bluuebunny/crossref_metadata_2025 huggingface dataset:
Thermostability of Human Immunodeficiency Virus (HIV‐1) in a Liquid Matrix Is Far Higher than That of an Ecotropic Murine Leukemia Virus
Authors: Yoshikura Hiroshi
Published: August 2005

For prevention of HIV infection, which is fatal to man and has no known remedy, sterilization of contaminated materials is particularly important. Before applying any sterilization procedures, they have to be checked by accurately following the kinetics of plaque reduction. Though this is almost self‐evident, such studies have been few. Here, a microplaque assay of HIV is established using HPB‐ALL human T‐cells immobilized on a poly‐L‐lysine‐coated plastic dish. This assay was used to compare the ultraviolet and heat inactivation kinetics of HIV (titrated by this method) with those of Moloney murine leukemia virus (MLV) in a liquid matrix. Though the ultraviolet sensitivities of these viruses were identical (D10=2,800 ergs/mm2), HIV was far more resistant to high temperatures (50°C 70°C) than MLV. This implies that these two viruses have different virion structures, though both are members of retroviridae. The higher thermostability of HIV should be taken into account when HIV‐contaminated materials are handled.
Detection of Human Papillomavirus Type 16 DNA and Papillomavirus Genus‐specific Antigens in Vulva and Cervix from Patients with Bowenoid Papulosis
Authors: Sugase Motoyasu , Moriyama Shin‐ichi , Hata Satoru , Matsukura Toshihiko
Published: August 2005

The warty disordered lesions of the vulva in three female patients were diagnosed as Bowenoid papulosis on the basis of clinical and histopathological findings. In all three vulvar lesions, human Papillomavirus type 16 (HPV 16) DNA was identified hy Southern blot hybridization and Papillomavirus genus‐specific (PGS) antigen was detected in one case immunohistochemically. Furthermore, colposcopic examination revealed the presence of abnormal uterine cervical lesions in two cases. They were found to be intraepithelial neoplasia which harbored HPV 16 DNA and were positive for PGS antigen.
Synthesis of an Inhibitor of Human Immunodeficiency Virus Infection
Authors: Hatanaka Kenichi , Yoshida Takashi , Uryu Toshiyuki , Yoshida Osamu , Nakashima Hideki , Yamamoto Naoki , Mimura Toru , Kaneko Yutaro
Published: August 2005

Anti‐HIV effects of lentinan sulfate were investigated by using an HTLV‐I‐carrying cell line, MT–4, in vitro. Lentinan, a fungal branched (1→3)‐β‐d‐glucan, was sulfated to various degrees by means of two kinds of procedures using piperidine N‐sulfonic acid in dimetbyl sulfoxide or chlorosulfonic acid in pyridine. Lentinan sulfate with a sulfur content of more than 13.9% effectively prevented HIV‐induced cytopathic effects (CPE) at concentrations of more than 3.3 μg/ml. However, low‐substituted lentinan sulfate did not prevent HIV‐induced CPE at any concentration tested. When the counter‐cation was 50% Na± and 50% pyridinium ion, the inhibitory capacity was low. Anticoagulant activity of the lentinan sulfate was also assessed.
Potentiation of Invasive Capacity of Rat Ascites Hepatoma Cells by Transforming Growth Factor‐β
Authors: Mukai Mutsuko , Shinkai Kiyoko , Komatsu Keiko , Akedo Hitoshi
Published: August 2005

The in vitro invasive capacity of poorly invasive cells (W1), which were cloned from rat ascites hepatoma cells (AH 130), was potentiated dose‐ and time‐dependently by pretreating the cells with transforming growth factor‐β (TGF‐β). This potentiation of invasive capacity was completely abolished by anti‐TGF‐β antibody. When the treated cells were ip inoculated into rats, the cells extensively invaded the peritoneum, and formed penetrating tumor nodules. The effect of TGF‐β was reversed by subculturing the treated cells without TGF‐β. The potentiation of invasive capacity by TGF‐β might participate in platelet‐associated enhancement of tumor cell metastasis.
Construction of Mouse A9 Clones Containing a Single Human Chromosome (X/Autosome Translocation) via Micro‐cell Fusion
Authors: Koi Minoru , Morita Hiroyuki , Shimizu Motoyuki , Oshimura Mitsuo
Published: August 2005

Cell hybrids between hypoxanthine guanine phosphoribosyl transferase (HGPRT)‐deficient mouse cell lines (A9 or RAG) and each of 12 different human flbroblasts (GM cells) containing various X/autosome translocations were formed, selected and isolated. Several human chromosomes including an X/autosome translocation carrying HGPRT locus were found in these hybrid cells. To construct A9 cell clones that contain a single X/autosome translocation, micro‐cell fusion was undertaken to transfer these chromosomes from the hybrids to A9 cells. Karyotype analysis revealed that most of the resulting micro‐cell hybrids contain, in a background of mouse chromosomes, only the human X/autosome translocations which were present in the GM cells used for cell hybridization. Sublines of A9 cells were established containing the following autosomal segments: 1q23→1qter; 1q12→1pter; 3p12→3pter; 3q21→3qter; 11q13→11qter; 11q13→11pter; 11p11→11qter; 11q23→11pter; 12q24→12pter; 16q24→16pter; 17q11→17pter.
Stimulation of the Growth of Metastatic Clones of Mouse Colon Adenocarcinoma 26 in vitro by Platelet‐derived Growth Factor
Authors: Tsuruo Takashi , Watanabe Masahiko , Oh‐hara Tomoko
Published: August 2005

The induction of platelet aggregation by tumor cells was found to be an important determinant for the formation of metastasis of a highly metastatic variant of mouse colon adenocarcinoma 26. We found that the growth of highly metastatic clones, NL‐17 and NL‐33, of mouse colon 26 was well stimulated by platelet‐derived growth factor (PDGF) and the stimulation was dependent on the concentration of the growth factor. The growth of weakly metastatic clones, NL‐4 and NL‐44, was stimulated marginally by PDGF, Other factors such as transforming growth factor β and epidermal growth factor did not stimulate the growth of metastatic clones. As the amounts of the receptor of PDGF, as determined by [125I]PDGF binding and mRNA expression, were almost equal in NL‐17 and NL‐44 clones, the difference in growth potential of these clones after the treatment with PDGF could be explained by post‐receptor mechanism(s). The present findings indicate that when tumor cells are arrested in a capillary through the formation of aggregates with platelets, PDGF might play an important role in the establishment of metastasis of mouse colon 26.
Antitumor Activity of BOF‐A2, a New 5‐Fluorouracil Derivative
Authors: Fujii Setsuro , Fukushima Masakazu , Shimamoto Yuji , Ohshimo Hideyuki , Imaoka Takeshi , Shirasaka Tetsuhiko
Published: August 2005

A compound containing both CNDP (3‐cyano‐2,6‐dihydroxypyridine), an inhibitor of 5‐fluorouracil (5‐FU) degradation, and EM‐FU (1‐ethoxymethyl‐5‐fluorouracil), a masked form of 5‐FU, was synthesized and named BOF‐A2 (3‐[3‐(6‐benzoyloxy‐3‐cyano‐2‐pyridyloxycarbonyl)benzoyl]‐1‐ethoxymethyl‐5‐fluorouracil). The antitumor activity of BOF‐A2 was investigated in sarcoma‐180‐bearing mice and Yoshida sarcoma‐bearing rats. The ED50 (the dose for 50% inhibition) values of BOF‐A2 were 25 mg/kg against sarcoma‐180 and 15 mg/kg against Yoshida sarcoma. In vitro studies showed that BOF‐A2 was rapidly degraded to EM‐FU and CNDP in homogenates of the liver and small intestine of mice and rats, and in sera of mice, rats and human, and the conversion of EM‐FU to 5‐FU occurred only in the microsomal fraction of rat liver in the presence of NADPH. After oral administration of BOF‐A2 at 15 mg/kg to Yoshida sarcoma‐bearing rats, BOF‐A2 was hydrolyzed to EM‐FU, CNDP and 5‐FU, and their maximum concentrations in the blood were 2000 ng/ml, 300 ng/ml and 40 ng/ml, respectively. Moreover when BOF‐A2 was given at the same dose to tumor‐bearing mice and rats, the 5‐FU levels in the tumor tissue increased much more than those in the blood and persisted for more than 8 h, whereas those in the blood decreased more rapidly. This accumulation and maintenance of a high level of 5‐FU in the tumor tissue are concluded to be related to the high antitumor activity of BOF‐A2.
Multi‐step Carcinogenesis Model for Adult T‐cell Leukemia
Authors: Okarmoto Takashi , Ohno Yuko , Tsugane Shoichiro , Watanabe Shaw , Shimoyama Masanori , Tajima Kazuo , Miwa Masanao , Shimotohno Kunitada
Published: August 2005

The age‐specific occurrence of adult T‐cell leukemia (ATL) was analyzed using 357 cases collected during nationwide surveys carried out between 1982 and 1985 in Japan. A simple Weibull distribution function fitted well as a model. The mode of ATL onset was log‐linear in this model and the curves for males and females overlapped completely. The presence of age‐dependent accumulation of leukemogenic events within human T‐cell leukemia virus type 1‐immortalized T cells was suggested prior to the development of ATL, and the approximate number of independent leukemogenic events in ATL is estimated to be five. This stochastic analysis supported a multi‐step carcinogenesis as an appropriate model for ATL.
Hematoporphyrin as a Sensitizer of Cell‐damaging Effect of Ultrasound
Authors: Yumita Nagahiko , Nishigaki Ryuichiro , Umemura Koshiro , Umemura Shin‐ichiro
Published: August 2005

Mouse sarcoma 180 or rat ascites hepatoma (AH) 130 cells were exposed to ultrasound (US; 1.27, 2.21 and 3.18 W/cm2; 1.92 MHz) for up to 60 s in vitro in the presence or absence of hematoporphyrin (Hp; 10, 25 and 50 μg/ml). The cell‐damaging effects of treatments were determined by means of the Trypan Blue dye exclusion test. Hp alone did not show any cell‐damaging effect, whereas US alone damaged 30 and 50% of sarcoma and AH 130 cells, respectively, at the maximum intensity for 60 s. In the presence of 50 μg/ml Hp, US damaged 99 and 95% of the above tumor cells, respectively. These results show that Hp increased the sensitivity of tumor cells to US.
Mucosal Dysplasia of Gallbladder: Isolated and Adjacent Lesions to Carcinoma
Authors: Yamagiwa Hiroshi
Published: August 2005

Isolated dysplasia was found in 46 (54 lesions) of 2,027 gallbladders, which were histologically studied by serial step sectioning. Average age of patients was 63.8 with a male‐to‐female ratio of 1 to 3. These lesions, excluding polypoid lesions of adenoma, were located most frequently in the fundus and body, showing flat granular or irregular appearance. As to the size, 68.6% were smaller than 1 cm in diameter (89% were smaller than 2 cm). Most mild dysplasias were found in small lesions less than 0,5 cm in diameter and moderate ones in lesions of less than 1.5 cm, though severe dysplasias were found in various sizes. Gallstones were found in 86.9% of the cases with dysplasia, and intestinal metaplasia and pseudopyloric gland metaplasia were found in 80.4% and 100%. Dysplasia adjacent to carcinoma was found in 46 of 110 cases (41.8%), and this change was frequently found in lesions at the early stage and well‐differentiated carcinoma. It seems that mucosa) damage caused hy a stone and subsequent inflammation may be important in the histogenesis of intestinal metaplasia and dysplasia.
Human Monoclonal Antibodies against Cytokeratin 18 Generated from Patients with Gastric Cancer
Authors: Abe Tsutomu , Fukumoto Masayuki , Tsuchiya Keiko , Kuramochi Kentaro , Furuta Tadaaki , Togoh Shinji , Nishiyama Kiyoshi , Tsuchiya Shuji
Published: August 2005

By co‐culturing regional lymph node B‐cells and HAT‐sensitive mutant cells obtained from RPMI‐1788 cells, no less than 20,000 Epstein‐Barr (EB)‐transformed colonies were obtained from 32 patients with gastric cancer. From B‐cell cultures generating antibodies reactive with gastric cancer tissues as well as cultured gastric cancer cells, two EB‐transformed cell clones termed C418–59 and C1218–39 were isolated. Both of them produced human IgM‐class antibodies, termed Mab418–59 and Mab 1218–39, respectively. Both antibodies reacted with an antigen with a molecular weight of 45 kd existing in gastric cancer MKN‐45, MKN‐1, and Kato‐III cells, and also with all of 4 adenocarcinomas of the stomach in paraffin sections. The antigen recognized by both antibodies was identified as a kind of cytoskeletal protein, cytokeratin 18, In this study, it was confirmed that B‐cell clones generating autoantibodies against cytokeratin 18 were present in some patients with gastric cancer.
Coamplification of the L‐myc and N‐myc Oncogenes in a Neuroblastoma Cell Line
Authors: Jinbo Takeshi , Iwamura Yukio , Kaneko Michio , Sawaguchi Shigenori
Published: August 2005

The L‐myc, N‐myc and c‐myc genes are members of the myc oncogene family. In particular, L‐myc is novel, and amplification of L‐tnyc is still unknown except in small cell lung carcinoma. We examined L‐myc amplification in 30 human neuroblastomas using Southern blot hybridization, and found that the L‐myc gene was amplified approximately 5‐fold in GOTO, a human neuroblastoma cell line. The N‐myc gene was also amplified approximately 60‐fold and furthermore, over‐expression of L‐myc and N‐myc genes was observed in this cell line. In this report, we describe the coampliflcation of the myc gene family in the GOTO neuroblastoma cell line.
Effect of Freeze‐Thawing Breast Milk on Vertical HTLV‐I Transmission from Seropositive Mothers to Children
Authors: Ando Yoshiya , Kakimoto Kazuhiro , Tanigawa Takuo , Furuki Kazuo , Saito Kensuke , Nakano Shiro , Hashimoto Hiratsugu , Moriyama Ikuko , Ichijo Motohiko , Toyama Takenori
Published: August 2005

Breast feeding is known to be a major cause of vertical transmission of HTLV‐I from mothers to her children. The infectiousness of HTLV‐I in breast milk was reported to be lost during freezing and thawing processes. We therefore administered frozen‐and‐thawed breast milk of HTLV‐I carriers to their babies. Among the 13 babies given the frozen‐and‐thawed breast milk (now 12 months of age), no infection has been found yet. This result suggests that freezing and thawing of breast milk is a promising method for the prevention of vertical HTLV‐I infection to breast‐fed babies.
Various Sodium Salts, Potassium Salts, a Calcium Salt and an Ammonium Salt Induced Ornithine Decarboxylase and Stimulated DNA Synthesis in Rat Stomach Mucosa
Authors: Furihata Chie , Yamakoshi Akira , Takezawa Rumiko , Matsushima Taijiro
Published: August 2005

Studies were made on the possible tumor‐promoting activities of various salts of food additives in the glandular stomach mucosa of F344 male rats after their administration by gastric intubation. Up to 100‐fold increases in ornithine decarboxylase (ODC) activity in the pyloric mucosa of the stomach with maxima after 8 h were observed after administration of sodium acetate at doses of 3.68–13.6 mmol/kg body weight, sodium L‐ascorbate at doses of 8.55–17.1 mmol/kg body weight, Na2CO3 at doses of 4.73–14.2 mmol/kg body weight, sodium L‐glutamate at doses of 12.8–17.1 mmol/kg body weight, sodium sorbate at doses of 8.92–17.1 nmol/kg body weight and (NH4)2SO4 at doses of 7.56–20.1 mmol/kg body weight. Increases of up to 100‐fold in ODC activity with maxima after 16 h were also observed after intubation of KC1 at doses of 10.1–22.0 mmol/kg body weight, K2SO3 at doses of 2.84–8.45 mmol/kg body weight, K2S2O5 at doses of 2.25–6.75 mmol/kg body weight and CaCl2 at doses of 2.0–4.08 mmol/kg body weight. Sodium acetate at a dose of 11.0 mmol/kg body weight, KCl at a dose of 20.1 mmol/kg body weight, K2S2O5 at a dose of 5.40 mmol/kg body weight and CaCl2at a dose of 3.4 mmol/kg body weight induced up to 10‐fold increase in DNA synthesis in the pyloric mucosa of the stomach with maxima after 16–24 h. These results suggest that these salts of food additives may, like NaCl, have tumor‐promoting activities in the pyloric mucosa of rat stomach.
Early Gastric Cancer Induced by N‐Ethyl‐N′‐nitro‐N‐nitrosoguanidine in a Cynomolgus Monkey Six Years after Initial Diagnosis of the Lesion
Authors: Szentirmay Zoltan , Ohgaki Hiroko , Maruyama Keiichi , Esumi Hiroyasu , Takayama Shozo , Sugimura Takashi
Published: August 2005

A signet ring cell carcinoma in the gastric antrum of a Cynomolgus monkey induced by N‐ethyl‐N′‐nitro‐N‐nitrosoguanidine was sequentially studied by endoscopy, biopsy, and autopsy. The carcinoma was first detected on the angulus of the stomach at the 38th month as a slightly elevated lesion. Sixty‐one months later this tumor was found to be still in the “early”(intramucosal) stage. Another, independent, initial gastric cancer was also discovered. This is the first example of an induced gastric carcinoma remaining in the “early’stage during a six‐year follow‐up period after the initial histologic diagnosis.
Simulation of Dynamic Changes of Human T‐Cell Leukemia Virus Type I Carriage Rates
Authors: Oguma Shigeru
Published: August 2005

The human T‐cell leukemia virus type I (HTLV‐I) is transmitted via breast milk, semen, or blood transfusion. The last route was not responsible for HTLV‐I infection before the advent of modern medicine, nor will it be a major route in the future because anti HTLV‐I antibody‐positive blood is now screened out. Thus, the carriage rates in various areas of Japan have to be explained by the former two transmission methods. Based on the relationship between the two modes of transmission and carriage rates, several simulation experiments were performed. These experiments revealed that: (a) No population with a vertical transmission rate lower than 50% can be maintained as endemic for the virus. (b) Slight differences in horizontal transmission rates can cause a large change of the carriage rates. (c) A 1,000‐fold carriage rate difference would become indistinguishable within a hundred generations if both modes of transmission were operating at nearly the same rate. (d) The probability of a formerly non‐endemic population becoming endemic due to a single female carrier is not negligible. (e) Prevention of vertical transmission is much more effective in lessening the carriage rate within a short period of time than is prevention of horizontal transmission. A simulation for a real population is also presented.
Expression of Epidermal Growth Factor, Transforming Growth Factor‐α and Their Receptor Genes in Human Gastric Carcinomas; Implication for Autocrine Growth
Authors: Yoshida Kazuhiro , Kyo Eikai , Tsujino Tetsuhiro , Sano Toshiaki , Niimoto Minoru , Tahara Eiichi
Published: August 2005

The expressions of mRNA for epidermal growth factor (EGF), transforming growth factor‐α (TGF‐α) and EGF receptor (EGFR) genes were examined in 7 human gastric carcinoma cell lines and 15 gastric carcinoma tissues and the corresponding normal mucosas. All of the gastric carcinoma cell lines expressed mRNA for EGFR and TGF‐α genes. TMK‐1 and MKN‐28 cells also expressed EGF mRNA. Production of EGF, TGF‐α and EGFR protein by gastric carcinoma cell lines was also confirmed by EGF and TGF‐α specific monoclonal antibody binding. As for surgical specimens, EGFR and TGF‐α mRNA were detected at high levels in all the tumor tissues. Interestingly, EGF mRNA was detected in 5 (33.3%) of the 15 gastric carcinomas but it was not detected in normal tissues. Moreover, anti‐EGF and anti‐TGF‐α monoclonal antibodies inhibited the spontaneous 3H‐TdR uptake by gastric carcinoma cells. These results suggest that EGF and/or TGF‐α produced by tumor cells act as autocrine growth factors for gastric carcinomas.
Enhancement of Polymorphonuclear Leukocyte‐mediated Tumor Cytotoxicity by Serum Factor(s)
Authors: Araki Akemi , Inoue Tomio , Kimura Seishi , Fukase Shigeru , Sendo Fujiro
Published: August 2005

It has been reported that β‐1,3‐d‐glucan isolated from Alcaligenes faecalis (TAK) promoted tumor cytolysis by mouse polymorphonuclear leukocytes (PMN). We investigated the effect of serum on mouse PMN tumor cytolysis induced by TAK and other PMN stimulators. Addition of fetal calf serum (FCS) to the cytolysis assay enhanced tumor cytolysis by PMN in a dose‐dependent manner. Sera obtained from horses, mice, and rats were also effective enhancers of PMN tumor cytolysis. When FCS was added after the assay was under way, the enhancing effect decreased proportionally to the time elapsed. The enhancing activity was detected over a broad range of fractions with a peak at 170 kD by fractionation on a Superose 6 column. The responsible factor(s) in serum was stable after treatment at 60°C, 30 min or after lowering the pH to 2. Mouse PMN stimulated with TAK increased production of hydrogen peroxide in the presence of FCS.
High Yields of Granulosa Cell Tumors/Luteomas in F344 Rat Ovaries after Transplacental Administration of N‐Nitrosobis(2‐oxopropyl)amine
Authors: Maekawa Akihiko , Onodera Hiroshi , Matsushima Yuko , Nagaoka Takaharu , Todate Asahi , Kanno Jun , Shibutani Makoto , Ogasawara Hiroyuki , Yoshida Junichi , Hayashi Yuzo
Published: August 2005

Ovarian tumors were induced at very high incidence in the offspring of F344 rats receiving 3 subcutaneous injections of 10 mg/kg of N‐nitrosobis(2‐oxopropyl)amine on the 14th, 18th and 20th days of gestation. Histologically, all ovarian tumors were of the granulosa cell tumor and/or luteoma type. Many of them consisted of large, polygonal cells with abundant eosinophilic or vacuolated cytoplasm, arranged in sheets or in a pseudo‐palisaded pattern separated by thin fibrovascular stroma, and they exhibited typical luteoma morphological character. The high yields, and the similarities in morphology as well as putative hormonal influence suggest that this experimental system may serve as a good animal model for granulosa cell tumor and/or luteoma development in women.
Production of Both 8‐Hydroxydeoxyguanosine in Liver DNA and γ‐Glutamyltransferase‐positive Hepatocellular Lesions in Rats Given a Choline‐deficient, l‐Amino Acid‐defined Diet
Authors: Nakae Dai , Yoshiji Hitoshi , Maruyama Hiroshi , Kinugasa Tetsuo , Denda Ayumi , Konishi Yoichi
Published: August 2005

The comparative carcinogenic activities of a choline‐deficient, l‐amino acid‐defined diet (CDAADD) and a purified choline‐deficient diet (CDD) for rat liver were studied in terms of both 8‐hydroxy‐deoxyguanosine induction, a marker of DNA damage induced by oxidative stress, and development of γ‐glutamyltransferase(GGT)‐positive putative preneoplastic lesions, including foci and hyperplastic nodules. Twelve weeks after the beginning of treatment, DNA damage could be detected in the liver DNA of rats receiving either CDAADD or CDD, the degree being significantly greater in the former case. Similarly, while GGT‐positive liver lesions were induced by both CDAADD and CDD, the numbers were higher and the areas of lesions were larger in rats receiving CDAADD than in those given CDD. Histologically, hyperplastic nodules were induced in the livers of animals administered CDAADD whereas only foci were seen in the CDD case. The results thus indicate that oxidative stress might be directly involved in rat liver carcinogenesis by CDD and, to a greater degree, with CDAADD.
A Structural Protein Encoded by the 5′ Region of the Hepatitis C Virus Genome Efficiently Detects Viral Infection
Authors: Kato Nobuyuki , Hijikata Makoto , Ootsuyama Yuko , Nakagawa Masanori , Ohkoshi Showgo , Shimotohno Kunitada
Published: August 2005

A structural region of the hepatitis C virus genome was molecularly cloned. A protein expressed in vitro by transcription followed by translation was precipitated immunologically by sera from patients with chronic hepatitis or hepatocellular carcinoma that were positive for antibody against the non‐structural protein, C100, of hepatitis C virus, but not by sera from healthy persons. Thus, this structural protein should be useful for detection of infection with this virus.
Synergistic Effect in Culture of Bleomycin‐group Antibiotics and N‐Solanesyl‐N, N’‐bis(3,4‐dimethoxybenzyl)ethylenediamine, a Synthetic Isoprenoid
Authors: Tomida Akihiro , Suzuki Hideo
Published: August 2005

Like bleomycin and peplomycin, libromycin, a newly developed bleomycin‐group antibiotic, was potentiated 130‐fold against Chinese hamster V79 cells (V79/S) and 47‐fold against its multidrug‐resistant mutant (V79/ADM) by N‐solanesyl‐N, N’‐bis(3,4‐dimethoxybenzyl)ethylenediamine (SDB‐ethylenediamine) at 10 and 3 μg/ml, respectively. But neocarzinostatin, known to cause DNA strand scission as bleomycins do, was potentiated only twofold. This suggests that the high potentiation by SDB‐ethylenediamine is unique to the bleomycin‐group antibiotics. Isobologram analysis revealed that the combined effect of peplomycin and SDB‐ethylenediamine was highly synergistic. SDB‐ethylenediamine did not increase the intracellular accumulation of [3H]peplomycin in V79/S cells. Analyses by an alkaline elution method demonstrated that single strand scission in DNA of intact V79/S cells caused by 1‐h incubation with peplomycin was greatly stimulated by pre‐ and co‐existence of SDB‐ethylenediamine, but DNA strand breaks in isolated nuclei were not affected. Apparently some cytoplasmic constituent(s) is involved in the potentiation mechanism. SDB‐ethylenediamine did not block the DNA repair which occurred after the removal of peplomycin from the medium. Two fragments of SDB‐ethylenediamine, solanesol (polyprenoid moiety) and a diamine component (verapamil‐like moiety), were not synergistic with peplomycin, even when they were mixed together. This indicates that the steric conformation of the intact SDB‐ethylenediamine molecule is important for the activity.
Frequent Loss of Heterozygosity on Chromosomes 16 and 4 in Human Hepatocellular Carcinoma
Authors: Zhang Wandong , Hirohashi Setsuo , Tsuda Hitoshi , Shimosato Yukio , Yokota Jun , Terada Masaaki , Sugimura Takashi
Published: August 2005

By restriction fragment length polymorphism analysis, we examined loss of heterozygosity at 34 loci on 23 chromosomes in 35 surgically resected human hepatocellular carcinomas. Allele losses at the HP locus on chromosome 16q22 and at the MT2P1 locus on chromosome 4p11‐q21 were detected in 57% (8/14) and 50% (8/16) of cases, respectively. Loss of heterozygosity on chromosomes 16q and 4 occurred simultaneously in 4 of 7 informative cases for both loci, and seemed to be important in the development of human hepatocellular carcinoma irrespective of the presence of hepatitis B virus infection. In contrast, the incidence of allele loss was low at the other loci, e.g., chromosome 1p, 3p, 11p, 13q or 17p, where one allele is frequently lost in other cancers.
Lack of Tumorigenicity of Aminopyrine Orally Administered to B6C3F1 Mice
Authors: Inai Kouki , Kobuke Toshihiro , Fujihara Megumu , Yonehara Shuji , Takemoto Tsuyoshi , Tsuya Takafumi , Yamamoto Atsushi , Tachiyama Yoshiro , Izumi Kumiko , Tokuoka Shoji
Published: August 2005

To test the tumorigenic potential of aminopyrine, an antipyretic analgesic, it was administered in drinking water at levels of 0 (control), 0.04 and 0.08% to 50 male and 50 female B6C3F1, mice for 100 weeks, and the mice were subsequently maintained without aminopyrine for a further 4 weeks. The most frequent types of tumor, in both treated and control groups, were hepatocellular tumor in male mice and malignant lymphoma/lymphoid leukemia in female mice. No statistically significant differences were observed in the incidences of these tumors between treated and control groups. The incidences of several other tumors in male and female mice also showed no statistically significant differences between treated and control groups. Therefore, no tumorigenic effect of orally administered aminopyrine in B6C3F1 mice was apparent in the present study.
Production and Characterization of a Human Monoclonal Antibody Recognizing a New Antigen Expressed on Some Lymphoid Cells
Authors: Ikewaki Nobunao , Tamauchi Hidekazu , Yamaura Noboru , Takahashi Hiroomi , Inoko Hidetoshi
Published: August 2005

A cell line secreting a human monoclonal antibody was established by Epstein‐Barr virus transforming B cells derived from an enlarged cervical lymph node excised from a patient bearing a carotid body tumor. The reactivity of the monoclonal antibody, designated as mNISP, was tested on various cells and cell lines. An antigen defined by the mNISP was expressed on some Burkitt's lymphoma cell lines and on a non‐T non‐B acute lymphoblastic leukemia cell line. Furthermore, this antigen was expressed on leukemic cells from 2 of 8 patients with chronic myelocytic leukemia, 2 of 10 patients with acute myeloblastic leukemia, one of 13 patients with acute lymphoblastic leukemia, and two patients with adult T cell leukemia, but it was not expressed on normal T, B and adherent (macrophage) cells. In addition, mNISP reacted with T cells obtained from human T‐cell leukemia virus type I carriers. We found that the antigen defined by mNISP was distinct from any previously reported antigen in terms of its pattern of cellular expression and molecular weight, suggesting that mNISP recognizes a new antigen expressed on some lymphoid cells.
Comparative Antitumor Activity and Intestinal Toxicity of 5′‐Deoxy‐5‐fluorouridine and Its Prodrug Trimethoxybenzoyl‐5′‐deoxy‐5‐fluorocytidine
Authors: Ninomiya Yasuyuki , Miwa Masanori , Eda Hiroyuki , Sahara Hiroshi , Fujimoto Kaori , Ishida Masako , Umeda Isao , Yokose Kazuteru , Ishitsuka Hideo
Published: August 2005

N4‐Trimethoxybenzoyl‐5′‐deoxy‐5‐fluorocytidine (Ro 09–1390), a prodrug of the cytostatic 5′‐deoxy‐5‐fluorouridine (5′‐DFUR), was synthesized with the aim of reducing of the dose‐limiting toxicity of 5′‐DFUR, which is diarrhea. In mice bearing Lewis lung carcinoma, 5′‐DFUR given po produced a substantial amount of 5‐fluorouracil (5‐FU) in the intestinal tract as well as in tumors, where the enzyme pyrimidine nucleoside phosphorylase, essential for conversion of 5′‐DFUR to 5‐FU, is predominantly located. With the oral administration of Ro 09–1390 only a small amount of 5‐FU was formed in the intestine; however, the administration of Ro 09–1390 and 5′‐DFUR at the same dose produced similar amounts of 5‐FU in tumor tissues. These differences in metabolism were reflected in their toxicity and antitumor efficacy. The administration of 5′‐DFUR resulted in damage to the intestinal mucosal membrane and diarrhea in normal mice, whereas Ro 09–1390 was much less toxic to the intestinal tract. As regards antitumor activity, Ro 09–1390 and 5′‐DFUR at equivalent doses inhibited the growth of Lewis lung carcinoma to similar extents. Since Ro 09–1390 was much less toxic to the intestinal tract than 5′‐DFUR, mice bearing Lewis lung carcinoma could be given Ro 09–1390 daily over a longer period and at a higher dose, resulting in a longer survival time.
Heterogeneity of Antigen Molecules Recognized by Anti‐tax1 Monoclonal Antibody Lt‐4 in Cell Lines Bearing Human T Cell Leukemia Virus Type I and Related Retroviruses
Authors: Tanaka Yuetsu , Yoshida Atsushi , Takayama Yoshinaga , Tsujimoto Hajime , Tsujimoto Atsumi , Hayami Masanori , Tozawa Hideki
Published: August 2005

Using a monoclonal antibody, Lt‐4, directed against human T cell leukemia virus type I (HTLV‐I) trans‐activator (tax1) antigen, we examined the expression of tax1 and related antigens in a variety of T cell lines bearing HTLV‐I and related retroviruses, simian T cell leukemia virus type I (STLV‐I) and HTLV‐II, by immunofluorescence and immunoblot assays. Lt‐4 reacted with all HTLV‐I‐bearing cell lines tested and five out of eight simian cell lines bearing STLV‐I, but not with an HTLV‐II‐bearing cell line. Lt‐4 detected 40 kd tax1 antigen molecules in most HTLV‐I‐bearing cell lines except one cell line that expressed 39 kd tax1 antigen. In the STLV‐I‐bearing T cell lines, tax1‐related antigen molecules detected by Lt‐4 were heterogeneous, having molecular weights in the range of 36–41 kd.
A Simple, General Method for Detecting Retroviral RNAs Expressed in Cells
Authors: Harada Fumio , Hirose Yutaka , Tanaka Motohiro , Sasaki Takuma , Kamiya Hiroyuki , Miura Kazunobu
Published: August 2005

A simple, efficient method has been developed for detecting retroviral RNAs expressed in cells. In this method, total RNAs or poly A+ RNAs extracted from various human cells are separated by electrophoresis and hybridized with synthetic oligonucleotides corresponding to the 3′‐terminal 18 nucleotides of various tRNAs. Genomic and subgenomic RNAs of HTLV‐I and HTLV‐II in virus‐infected cells and of xenotropic murine leukemia virus expressed in human lung cancer cells were easily detected with the tRNAPro‐derived oligonucleotide probe. This technique can be used to search for unidentified retroviruses expressed in human cancer cells and tissues.
Detection of Epstein‐Barr Virus DNA in Reed‐Sternberg Cells of Hodgkin's Disease Using the Polymerase Chain Reaction and in situ Hybridization
Authors: Uhara Hisashi , Sato Yuichi , Mukai Kiyoshi , Akao Ichiro , Matsuno Yoshihiro , Furuya Shuichiroh , Hoshikawa Tomohide , Shimosato Yukio , Saida Toshiaki
Published: August 2005

Thirty‐one cases of Hodgkin's disease were examined for the occurrence of Epstein‐Barr virus (EBV) genome by using the polymerase chain reaction (PCR) of DNA in formalin‐fixed paraffin‐embedded tissues and the in situ hybridization technique. The cases were subdivided into 17 cases of nodular sclerosis (NS), nine cases of mixed cellularity (MC), four cases of lymphocyte predominance (LP), and one case of lymphocyte depletion (LD). EBV DNA was detected in eight cases including four cases of NS, three cases of MC and one case of LP. The sensitivity of PCR was higher than that of Southern blot hybridization of DNA from fresh frozen tissue, because Southern blot hybridization using the BamHI‐W fragment of EBV detected virus DNA only in two of three cases which were positive by PCR. The results of in situ hybridization studies confirmed that EBV genome was localized within the nuclei of Reed‐Sternberg (RS) cells and their mononuclear variants. Furthermore, double‐labeling studies combining in situ hybridization and immunocytochemistry using CD30 (BerH2) and CD15 (LeuM1) as markers of RS cells, as well as pan B‐marker (L26) and pan T‐marker, CD45RO (UCHL1), were performed to demonstrate the phenotype of EBV DNA‐positive cells, confirming that EBV DNA was present in RS cells but not in lymphocytes. The results of this study indicate a significant association between EBV and some cases of Hodgkin's disease.
Expression of ret Proto‐oncogene in Human Neuroblastomas
Authors: Nagao Minako , Ishizaka Yukihito , Nakagawara Akira , Kohno Kimitoshi , Kuwano Michihiko , Tahira Tomoko , Itoh Fumio , Ikeda Isuzu , Sugimura Takashi
Published: August 2005

We examined the expression of ret proto‐oncogene (proto‐ret) in surgically resected human neuroblastomas. Slot blot RNA hybridization revealed that all 29 neuroblastomas examined expressed the proto‐ret, the relative intensity of the hybridization ranging from 1 to 48. No correlation was found between the level of expression of proto‐ret and the clinical stage. The level of expression was also not correlated with N‐myc amplification, the patient's age or the histological type of the tumor. Based on the previous finding that proto‐ret expression is very rarely detected in tumor cell lines other than those of neuroblastoma, proto‐ret expression was suggested to be a characteristic of neuroblastomas, and possibly to be involved in the genesis of neuroblastomas.
Effect of Human Macrophage Colony‐stimulating Factor on Granulopoiesis and Survival in Bone‐marrow‐transplanted Mice
Authors: Yanai Nobuya , Yamada Muneo , Motoyoshi Kazuo , Yokota Hajime , Yoshida Katsuo , Saito Minoru , Kawashima Takuji , Nishida Masayuki , Miura Yasusada , Saito Masaki , Takaku Fumimaro
Published: August 2005

Human macrophage colony‐stimulating factor (hM‐CSF) has been isolated from normal human urine and purified to a homogenous protein. The effect of hM‐CSF on granulopoiesis was investigated in BALB/c mice transplanted with a suboptimal number of bone marrow cells. Lethally irradiated (7.8 Gy) mice were transplanted with 1 × 106 syngeneic mouse bone marrow cells and treated with a daily intraperitoneal dose of 64 μg/kg of hM‐CSF for 5 days following the transplant. The hM‐CSF injection resulted in stimulation of the recovery of blood neutrophils as well as an increase in the number of granulocyte‐macrophage progenitor cells (CFU‐GM) in the femur and spleen. The survival of lethally irradiated mice was dependent on the cell number transplanted; most mice transplanted with 2 × 104 cells died within 2 weeks. The recovery of hematopoiesis in mice transplanted with 2 × 104 cells was modestly but significantly stimulated by hM‐CSF administration initiated from 5 days before or 1 day after transplantation for a 5‐day period. Furthermore, the hM‐CSF administrations markedly reduced the mortality in these mice during the early period after the transplantation. Since anaerobic bacteria were frequently detected in arterial blood immediately before the deaths but were not found in the surviving mice, it is speculated that early deaths occurring within 2 weeks after the transplant may be caused by opportunistic infections, and hM‐CSF injection may prevent these mortal infections through its stimulating effect on monocyte‐macrophage functions that are responsible for the production of hematopoietic regulators.
Comparative Study of the N‐Linked Oligosaccharides Released from Normal Human Esophageal Epithelium and Esophageal Squamous Carcinoma
Authors: Hiraizuni Sen , Takasaki Seiichi , Nishihira Tetsuro , Mori Shouzo , Kobata Akira
Published: August 2005

N‐Linked sugar chains of normal human esophageal epithelium and esophageal squamous carcinoma were quantitatively released as oligosaccharides from their membrane preparations by hydrazinolysis. After being fractionated by serial lectin column chromatography using concanavalin A‐Sepharose and Datura stramonium agglutinin‐Sepharose, their structures were elucidated by exoglycosidase digestion in combination with methylation analysis. Both normal epithelium and esophageal carcinoma contained bi‐, tri‐ and tetraantennary oligosaccharides as well as high mannose‐type oligosaccharides. Interestingly, carcinoma had about 1.6 times larger amounts of tri‐ and tetraantennary oligosaccharides with the GlcNAcβ1→4Manα1→ and/or the GlcNAcβ1→6Manα1→ linkages than normal epithelium. Tri‐ and tetraantennary oligosaccharides with N‐acetyllactosamine repeating units (the Galβ1→4GlcNAcβ1→3Galβ1→4GlcNAc group) were also increased in carcinoma. These data indicated that the altered glycosylation of proteins previously found in transformed rodent cells also occurs widely in human esophageal carcinoma.
Light and Electron Microscopic Study of Vacuolated Cells in Immunoblastic Lymphadenopathy‐like T‐Cell Lymphoma
Authors: Hirose Yuko , Shimizu Shiro , Yoshioka Ritsuko , Tachibana Junko , Sugai Susumu , Takiguchi Tomoo , Konda Susumu
Published: August 2005

Three cases of immunoblastic lymphadenopathy (IBL)‐like T‐cell lymphoma were analyzed immunologically, and the ultrastructure of mononuclear cells in the lymph nodes and peripheral blood was examined. In the peripheral blood, light microscopic examination revealed vacuolated lymphocytes. These vacuolated lymphocytes formed rosettes with sheep erythrocytes, and they were CD3‐ and CD4‐positive using the avidin‐biotin method in cases 1 and 2. Electron microscopic examination revealed two kinds of abnormal lymphocytes. One kind was of B‐cell nature with rich lamellated rough‐surfaced endoplasmic reticulum and mitochondria. The other kind had a large cytoplasm, in which Golgi apparatus, some endoplasmic reticulum, some mitochondria and a few vacuoles were seen. Some of these vacuoles had remnants of mitochondrial cristae or were enlarged endoplasmic reticulum. The vacuolated T lymphocytes and activated lymphocytes of B‐cell nature disappeared with chemotherapy but reappeared with relapse of the disease. These observations suggest that vacuolated cells are related to pale cells in lymph node sections. In other words when these vacuolated cells are found in the peripheral blood of patients with lymphoid malignancies, IBL‐like T‐cell lymphoma can be suspected.
A Domain Responsible for the Transformation Suppressor Activity in Krev‐1 Protein
Authors: Kitayama Hitoshi , Matsuzaki Tomoko , Ikawa Yoji , Noda Makoto
Published: August 2005

Krev‐1 cDNA encodes a ras‐related protein and exhibits an activity of inducing flat revertants at certain frequencies (2‐5% of total transfectants) when introduced into a v‐K‐ras‐transformed mouse NIH3T3 cell line, DT. To explore the functional organization of Krev‐1 protein, we constructed a series of chimeric genes consisting of fragments of H‐ras and Krev‐1 cDNAs, and tested their biological activities in DT cells. The results indicated that the determinant for the transformation suppressor activity resides in the N‐terminal one‐third of the Krev‐1 encoded polypeptide within which a highly conserved, putative effector‐binding domain is present.
Human Platelets Effectively Kill K‐562 Cells, a Chronic Myelogenic Leukemia Cell Line, in vitro
Authors: Sagawa Terutaka , Kodama Takeshi , Tominaga Akio , Okada Mariko
Published: August 2005

The cytotoxic effect of isolated human platelets on leukemic cells has been examined in order to investigate the role of platelets in host defense systems. K‐562 cells (a chronic myelogenic leukemia cell line) showed significant change in their morphology and were killed when they were incubated with platelets in serum‐free medium for several hours at 37°C, a condition where no killing of normal peripheral lymphocytes occurred. Some protease inhibitors inhibited the cytotoxicity of platelets against K‐562 cells. Our results suggest that platelets may be involved in host defense against neoplasia and that certain proteases are implicated in the cytotoxic effect of platelets.
DNA Adducts Formed by 2‐Amino‐3,8‐dimethylimidazo[4,5‐f]quinoxaline in Rat Liver: Dose‐Response on Chronic Administration
Authors: Yamashita Katsumi , Adachi Masaaki , Kato Shunji , Nakagama Hitoshi , Ochiai Masako , Wakabayashi Keiji , Sato Shigeaki , Nagao Minako , Sugimura Takashi
Published: August 2005

The effect of administration of 2‐amino‐3,8‐dimethylimidazo[4,5‐f]quinoxaline (MeIQx) at various doses on DNA adduct formation in male rats was examined by 32P‐postlabeling analysis. Administration of MeIQx in the diet at 0.4 ppm, 4 ppm, 40 ppm and 400 ppm for one week resulted in the formations of 0.04, 0.28, 3.34 and 39.0 adducts per 107 nucleotides in rat liver cells. Continuous administration of 400 ppm of MeIQx in the diet for 61 weeks to rats induced hepatocellular carcinomas in all rats. The carcinogenicity of MeIQx at doses of 40 ppm or less is not known yet, but the above results show a linear relationship between the level of MeIQx administered and the adduct level. In rats treated with low doses of 0.4, 4 and 40 ppm of MeIQx, adduct levels increased linearly with time of treatment, the levels in week 12 being two to three times those in week 1. In contrast, on treatment with 400 ppm of MeIQx, the adduct level in the liver increased until week 4, when it was 110 adducts per 107 nucleotides, and then remained constant for the next 8 weeks. Induction of the multidrug‐resistance gene was suggested to be involved in development of this plateau level.
Urothelial Damage and Tumor Initiation by Urinary Metabolites of Sodium o‐Phenylphenate in the Urinary Bladder of Female Rats
Authors: Hasegawa Ryohei , Furukawa Fumio , Toyoda Kazuhiro , Sato Hidetaka , Takahashi Michihito , Hayashi Yuzo
Published: August 2005

Intravesical instillation of 2‐phenyl‐1,4‐benzoquinone (PBQ), a metabolite of sodium o‐phenylphenate (Na‐OPP), at a concentration of 0.1% in saline caused acute epithelial injury, prolonged inflammation and epithelial hyperplasia in female F344 rats. Ten such instillations of PBQ within 5 weeks resulted in the development of preneoplastic lesions of the urinary bladder epithelium when followed by 5% sodium saccharin feeding for 31 weeks. Since neither urothelial damage nor tumor‐initiating activity was observed with either Na‐OPP itself or another metabolite, phenylhydroquinone, PBQ may play an essential role in Na‐OPP urinary bladder carcinogenesis.
In vitro and in vivo Release of Soluble erbB‐2 Protein from Human Carcinoma Cells
Authors: Mori Shigeo , Mori Yasuyoshi , Mukaiyama Taketo , Yamada Yukinori , Sonobe Yoshiko , Matsushita Hirohisa , Sakamoto Goi , Akiyama Tetsu , Ogawa Makoto , Shiraishi Masato , Toyoshima Kumao , Yamamoto Tadashi
Published: August 2005

An enzyme‐linked immunosorbent assay (ELISA) was developed to measure soluble erbB‐2 protein in culture supernatants of various human cell lines and sera of patients suffering from recurrent breast carcinoma. Soluble erbB‐2 protein was demonstrated in culture supernatants of cell lines that expressed high levels of erbB‐2 protein as shown by western blot analysis of cell lysates. Increased levels of the protein, 40‐ to 190‐fold higher than in healthy controls, were demonstrated in sera of 3 out of 12 patients with breast carcinomas. On immunohistological study of tumor tissues from 9 patients, high immune reaction with the anti‐erbB‐2 protein antibody was observed in 2 cases. These were two of the three patients who had elevated levels of erbB‐2 protein in serum (a sample was not available from the third patient). These results raise the possibility that soluble erbB‐2 protein level in serum can be used as an indicator for spread of carcinomas that overexpress erbB‐2 protein.
Characterization of Carcinoembryonic Antigen‐related Antigens in Normal Adult Feces
Authors: Matsuoka Yuji , Matsuo Yoshino , Sugano Kokichi , Ohkura Hisanao , Kuroki Motomu , Kuroki Masahide
Published: August 2005

About 50–70 mg in total of carcinoembryonic antigen (CEA) (or CEA‐related antigens) was detected in normal adult feces evacuated during one day (200–250 g). Ten percent or less of the antigen was found to be in soluble form in fresh feces (naturally solubilized antigen), while 90% or more was still in membrane‐bound form which was releasable with phosphatidylinositol‐specific phospholipase C (PI‐PLC‐solubilized antigen). The naturally solubilized and PI‐PLC‐solubilized antigens are anti‐genically different from each other and similar to normal fecal antigen (NFA)‐2 and CEA, respectively, suggesting that “CEA‐distinctive’antigenicity detected so far in CEA from cancerous tissues is not due to the difference between antigens in normal and malignant tissues but is probably due to the presence of the glycosylinositolphosphate moiety at the carboxyl‐terminus of the antigen molecule. Thus, “CEA‐distinctive’antigenicity is by no means cancer‐specific, but this antigenicity seems to be critical for the clinical significance of CEA as a tumor marker, because an assay system (Kit II) which is able to distinguish CEA from NFA‐2 revealed much improved features in cancer diagnosis as reported recently.
Prevalence of Antibody against Non‐A, non‐B Hepatitis Virus in Japanese Patients with Hepatocellular Carcinoma
Authors: Ohkoshi Showgo , Kojima Hideo , Tawaraya Hironobu , Miyajima Tohru , Kamimura Tomoteru , Asakura Hitoshi , Satoh Akira , Hirose Shinichi , Hijikata Makoto , Kato Nobuyuki , Shimotohno Kunitada
Published: August 2005

Antibodies against a possible causative agent of non‐A, non‐B hepatitis, hepatitis C virus (HCV), in Japanese patients with hepatocellular carcinoma were analyzed using the enzyme‐linked immunosorbent assay (ELISA) system from Ortho Diagnostic Systems, Japan. Fifty of 58 cases of hepatitis B virus surface antigen (HBsAg)‐negative hepatocellular carcinoma were positive for the antibody (86%) and 8 of 42 cases of HBsAg‐positive hepatocellular carcinoma were positive (19%). Among patients with HBsAg‐negative hepatocellular carcinoma, the prevalence of the antibody was greater among those who had received a blood transfusion (97%) than among those with no history of transfusion (70%). Only 3 of 54 patients with cancers other than hepatocellular carcinoma were found to be antibody‐positive (5.6%) and all three patients had a history of blood transfusion. These results show a close relationship between the presence of anti‐HCV antibody and HBsAg‐negative hepatocellular carcinoma in Japan.