosinophilic cells showed little or no 3β–HSD 1 mRNA and a large amount of transferrin mRNA, suggesting that the tumor was a mixture of at least two types of cells. Other Sertoli cell marker genes, such as cyclic protein 2 and sulfated glycoprotein 2, were expressed in all 8 tumors analyzed, and testin and steel factor (SLF), the c–kit receptor ligand, were also expressed in some of the tumors (testin, 75%; SLF, 25%), while other Leydig cell markers, LH receptor and c–kit, were expressed in 87% and 80% of the tumors, respectively. These results indicate that the spontaneous testicular tumor of F344 rat is of interstitium origin, showing phenotypical bifurcation possibly via transdifferentiation.
Frequent Somatic Mutations of the APC and p53 Genes in Sporadic Ampullary Carcinomas
Authors: Imai Yasuo , Oda Hideaki , Tsurutani Naomi , Nakatsuru Yoko , Inoue Tohru , Ishikawa Takatoshi
Published: August 2005

Although a close relation of somatic mutations of the adenomatous polyposis coli gene with ampullary carcinomas in familial adenomatous polyposis patients has been reported, the possible association with sporadic ampullary neoplasms has not been fully examined. We have therefore investigated loss of heterozygosity at the adenomatous polyposis coli locus and the mutational status of a portion of the adenomatous polyposis coli gene, including the mutation cluster region, in 17 ampullary carcinomas of non–familial adenomatous polyposis patients. Alteration of the adenomatous polyposis coli gene was found in 8 of 17 (47.1%) cases, as missense or insertion mutations, with or without loss of heterozygosity. Additional investigation of ⁁53 (cxons 5–8) and K–ras (codons 12 and 13) gene mutations revealed a striking mutational pattern of thep53 gene. Nine of the 17 cases demonstrated a total of 12 mutations, 6 clustered at codon 189 and 3 at codon 166. Furthermore, 5 of the 12 mutations were nonsense mutations. Regarding the K–ras gene, 4 of the 17 (23.5%) cases had mutations in codon 12, 3 of the 4 cases being derived from the intraduodenal bile duct. The findings indicate that alterations of the adenomatous polyposis coli and the p53 genes are relatively frequent in sporadic ampullary carcinomas. In particular, the clustering at specific p53 codons might offer an etiological clue to clarify ampullary carcinogenesis. Mutations of the K–ras gene, on the other hand, might be characteristic of intraduodenal bile duct origin.
General practitioners’ and family physicians’ negative beliefs and attitudes towards discussing smoking cessation with patients: a systematic review
Authors: Vogt Florian , Hall Sue , Marteau Theresa M.
Published: August 2005

Objective  To estimate the proportion of general practitioners (GPs) and family physicians (FPs) with negative beliefs and attitudes towards discussing smoking cessation with patients.

Methods  A systematic review.

Study selection  All studies published in English, in peer‐reviewed journals, which allowed the extraction of the proportion of GPs and FPs with negative beliefs and attitudes towards discussing smoking cessation.

Data synthesis  Negative beliefs and attitudes were extracted and categorised. Proportions were synthesized giving greater weight to those obtained from studies with larger samples. Those assessed in two or more studies are reported.

Results  Across 19 studies, eight negative beliefs and attitudes were identified. While the majority of GPs and FPs do not have negative beliefs and attitudes towards discussing smoking with their patients, a sizeable minority do. The most common negative beliefs were that such discussions were too time‐consuming (weighted proportion: 42%) and were ineffective (38%). Just over a quarter (22%) of physicians reported lacking confidence in their ability to discuss smoking with their patients, 18% felt such discussions were unpleasant, 16% lacked confidence in their knowledge, and relatively few considered discussing smoking outside of their professional duty (5%), or that this intruded upon patients’ privacy (5%), or that such discussion were inappropriate (3%).

Conclusions  In addition to providing skills training, interventions designed to increase the implementation of smoking cessation interventions by primary care physicians may be more effective if they address a range of commonly held negative beliefs and attitudes towards discussing smoking cessation. These include beliefs and values that influence primary care physicians’ judgements about whether discussing smoking is an effective use of their time.
Allergenic reactivity of the melon profilin Cuc m 2 and its identification as major allergen
Authors: López‐Torrejón G. , Crespo J. F. , Sánchez‐Monge R. , Sánchez‐Jiménez M. , Alvarez J. , Rodriguez J. , Salcedo G.
Published: August 2005

Background: Melon allergy is commonly associated with oral allergy syndrome (OAS) and with hypersensitivity to pollens and other plant foods. No melon allergen responsible for these clinical characteristics has yet been isolated, although profilin has been proposed as a potential target.

Objective: To isolate natural and recombinant melon profilin, to evaluate its in vivo and in vitro reactivity, and to analyse its behaviour in simulated gastric fluid (SGF) and heat treatments.

Methods: A pool or individual sera from 23 patients, and an additional group of 10 patients, all of them with melon allergy, were analysed by in vitro and in vivo tests, respectively. Natural melon profilin (nCuc m 2) and its recombinant counterpart (rCuc m 2) were isolated by poly‐l‐proline affinity chromatography, and characterized by N‐terminal amino acid sequencing, matrix‐assisted laser desorption/ionization analysis, DNA sequencing of cDNAs encoding rCuc m 2, and immunodetection with anti‐profilin antibodies. In vitro analysis included IgE immunodetection, specific IgE determination, ELISA‐inhibition assays, and histamine release (HR) tests. In vivo activity of nCuc m 2 was established by skin prick testing (SPT). The effect of SGF and heat treatment on rCuc m 2 was followed by immunodetection, ELISA inhibition, and HR assays.

Results: Both purified forms of melon profilin were recognized by rabbit anti‐profilin antibodies and IgE of sera from allergic patients, and showed molecular sizes typical of the profilin family. nCuc m 2 had a blocked N‐terminus, whereas rCuc m 2 rendered the expected N‐terminal amino acid sequence, its full protein sequence being highly similar (98–71% identity) to those of profilins from plant foods and pollens. The natural allergen displayed a slightly higher IgE‐binding capacity than its recombinant counterpart. Specific IgE to nCuc m 2 and rCuc m 2 was found in 100% and 78% of the 23 individual sera analysed, respectively. nCuc m 2 evoked positive SPT responses in all (10/10) patients tested, and rCuc m 2 induced HR in two out of three sera assayed. SGF treatment readily inactivated rCuc m 2, as shown by its loss of recognition by anti‐profilin antibodies, lack of IgE binding, and inability to induce HR. In contrast, heat treatment did not affect the IgE‐binding capacity of rCuc m 2.

Conclusions: Profilin is highly prevalent in melon‐allergic patients, and promptly inactivated by SGF, as expected for an allergen mainly linked to OAS.
Predictors of health‐related quality of life in hypertensive recipients following renal transplantation
Authors: Aasebø Willy , Midtvedt Karsten , Hartmann Anders , Stavem Knut
Published: August 2005

Abstract:  Background:  Health‐related quality of life (HRQoL) improves after renal transplantation. However, it is unclear which variables are the strongest determinants of HRQoL following renal transplantation. In this study, we wanted to assess whether antihypertensive medication, donor type, human leukocyte antigen (HLA)‐compatibility or other variables could predict HRQoL 6–12 months after transplantation.

Methods:  The study was a follow up of 124 patients recruited to a single center, randomized, double‐blind clinical trial, comparing the effects of lisinopril and nifedipine in hypertensive renal transplant recipients. HRQoL was assessed with the Short Form 36 (SF‐36) questionnaire. Bivariate and multiple linear regression analysis were used to assess the relationship between potential predictors and the physical component summary (PCS) and mental component summary (MCS) scales of the SF‐36.

Results:  Average scores 6–12 months after transplantation did not differ between patients randomized to lisinopril or nifedipine, or between cadaveric and living donor recipients on any of the eight SF‐36 scales, or the two summary scales. In multivariate analyses, recipient age (p = 0.01) and cold ischemia time >14.5 h (p = 0.04) were independent predictors of the PCS score. Recipient age (p = 0.05), 2–4 HLA‐AB mismatches (p = 0.05) and donor age (p = 0.03) were independent predictors of the MCS score.

Conclusions:  There was no evidence of differences in HRQoL according to lisinopril or nifedipine, or living vs. cadaveric donor transplantation. HRQoL was significantly reduced with longer cold ischemia time and more than one HLA‐AB mismatches, after adjusting for age. These donor kidneys related issues need confirmation.
Behaviour Modelling: Its Record and Potential in Training and Development
Authors: Robertson Ivan T.
Published: August 2005

Behaviour modelling training is an approach to training systems design which has a strong conceptual base in social learning theory (Bandura, 1977). In the mid 1970s, Goldstein and Sorcher produced a now well‐known book which described the application of behaviour‐modelling techniques to supervisory training. In the last 15 years or so the conceptual basis of behaviour modelling has progressed (Bandura, 1986) and there has been a steady stream of studies (see Decker and Nathan, 1985; Goldstein and Gessner, 1988) investigating the use and potential of the approach in organizational settings. The first half of this article provides an overview of the conceptual basis for behaviour modelling training and examines the extent to which behaviour modelling practice provides a valid operationalization of underlying theory. The remainder of the article is concerned with issues of evaluation and applicability. Research on the use of behaviour‐modelling techniques has shown consistently good reaction and learning data (i.e. trainees like the material and are able to learn from it). Evidence concerning the transfer of learning back to the job (i.e. behaviour change and organization impact) is much more equivocal. The available evidence is reviewed and used to produce an account of the record and potential of behaviour modelling training.