bluuebunny/crossref_metadata_2025 huggingface dataset:
Single‐cell Suspension Assay with an MTT End Point Is Useful for Evaluating the Optimal Adjuvant Chemotherapy for Advanced Gastric Cancer
Authors: Saikawa Yoshiro , Kubota Tetsuro , Furukawa Toshiharu , Suto Akihiko , Watanabe Masahiko , Kumai Koichiro , Ishibiki Kyuya , Kitajima Masaki
Published: August 2005

One hundred and forty‐eight patients with gastric cancer admitted to Keio University Hospital between July 1988 and October 1992 underwent resection of the primary lesion, as well as single‐cell suspension assay of fresh surgical materials with 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT assay) for chemosensitivity evaluation. Fifty patients with histologically stage III or IV gastric cancer were enrolled in this study, among whom 10 received no chemotherapy after surgery while 40 received chemotherapy at equivalent dose levels after surgery. The patients given chemotherapy were divided into two groups consisting of an “Adapted” group treated with at least one agent identified as effective by the assay, and a “Non‐adapted” group treated with agents to which the cells were not sensitive in the assay, in order to identify the optimal cut‐off inhibition rate (IR) in MTT assay for evaluation of the appropriate adjuvant cancer chemotherapy after surgery. A cut‐off IR of 30% was optimal for differentiating the survival rates between the “Adapted” and “Non‐adapted” groups. Patients treated with drugs which showed more than 30% IR on their surgical specimens showed a better survival rate than patients treated with drugs which were ineffective in the assay.
Mitotic Activation of c‐Src Is Suppressed by Csk
Authors: Ohsato Yoshiharu , Nada Shigeyuki , Okada Masato , Nakagawa Hachiro
Published: August 2005

The kinase activity of the proto‐oncogene product, c‐Src, increases during mitosis through partial dephosphorylation of Tyr527, the negative regulatory site of c‐Src. To examine whether or not Csk, a candidate kinasc specific for Tyr527, is involved in this regulation, we developed a BalbJc 3T3 cell line overexpressing Csk and a Csk‐deficient cell line. The overexpression of wild‐type Csk caused significant suppression of the c‐Src activity during mitosis. A membrane‐targeted Csk, which has an ammo‐terminal myristylation signal of c‐Src, exhibited an effective suppression of the c‐Src activity, even though its expression level was lower than that of endogenous Csk. Concomitant with the suppression of the c‐Src activation, the level of tyrosine phosphorylation of a cortactin‐related protein, a potential substrate of c‐Src in vivo, was reduced. In contrast, the Csk‐deficient cells exhibited constitutive activation of c‐Src, which showed no significant change in its activity during mitosis. These results suggest that Csk indeed participates in the regulation of the c‐Src activity during mitosis.
Cancer Mortality in Low Radon Spa Area
Authors: Suzuki Yasuyo , Honjo Satoshi , Kawamura Hiroko , Koishi Fumiko , Suzuki Tomokazu , Hirohata Tomio
Published: August 2005

Recently lower mortality for cancers of all sites was reported among inhabitants in the Misasa spa area, where there is a high radon background. To clarify the effects of radon exposure on cancer mortality, the effects of a hot spring itself on cancer mortality was investigated in the Beppu spa area, which has only a low radon background, and adjacent control areas. For females, the mortalities for cancers of all sites, liver and lung were higher in Beppu than those for all Japan on the basis of the standardized mortality ratio (SMR), while the SMR for all cancers was lower in adjacent areas. For the male inhabitants in hoth areas the cancer mortalities of all sites were not significantly different from those of all Japan. When we directly compared the most typical spa areas in Beppu and an adjacent control area, a Poisson regression analysis did not show that the relative risk of dying from cancer of all sites was decreased in the spa areas. These results are thus consistent with the view that the lower cancer mortality in the Misasa spa area might be related to exposure to low levels of radon

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Enhancing Effects of Organosulfur Compounds from Garlic and Onions on Hepatocarcinogenesis in Rats: Association with Increased Cell Proliferation and Elevated Ornithine Decarboxylase Activity
Authors: Takada Nobuyasu , Kitano Mitsuaki , Chen Tianxin , Yano Yoshihisa , Otani Shuzo , Fukushima Shoji
Published: August 2005

Four organosulfur compounds from garlic and onions were examined for modifying effects on diethylnitrosamine (DEN)‐induced neoplasia of the liver in male F344 rats using the medium‐term bioassay system based on the two‐step model of hepatocarcinogenesis. Carcinogenic potential was scored by comparing the numbers and areas per cm2 of induced glutathlone S‐transfcrasc placental form‐positive foci. Isothiocyanic acid isobutyl ester (IAIE), dipropyl trisulfide (DPT), and allyl mercapton (AM) exerted enhancing effects on their development, while dimethyl trisulfide also tended to increase them. To investigate possible mechanisms of the modifying influence, sequential changes in ornithine decarboxylase activity (ODC) over 24 h were measured in AM‐treated liver tissue without prior DEN initiation. The activity started to increase by 4 h after AM‐treatment, and reached maximum at 16 h, compared to controls. Spermidine/spermine N1‐acetyltransferase activity was not significantly changed. An increase in proliferating cell nuclear antigen‐positive cells followed the elevation of ODC activity. These results suggest that IAIE, DPT, and AM promote rat hepatocarcinogenesis and their promoting effect might be caused by increased cell proliferation with increased poly‐amine biosynthesis. In evaluating relationships between diet and cancer, it is thus appropriate to consider not only a possible protective role of garlic and onions, but also enhancing effects.
A Randomized Cross‐over Study of High‐dose Metoclopramide plus Dexamethasone versus Granisetron plus Dexamethasone in Patients Receiving Chemotherapy with High‐dose Cisplatin
Authors: Ohmatsu Hironobu , Eguchi Kenji , Shinkai Tetsu , Tamura Tomohide , Ohe Yuichiro , Nisio Masato , Kunikane Hiroshi , Arioka Hitoshi , Karato Atsuya , Nakashima Hajime , Sasaki Yasutsuna , Tajima Kinuko , Tada Noriko , Saijo Nagahiro
Published: August 2005

We carried out a randomized, single‐blind, cross‐over trial to compare the antiemetic effect, for both acute and delayed emesis, of granisetron plus dexamethasone (GRN+Dx) with that of high‐dose metoclopramide plus dexamethasone (HDMP + Dx). Fifty‐four patients with primary or metastatic lung cancer, given single‐dose cisplatin (> 80 mg/m2) chemotherapy more than twice, were enrolled in this study. They were treated with both HDMP+Dx and GRN+Dx in two consecutive chemotherapy courses. On day 1, patients experienced a mean of 2.5 (SD=4.3) and 0,1 (SD = 0.4) episodes of vomiting in the HDMP+Dx and the GRN + Dx groups, respectively (P=0.0008). Complete response rate on day 1 was 45 and 90% in the HDMP+Dx and the GRN+Dx groups, respectively (P= 0.0001). Patients treated with GRN+Dx had a tendency to suffer more episodes of vomiting than the HDMP+Dx group on days 2–5, but it was not statistically significant. Twenty‐four patients (57%) preferred the GRN+Dx treatment and 14 patients (33%), HDMP + Dx. In the HDMP + Dx group, nine patients (21%) had an extrapyramidal reaction, and 5 patients (12%) had constipation that lasted for at least two days. In contrast, no patients had extrapyramidal reactions, and IS patients (43%) had constipation in the GRN+Dx group (P < 0.01). GRN+Dx was more effective than HDMP+Dx only in preventing the acute emesis induced by cisplatin. An effective treatment for delayed emesis is still needed.
Risk of Lung Cancer among Cigarette Smokers in Relation to Tumor Location
Authors: Shimizu Hiroyuki , Nagata Chisato , Tsuchiya Eiju , Nakagawa Ken , Weng Siu‐Yuhe
Published: August 2005

To evaluate the effect of cigarette smoking on lung cancer by tumor location and histological type, we compared the smoking history obtained from medical records of 605 patients with squamous cell carcinoma (194 males and 10 females) or adenocarcinoma (219 males and 182 females) of the lung and 183 patients with metastatic lung cancer (82 males and 101 females) who had been aged 40 years or over at the time of surgical resection at the Cancer Institute in Tokyo from 1973 –1991. The tumors which developed in a main or segmental bronchus were classified as central type, and those in a subsegmental or more distal bronchi were categorized as peripheral type. Cases with adenocarcinoma were classified by a pathologist into two histological subtypes, papillary and tubular types, according to the WHO lung carcinoma classification. Risk of squamous cell carcinoma was strongly associated with cigarette smoking for both central (OR (odds ratio) = 10.3 in males and 4.4 in females) and peripheral sites (OR = 10.7 in males and 6.5 in females). There was no significant association between cigarette smoking and adenocarcinoma for any tumor site or histological subtype in both sexes.
Linkage Analysis of BRCA1 in Japanese Breast Cancer Families
Authors: Inoue Rie , Fukutomi Takashi , Ushijima Toshikazu , Matsumoto Yoshiro , Sugimura Takashi , Nagao Minako
Published: August 2005

We examined the involvement of BRCA1, which plays a major role in Western breast cancer families, in Japanese breast cancer families. Eleven families, in which at least three individuals within third degree relatives were affected by breast cancer, were collected. Five of them were early‐onset breast cancer families, in which the average age at diagnosis was less than 45 years, and the other six were late‐onset families. Ovarian cancer was observed in one patient in the early‐onset families. Using seven polymorphic markers on chromosome 17q21, D17S250, ERBB2, THRA1, D17S579, D17S588, GIP and NME1, linkage to BRCA1 was analyzed. Linkage was not detected in any single family. Assuming homogeneity in an inherited component that confines the susceptibility to breast cancer in all families, we summed the LOD scores of all families. The cumulative LOD score obtained was –1.86 for D17S588 at θ= 0.001, indicating no linkage with BRCA1. Since the proportion of families linked to BRCA1 is larger in Western early‐onset breast cancer families than in late‐onset ones, we also summed the LOD scores of five early‐onset families. However, again a negative LOD score was obtained. These results suggest that BRCA1 is not a major breast cancer susceptibility gene in Japanese familial breast cancer.
New Human Oral Squamous Carcinoma Cell Line and Its Tumorigenic Subline Producing Granulocyte Colony‐stimulating Factor
Authors: Matsuo Kou , Ishibashi Yukiko , Kobayashi Ieyoshi , Ozeki Satoru , Ohishi Masamichi , Tange Tsuyoshi , Hirata Jouji , Kiyoshima Tamotsu , Sakai Hidetaka
Published: August 2005

A new human carcinoma cell line, MISK81‐5, was established from a metastatic lymph node of oral squamous cell carcinoma. Immunocytochemical and ultrastructural observations revealed an obvious epithelial origin of the cell line. Chromosome analysis revealed a hypertriploid karyotype with numerical and structural anomalies. MISK81‐5 cells could form a tumor mass in the subcutaneous tissue of recipient BALB/c athymic mice only when coinjected with Matrigel. A stem cell assay revealed that conditioned medium (CM) of MISK81‐5 contained granulocyte colony‐stimulating factor (G‐CSF) or interleukin‐6 activity. Quantitation by ELISA disclosed a higher concentration of G‐CSF in the CM of MISK81‐5 than in the CM of other squamous and gastric carcinoma cell lines. The sMISK, that was derived from MISK81‐5 as a subpopulation of the cell line having higher tumorigenicity, also showed a similar hematopoietic stimulating activity to that of MISK81‐5. These characteristics of the MISK81‐5 cell line and its subpopulation, sMISK will be useful for studying the biological behavior of oral squamous cell carcinomas and its relation to hematopoietic stimulating factors.
Immunohistochemical Analysis of Cyclin D1 Protein in Hematopoietic Neoplasms with Special Reference to Mantle Cell Lymphoma
Authors: Nakamura Shigeo , Seto Masao , Banno Shogo , Suzuki Susumu , Koshikawa Takashi , Kitoh Kuniyoshi , Kagami Yoshitoyo , Ogura Michinori , Yatabe Yasushi , Kojima Masaru , Motoori Tadashi , Takahashi Toshitada , Ueda Ryuzo , Suchi Taizan
Published: August 2005

Immunohistochemical expression of PRAD1/cyclin D1 protein has been investigated in 106 tissue specimens of 104 cases of lymphoma, non‐neoplastic lymphoid disorders and other hematologic malignancies by employing the monoclonal antibody 5D4 with formalin‐fixed paraffin‐embedded sections, using the microwave oven heating method. Positive neoplastic cells were found in 60 (74%) of 81 cases of non‐Hodgkin's lymphoma. The positivity pattern was nuclear in 17 (85%) of 20 cases of mantle cell lymphoma in which cytoplasmic staining was also seen. This pattern of cyclin D1 positivity was in contrast to the negative staining of normal reactive mantle zones. In the other cases, positivity appeared to lie within the cell cytoplasm without nuclear staining, and most of the nodal follicular and diffuse B‐cell lymphomas variously expressed PRAD1/cyclin D1. In contrast, the reaction was absent in a significant number of T‐cell and extranodal B‐cell lymphomas. Immunolocalization of PRAD1/cyclin D1 expression appears to be a useful diagnostic adjunct to discriminate mantle cell lymphoma from other non‐Hodgkin's lymphomas.
Synthetic Oligonucleotides with Certain Palindromes Stimulate Interferon Production of Human Peripheral Blood Lymphocytes in vitro
Authors: Yamamoto Toshiko , Yamamoto Saburo , Kataoka Tetsuro , Komuro Katsutoshi , Kohase Masayoshi , Tokunaga Tohru
Published: August 2005

We studied the ability of synthetic single‐stranded 30‐mer oligodeoxyribonucleotides oligoDNAs) with three different kinds of hexamer palindromic sequence to induce interferon (IFN) production of human peripheral blood lymphocytes (PBL). When PBL was cultured with oligoDNA having a palindrome of AACGTT or GACGTC, IFN activity was detected by bioassay in the culture fluid after 8 h, and the amount of IFN reached the maximum after 18 h. IFN‐α was predominantly produced, and small amounts of IFN‐β and IFN‐γ were also found. OligoDNA with the palindrome ACCGGT had no effect.
Induction of Extrahepatic Biliary Carcinoma by N‐Nitrosobis(2‐oxopropyl)amine in Hamsters Given Cholecystoduodenostomy with Dissection of the Common Duct
Authors: Tajima Yoshitsugu , Eto Toshifumi , Tsunoda Tsukasa , Tomioka Tsutomu , Inoue Keiji , Fukahori Tomohiro , Kanematsu Takashi
Published: August 2005

The methods we used to produce a carcinoma in the extrahepatic bile duct and gallbladder in hamsters are described along with the characteristics of the induced tumors. Female Syrian golden hamsters were first subjected to Cholecystoduodenostomy with dissection of the extrahepatic bile duct on the distal end of the common duct (CDDB) and were, 4 weeks later, treated with weekly subcutaneous injections of N‐nitrosobis(2‐oxopropyl)amine (BOP) at a dose of 10 mg/kg body weight for 9 weeks. The animals were killed at the 12th, 16th and 20th week after the initiation of BOP treatment. Extrahepatic bile duct carcinoma developed in 16%, 24% and 41% and gallbladder carcinoma occurred in 58%, 81% and 82% of the hamsters, respectively, at the corresponding times of killing. The incidences were significantly higher than those in sham‐operated controls (P<0.01). The induced extrahepatic bile duct carcinomas were predominantly of the polypoid type and gallbladder carcinomas were of the papillary type in growth form, being morphologically similar to early stage biliary carcinoma in humans. Immunohistochemical staining using bromodeoxynridine and anti‐bromo‐deoxyuridine monoclonal antibody demonstrated that the CDDB procedure greatly accelerated the cell kinetic activity of the biliary epithelium, and this was considered to be a major factor promoting the development of biliary carcinomas in this hamster model. In conclusion, this new model provides a high incidence of tumor development at the extrahepatic biliary tract and is expected to be useful for clarifying the characteristics of this highly malignant tumor.
State of Adenomatous Polyposis Coli Gene and ras Oncogenes in Japanese Prostate Cancer
Authors: Suzuki Hiroyoshi , Aida Sara , Akimoto Susumu , Igarashi Tatsuo , Yatani Ryuichi , Shimazaki Jun
Published: August 2005

Genetic alterations of ras oncogenes (K‐, H‐ and N‐ras) and adenomatous polyposis coli (APC) gene in tissues of prostate cancer from Japanese patients were examined using PCR‐SSCP (polymerase chain reaction‐single strand conformation polymorphism) analysis and direct sequencing. Tissues from 8 cases of untreated stage B prostate cancer surgically removed and from 10 cases of endocrine therapy‐resistant metastatic disease obtained at autopsy were used in the present study. In four out of 18 cases (22%), ras point mutations were found, two in either codon 12 or 61 of K‐ras and two in either 13 or 61 of H‐ras. These point mutations were detected in one of the stage B cases (13%) and in three of the autopsy cases (30%). All these cases were poorly differentiated adenocarcinoma. In autopsy cases showing ras mutation in cancerous prostate, the same alteration was observed in metastatic tissues. No APC gene mutation was detected in any sample, although polymorphism was found in some cases. These results indicate that ras oncogene mutations are related to the progression of prostate cancer, whereas APC gene alteration is not involved in tumorigenesis and development of this cancer.
Induction Mechanism of Human Blood CD8+T Cell Proliferation and Cytotoxicity by Natural Killer Cell Stimulatory Factor (Interleukin‐12)
Authors: Nabioullin Roustem , Sone Saburo , Nil Akihiko , Haku Takashi , Ogura Takeshi
Published: August 2005

Natural killer cell stimulatory factor (NKSFJIL‐12) has been found to induce cytotoxic activity of human blood T cells. In the present study, the effect of NKSF on induction of cytotoxic CD8+T cells in the presence or absence of monocytes was examined. Highly purified lymphocytes (>99%) and monocytes (>90%) were isolated hy centrifugal elutriation from peripheral blood of normal donors. Then, CD8+cells were isolated with antibody‐bound magnetic beads from purified lymphocytes. The cytotoxicity of CD8+cells was measured by51Cr release assay for 4 h. NKSF enhanced the proliferative response of CD8+cells stimulated with suboptimal concentrations of interleukin‐2 (IL‐2), but rather inhibited their proliferative and cytotoxic responses on stimulation with an optimal concentration of IL‐2. NKSF stimulated CD8+cells to produce interferon 7 (IFNγ) irrespective of the presence of added IL‐2, and this effect was augmented by co‐cultivation with monocytes. Blood monocytes upregulated induction of cytotoxic CD8+cells stimulated with NKSF alone, and this effect was abolished by addition of antibody against IFNγ, but not of antibody against tumor necrosis factora.Induction of NKSF‐inducible cytotoxic CD8+cells was inhibited by addition of transforming growth factor β, but not of IL‐4. These observations suggest thatin situinduction of NKSF‐stimulated cytotoxic CD8+cells may be regulated by complex cytokine networks, depending on the participation of monocytes.
Monoclonal Antibody against PRADl/Cyclin Dl Stains Nuclei of Tumor Cells with Translocation or Amplification at BCL‐1 Locus
Authors: Banno Shogo , Yoshikawa Kazuhiro , Nakamura Shigeo , Yamamoto Kazuhito , Seito Tsutomu , Nitta Masakazu , Takahashi Toshitada , Ueda Ryuzo , Seto Masao
Published: August 2005

Mouse monoclonal antibodies were produced against the bacterial product encoded by human PRAD1/ cyclin Dl gene, which is known to be involved in tumors with translocation or amplification at BCL‐1 locus of 11ql3. The immunizing antigens used were GST‐PRAD1 and T7 gene 10‐PRAD1 fusion products. Four antibodies were reactive with both PRAD1 fusion products and cell lysates of B‐cell tumor cell lines with t(ll;14)(ql3;q32) and a breast cancer cell line with 11ql3 amplification, on immunoblotting. An immunofluorescence study showed that only one of them stained nuclei of cells with 11q13 abnormalities. Since this antibody proved applicable for conventional paraffin‐embedded tissue sections, immunohistologic staining of various lymphoma tissues was performed. Eight of 11 mantle cell lymphomas showed intermediate to strong positivity and 6 of the positive cases demonstrated characteristic staining patterns that were either predominantly nuclear or both nuclear and cytoplasmic. The nuclear staining pattern was not observed with other types of lymphoma and thus may correlate with PRAD1 mRNA overexpression.
Formation and Removal of DNA Adducts in the Liver of Rats Chronically Fed the Food‐borne Carcinogen, 2‐Amino‐3,8‐dimethylimidazo[4,5‐f]quinoxaline
Authors: Hirose Misako , Wakabayashi Keiji , Ochiai Masako , Kushida Hiromi , Sato Hidetaka , Sugimura Takashi , Nagao Minako
Published: August 2005

Effects of chronic administration of 2‐amino‐3,8‐dimethylimidazo[4,5‐f]quinoxaline (MeIQx) at 0.4, 8 and 400 ppm in the diet on DNA adduct formation and removal in the rat liver were examined by the 32P‐postlabeling method. The 0.4 and 8 ppm doses for 40 weeks resulted in time‐dependent increases in MeIQx‐DNA adduct levels until 16 and 8 weeks, respectively, with constant values being maintained thereafter. In the case of a carcinogenic dose (400 ppm) of MeIQx, the adduct levels reached a maximum at week 12, and then gradually decreased. Alteration of metabolism of MeIQx during liver carcinogenesis might be related to this decrease in DNA adduct levels. When MeIQx administration was stopped at week 20, 60–90% of the MeIQx‐DNA adducts formed with the three doses (0.4, 8 and 400 ppm) of MeIQx were removed in a biphasic manner after return to a basal diet, with initial rapid removal followed by a slow change. No difference in the pattern of MeIQx‐DNA adducts was detected on thin layer chromatography at any dose at any time point. Thus, it is suggested that there may be at least two types of damaged DNA, susceptible and resistant to removal of MeIQx‐DNA adducts, after chronic administration of MeIQx.
Genetic Regulation of Development of Thymic Lymphomas Induced by N‐Propyl‐N‐nitrosourea in the Rat
Authors: Ogiu Toshiaki , Fukami Hiroko , Nishimura Mayumi , Matsuyama Mutsushi
Published: August 2005

To clarify the linkage between Hbb and Tls‐1 (thymic lymphoma susceptible‐1) loci and to investigate other loci concerned in thymic lymphomagenesis, the BUF/Mna rat, which is highly sensitive to the lymphomagenic activity of N‐propyl‐N‐nitrosourea (PNU), the WKY/NCrj rat, reported to be resistant, and their cross offspring were subjected to genetic analysis. F1 hybrid and backcross generations were raised from the 2 strains, and 6 genetic markers including Hbb were analyzed in individuals of the backcross generation. However, no linkage between Hbb and Tls‐1 loci could be demonstrated since WKY rats also developed a high incidence of thymic lymphomas in response to PNU. Nevertheless, thymic lymphomas developed more rapidly and reached a larger size in the BUF rats. F1 rats expressed a rather rapid and large tumor growth phenotype, while the [(WKY × BUF) × WKY] backcross generation consisted of rats with either rapidly growing or slowly growing tumors. It was thus concluded that rapid development of thymic lymphomas is determined by a gene, provisionally designated Tls‐3. Analysis of the relationship between 6 genetic markers and development of thymic lymphoma in the backcross generation demonstrated that the Tls‐3 locus is loosely linked to the Gc locus, suggesting a possible location on rat chromosome 14. Tls‐3 may not be identical with Tls‐1 and other genes known to be relevant to thymic tumors, but its relationship with Tls‐2 remains obscure.
A Maternal Risk Factor for Mother‐to‐Child HTLV‐I Transmission: Viral Antigen‐producing Capacities in Culture of Peripheral Blood and Breast Milk Cells
Authors: Yoshinaga Mitsuhiro , Yashiki Shinji , Oki Toshitaka , Fujiyoshi Toshinobu , Nagata Yukihiro , Sonoda Shunro
Published: August 2005

We examined the relationship between productivity of HTLV‐I antigen‐positive cells in cultured peripheral blood mononuclear cells (PBMC) and breast milk mononuclear cells (BMMC) and the incidence of mother‐to‐child transmission of HTLV‐I. Among 61 cases of HTLV‐I carrier mothers, 17 cases were revealed to produce large numbers of HTLV‐I antigen‐positive cells (high HTLV‐I antigen‐producing mothers) whose positive rate was 9.6% in PBMC and 10.2% in BMMC, while the remaining 44 cases produced small numbers of HTLV‐I antigen‐positive cells (low HTLV‐I antigen‐producing mothers) whose positive rate was 0.3% In PBMC and 0.5% in BMMC. The HTLV‐I transmission rate among children born to the high HTLV‐I antigen‐producing mothers was 37.5% (6/16 children from 11 mothers), while that of the low HTLV‐I antigen‐producing mothers was 3.2% (1/31 children from 20 mothers). The transmission rate of HTLV‐I was significantly different between high and low HTLV‐I antigen‐producing mothers (P<0.05). However, there was no positive relationship between anti‐HTLV‐I antibody titers and productivity of HTLV‐I antigen‐positive cells (P=0.11). These results suggested that mother‐to‐child transmission of HTLV‐I might be influenced by a maternally determined factor to produce HTLV‐I antigen‐positive cells in PBMC and BMMC of HTLV‐I carrier mothers.
T‐Cell Receptor Gene Structures of HLA‐A26‐restricted Cytotoxic T Lymphocyte Lines against Human Autologous Pancreatic Adenocarcinoma
Authors: Ueda Daisuke , Sato Noriyuki , Matsuura Akihiro , Sasaki Aya , Takahashi Shuji , Ikeda Hideyuki , Wada Yoshimasa , Kikuchi Kokichi
Published: August 2005

We isolated two cytotoxic T lymphocyte (CTL) lines, which were independently obtained by mixed lymphocyte‐tumor cell culture from tumor‐infiltrating lymphocytes of a patient with pancreatic adenocarcinoma. Both lines behaved identically in all the functional aspects tested and appeared to be HLA‐A26‐restricted. We analyzed their T cell receptor (TCR) gene structures, including V‐(D)‐J junctional sequences, which are unique to each T‐cell clonotype and contribute to TCR diversity. Each line consisted of a clonal T‐cell expressing Vα18 and Vβ7. The α chain gene was composed of Vα18/ JαF/Cα and the, β‐chain gene, of Vβ7.1/Dβ/Jβ1.4/Cβ2. The sequences were all in‐frame and therefore should yield functional transcripts. The junctional sequences were identical between the two lines. These data suggested that the two CTL clones having the same CDR3 had descended from a common precursor lymphocyte. The clonal expansion of CTL lines with the identical CDR3 implies that they are directed against the same tumor antigen, which seemed to be immunologically dominant in the specific interaction between the CTL and the autologous pancreatic adenocarcinoma.
Interventional Trial for Colorectal Cancer Prevention in Osaka: An Introduction to the Protocol
Authors: Ishikawa Hideki , Akedo Ikuko , Suzuki Takaichiro , Otani Toru , Sobue Tomotaka
Published: August 2005

We established a protocol for an interventional randomized controlled trial for prevention of colorectal cancer by attaching special importance to feasibility. The subjects were patients with multiple colorectal tumors. Two regimens were formulated for prevention of colorectal cancer. One was dietary guidance alone (Regimen I), and the other was dietary guidance plus eating wheat bran biscuits (Regimen II). The main end points of the trial were examinations for recurrence of colorectal tumors after 2 and 4 years. The target number of patients was 200 in total, i.e. 100 for each group. During the 18 months from the beginning of recruiting of subjects (up to November 1994), 28 (97%) of the 29 patients recruited for Regimen I and 32 (97%) of the 33 patients recruited for Regimen II agreed to participate in the trial. The trial is progressing well.
Activation of Protein Kinase C by Mycobacterial Cord Factor, Trehalose 6‐Monomycolate, Resulting in Tumor Necrosis Factor‐α Release in Mouse Lung Tissues
Authors: Sueoka Eisaburo , Nishiwaki Shinji , Okabe Sachiko , Iida Naoyuki , Suganuma Masami , Yano Ikuya , Aoki Kunio , Fujiki Hirota
Published: August 2005

Cord factors are mycoloyl glycolipids in cell walls of bacteria belonging to Actinomycetales, such as Mycobacterium, Nocardia and Rhodococcus. They induce granuloma formation in the lung and interstitial pneumonitis, associated with production of macrophage‐derived cytokines. We studied how cord factors induce biological activities in the cells. Cord factors isolated from M. tuberculosis, trehalose 6‐monomycolate (mTMM) and trehalose 6,6′‐dimycolate (mTDM), enhanced protein kinase C (PKC) activation in the presence of phosphatidylserine (PtdSer), diacylglycerol and Ca2+, and mTMM activated PKCα more strongly than PKCβ or γ under the same assay conditions. Kinetic studies of mTMM in response to PKC activation revealed that mTMM increased the apparent affinity of PKC to Ca2+ in the presence of both PtdSer and diolein. Although this is similar to observations with unsaturated fatty acids, such as arachidonic acid, mTMM was synergistic with PtdSer for PKC activation, but arachidonic acid was not. mTMM was also different as regards PKC activation, as phorbol ester was. A single i.p. administration of mTMM to mouse induced tumor necrosis factor‐α (TNF‐α) in serum and in the lung, which is a unique target tissue of cord factors. Based on our recent finding that TNF‐α is an endogenous tumor promoter, the correlation between lung cancer and pulmonary tuberculosis is discussed.
Dipyridamole Combined with Tumor Necrosis Factor‐α Enhances Inhibition of Proliferation in Human Tumor Cell Lines
Authors: Suzuki Nobuo , Sekiya Souei , Sugano Isamu , Kojima Takayuki , Yamamori Hideo , Takakubo Yoshiaki
Published: August 2005

In the search for cytokines whose antiproliferative action could be enhanced by combination with dipyridamole, 2,6‐bis(diethanolamino)‐4,8‐dipiperidinopyrimido[5,4,d]pyrimidine, the combination of tumor necrosis factor‐α (TNF‐α) with this agent was evaluated in various human tumor cell lines. Inhibition of the proliferation of human melanoma cell lines MM‐1CB and HMV‐1 by TNF‐α (1–102 U/ml) was enhanced in culture dishes by combination treatment with dipyridamole (0.1–10 μM). The enhancement effect was also detected in other tumor cell lines: T9S (glioma), SCC‐1CB (squamous cell carcinoma), HAC‐2 (ovarian clear‐cell carcinoma), HLE (hepatoma), HEC‐1 (endometrial adenocarcinoma) and HOC‐21 (ovarian serous cystadenocarcinoma). The incorporation of [14C]amino acids and [3H]nridine into acid‐insoluble cell materials in the combination‐treated cells was not significantly different from that in cells treated with TNF‐α or dipyridamole. However, the incorporation of [3H]thymidine was specifically inhibited in all cell lines examined after more than 12 h of the TNF‐α and dipyridamole combination treatment, although neither agent alone inhibited this incorporation. On the other band, the growth of tumors induced by the injection of MM‐1CB and HMV‐1 cells into nude mice was more markedly inhibited by the subcutaneous administration of TNF‐α in combination with orally administered dipyridamole than by either agent alone. The results presented suggested that dipyridamole is beneficial in assuring the effectiveness of anti‐cancer cytokine therapy.
Serum Antibody against Unfused Recombinant E7 Protein of Human Papillomavirus Type 16 in Cervical Cancer Patients
Authors: Fujii Takuma , Matsushima Youko , Yajima Masazumi , Sugimura Takashi , Terada Masaaki
Published: August 2005

Sera were examined for the presence of antibody against E7 protein of human papillomavirus type 16 (HPV‐16) by Western blot analysis using the bacterially derived unfused protein. The occurrence rates of anti‐E7 antibody against HPV‐16 were 14.1% (10/71) in cervical cancer patients, 0% (0/48) in cervical intraepithelial neoplasia patients, and 0% (0/41) in female non‐malignant patients. Three patients (one with endometrial cancer, one with breast cancer, and one male patient with colon polyp) out of 115 patients with tumors in organs other than the cervix, had antibody against E7 protein of HPV‐16. The serum antibody, once positive, could be detected for a long time after surgical removal of the cancers in all cases that could be followed up. HPV‐16 DNA could be detected in 50% (13/26) of cervical cancer patients. Sixty‐nine percent (9/13) of patients with HPV‐16 DNA in cancers had the antibody and all the patients with stages II, III, and IV cervical cancer (8/8) harboring HPV‐16 DNA showed the presence of the antibody against E7 protein of HPV‐16. In contrast, only 20% (1/5) of cervical cancer patients with stage Ia or Ib harboring HPV‐16 DNA showed positive for the anti‐E7 antibody in sera. These findings suggest that the presence of anti‐E7 antibody in serum depends on the staging of cervical cancer and extent of HPV infection.
Inhibition of Pulmonary Metastases and Tumor Cell Invasion in Experimental Tumors by Sodium d‐Glucaro‐δ‐lactam (ND2001)
Authors: Tsuruoka Tsutomu , Fukuyasu Harumi , Azetaka Masayuki , Iizuka Yumiko , Inouye Shigeharu , Hosokawa Masuo , Kobayashi Hiroshi
Published: August 2005

Sodium d‐glucaro‐δ‐lactam (ND2001) inhibited spontaneous pulmonary metastases of the highly metastatic B16 melanoma variant with a maximal inhibition rate of 99.5%, and 6 of 7 animals remained metastasis‐free. Likewise, ND2001 inhibited the spontaneous pulmonary metastases of both Lewis lung carcinoma (3LL) with a rate of 98.0% (3 of 5 animals remaining metastasis‐free) and rat KDH‐8 liver carcinoma with a rate of 82.5% (3 of 7 animals remaining metastasis‐free), although it was unable to inhibit the metastases of mouse BMT‐11 fibrosarcoma and rat SST‐2 breast carcinoma. Pretreatment with ND2001 in vitro inhibited the pulmonary metastases of the B16 variant and 3LL cells, which indicates direct action upon the cancer cells. When the invasive activity of cancer cells was measured by the Boyden chamber method, the number of invading B16 variant or 3LL cells was reduced with maximal inhibition rates of 93.0% or 89.9%, respectively, but pretreatment with ND2001 failed to reduce the invasive activity of BMT‐11 or SST‐2 cells. ND2001 showed neither cytocidal nor antitumor activity. These results suggest that ND2001 inhibited pulmonary metastases at the invasive step into the basement membrane by directly changing some property of the tumor cells.
p53 Mutations Occur in Clinical, but Not Latent, Human Prostate Carcinoma
Authors: Konishi Noboru , Hiasa Yoshio , Hayashi Isao , Matsuda Hirofumi , Tsuzuki Toshihide , Ming Tao , Kitahori Yoshiteru , Shiraishi Taizo , Yatani Ryuichi , Shimazaki Jun
Published: August 2005

To elucidate the role of the p53 tumor suppressor gene in prostate tumorigenesis, we probed for mutations in latent and clinical prostate cancers using single strand conformation polymorphism (SSCP) and restriction fragment length polymorphism (RFLP) analysis in combination with direct gene sequencing and immunohistochemical methodologies. Fifteen cases of subclinical and 32 cases of clinical carcinoma, the latter graded in stages A through D, were available for study. While p53 point mutations were detected in only 5 of 32 (16%) clinical cancers, no mutations were detected in latent disease. Of the carcinomas in stages B, C and D, 15% (2/13), 29% (2/7) and 9% (1/11) were positive for p53 mutations, respectively. Although no specific mutational patterns were observed, the aberrations found were predominantly single base missense substitutions. The data suggest not only an association of p53 mutation and progression of clinical prostate cancer, but also imply that some other mechanism(s) are at work in latent carcinoma.
Reinforced Cytotoxicity of Lymphokine‐activated Killer Cells toward Glioma Cells by Transfection of the Killer Cells with the γ‐Interferon Gene
Authors: Mizuno Masaaki , Yoshida Jun , Takaoka Toru , Sugita Kenichiro
Published: August 2005

Lymphokine‐activated killer (LAK) cells generated from peripheral blood lymphocytes incubated with recombinant interleukin‐2 were transfected with the human γ‐interferon (HuIFN‐γ) gene by means of liposomes having a positive charge on their surface. The cells secreted significant amounts of HuIFN‐γ (reaching more than 5 U/ml) into the culture medium. The HuIFN‐γ produced by the cells induced intercellular adhesion molecule‐1 (ICAM‐1) and enhanced the expression of Fas antigen on the surface of human glioma cells. Also, LAK cells transfected with HuIFN‐γ gene exhibited reinforcement of cytotoxicity toward human glioma cell lines (U251‐MG and SK‐MG‐1). Furthermore, the reinforcement was significantly quenched by anti‐ICAM‐1 and/or anti‐TNF‐α monoclonal antibody.
A Pharmacokinetic and Pharmacodynamic Analysis of CPT‐11 and Its Active Metabolite SN‐38
Authors: Sasaki Yasutsuna , Hakusui Hideo , Mizuno Shoichi , Morita Masashige , Miya Toshimichi , Eguchi Kenji , Shinkai Tetsu , Tamura Tomohide , Ohe Yuichiro , Saijo Nagahiro
Published: August 2005

In the present study, an attempt was made to determine the precise pharmacokinetics of 7‐ethyl‐10‐[4‐(1‐piperidino)‐1‐piperidino]carbonyloxycamptothecin (CPT‐11) and its active metabolite, 7‐ethyl‐10‐hydroxycamptothecin (SN‐38). The relationship between pharmacokinetic parameters and pharmacodynamic effects was also investigated to elucidate the cause of interpatient variation in side effects. Thirty‐six patients entered the study. CPT‐11, 100 mg/m2, was administered by IV infusion over 90 min weekly for four consecutive weeks. The major dose‐limiting toxicities were leukopenia and diarrhea. There was a positive correlation between the area under the concentration‐time curve (AUC) of CPT‐11 and percent decrease of WBC (r=0.559). On the other hand, episodes of diarrhea had a better correlation with the AUC of SN‐38 (r=0.606) than that of CPT‐11 (r=0.408). Multivariate analysis revealed that the AUC of SN‐38, AUC of CPT‐11 and indocyanine green retention test were significant variables for the incidence of diarrhea and that both performance status and AUC of CPT‐11 were significant variables for percent decrease of WBC. The large interpatient variability of the degree of leukopenia and diarrhea is due to a great plasma pharmacokinetic variation in CPT‐11 or SN‐38. The AUCs of CPT‐11 and SN‐38 obtained from the first administration of CPT‐11 correlate with toxicities, but it is impossible to predict severe side effects before the administration of CPT‐11 at the present time.
Cloning of the Rat Homologue of the von Hippel‐Lindau Tumor Suppressor Gene and Its Non‐somatic Mutation in Rat Renal Cell Carcinomas
Authors: Kikuchi Yasushi , Kobayashi Etsuko , Nishizawa Masae , Hamazaki Shuji , Okada Shigeru , Hino Okio
Published: August 2005

Recently, von Hippel‐Lindau (VHL) gene mutations were detected in non‐inherited, sporadic human renal cell carcinomas (RCs) at a high frequency. In order to determine whether or not the VHL gene is also a critical gene in rat RCs, we cloned and sequenced the rat homologue of human VHL gene and searched for mutations of the VHL gene in rat RCs. Mutations in the VHL gene were not detected in spontaneous RCs of the Eker rat model or in ferric nitrilotriacetate‐induced rat RCs using the polymerase chain reaction‐single strand conformation polymorphism (PCR‐SSCP) method. These data indicate that mutation of the VHL tumor suppressor gene is not an event in rat renal carcinogenesis, at least in our present systems.
Prognostic Value of p53 Protein Accumulation in Cancer Cell Nuclei in Adenocarcinoma of the Uterine Cervix
Authors: Tsuda Hitoshi , Jiko Kumiko , Tsugane Shoichiro , Yajima Masazumi , Yamada Takuro , Tanemura Kenjiro , Tsunematsu Ryuichiro , Ohmi Kazuo , Sonoda Takahiko , Hirohashi Setsuo
Published: August 2005

Adenocarcinoma of the uterine cervix (CxAd) is one of the most distressing malignancies of the female reproductive system because of its tendency to spread aggressively and to be resistant to radiation and systemic therapies. To clarify the prognostic significance of p53 alteration in CxAd, we immunohistochemically examined the incidence of p53 nuclear accumulation, which is considered to be mostly parallel with p53 gene mutation, and its association with clinicopathological parameters in 26 patients with CxAd. The overall incidence of p53 nuclear accumulation was 46% (12 of 26), being higher in groups with clinically advanced disease, higher degrees of cellular atypia, and deeper myometrial invasion, but significantly lower in patients with integration of human papillomavirus (HPV) type 16 or 18 DNA. Nuclear p53 immunoreactivity as well as lymph node status, depth of invasion and the absence of HPV‐DNA integration were significant indicators of a poor prognosis. Examination of p53 nuclear accumulation could be applied to biopsy material, and would be of practical assistance in predicting the prognosis of CxAd both preoperatively and postoperatively.
Expression of Sulfomucins in Normal Mucosae, Colorectal Adenocarcinomas and Metastases
Authors: Matsushita Yoshifumi , Yamamoto Noriko , Shirahama Hiroshi , Tanaka Sadao , Yonezawa Suguru , Yamori Takao , Irimura Tatsuro , Sato Eiichi
Published: August 2005

We have examined the expression of specific mucin antigens in tissue sections from 92 cases of colorectal carcinoma, using sulfomucin‐specific monoclonal antibody (MAb) 91.9H. The expression of sulfomucins was high in normal mucosae and much lower in primary colorectal carcinoma, in metastatic lesions in lymph nodes or in liver. The intracellular localization of sulfomucins was also different among these tissues. In normal mucosae, MAb 91.9H binding was seen in the supranuclear area, presumably Golgi complexes, the luminal surface, and secretory products. In primary colorectal carcinomas and in their metastatic lesions, MAb 91.9H was preferentially localized in the cell surface and substances attached to the luminal surface of glandular structures. Analysis of the lysates of normal and tumor tissues showed that very‐high‐molecular‐weight components contained the antigenic epitopes. The intensity of MAb 91.9H binding was lower in tumors at advanced stages than in tumors at early stages. These high‐molecular‐weight components were apparently reactive with MAb FH6 specific for sialyl‐Lex (s‐Lex) structures. Histological specimens with low levels of MAb 91.9H reactivity often exhibited relatively high levels of MAb FH6 reactivity. These two mucins may have reversed expression during carcinogenesis and carcinoma progression, and this change may be related to metastatic potential.
Frequent Development of Murine T‐Cell Lymphomas with TcRα/β+, CD4‐/8 Phenotype after Implantation of Human Inflammatory Breast Cancer Cells in BALB/c Nude Mice
Authors: Wakasugi Hiro , Koyama Kotaro , Gyotoku Motomichi , Yoshimoto Masataka , Hirohashi Setsuo , Sugimura Takashi , Terada Masaaki
Published: August 2005

Tumors developed quite frequently in some of the visceral organs, including spleen and liver, in BALB/c nude mice upon subcutaneously xenografting surgical specimens from five different inflammatory breast cancer patients. All of these tumors developed within two and a half months to one year after the subcutaneous inoculation of surgical specimens. From these tumors, five independent transplantable tumors, including tMK‐2, tHK‐1, tYK‐1, tYK‐2 and tTY‐1 have been established. Chromosome analysis, morphologic studies by light and electron microscopy and phenotype analysis indicated that these tumors are of mouse origin. The tMK‐2 tumor was highly metastatic to the spleen and liver when it was subcutaneously transplanted into the right scapular region. In addition, the region where the tMK‐2 tumor cells were subcutaneously inoculated showed an apparently inflammatory process represented by erythema. After subcutaneous inoculation into the right scapular region, tHK‐1, tYK‐1,2, and tTY‐1 tumors also metastasized to some of the visceral organs, including spleen and liver. From these tumors, in vitro cell lines were established. The cells grew in a stromal‐cell dependent manner under in vitro culture conditions. The cells were again tumorigenic at the inoculated region and metastasized to various organs, including liver and spleen, of BALB/c nude mice. Histological examination revealed that the tumors showed features of malignant lymphoma. Phenotypically, these five tumors expressed early T lymphocyte markers as revealed by anti‐mouse anti‐TcR4aL/β, anti‐CD3, CD4 and CDS monoclonal antibodies. To our knowledge, these cell lines are the first T‐cell lines showing the phenotype of extrathymically differentiated T‐cells in the liver.
Inhibition of 1,2‐Dimethylhydrazine‐induced Oxidative DNA Damage by Green Tea Extract in Rat
Authors: Inagake Masao , Yamane Tetsuro , Kitao Yoshitaka , Oya Kazuhiko , Matsuraoto Hirohiko , Kikuoka Norikazu , Nakatani Hirohisa , Takahashi Toshio , Nishimura Hiroshi , Iwashiraa Akio
Published: August 2005

Following subcutaneous injection of 1,2‐dimethylhydrazine (DMH), which is carcinogenic to rat colon and liver, to Sprague‐Dawley rats, a significant increase of 8‐hydroxydeoxyguanosine (8‐OHdG) was observed in the DNA of colonic mucosa and liver. The 8‐OHdG formation reached the maximal level at about 24 h after the DMH injection. On the other hand, no increase of 8‐OHdG was observed in the DNA of the kidney. Drinking green tea extract (GTE) for ten days prior to the DMH injection significantly inhibited the formation of 8‐OHdG in the colon. These findings demonstrate that DMH causes oxidative damage to the DNA of its target organ, and that GTE protects colonic mucosa from this oxidative damage.
Interleukin‐12 Augments the Generation of Autologous Tumor‐reactive CD8+ Cytotoxic T Lymphocytes from Tumor‐infiltrating Lymphocytes
Authors: Kuge Soichi , Watanabe Kazuhito , Makino Koji , Tokuda Yutaka , Mitomi Toshio , Kawamura Nobuo , Habu Sonoko , Nishimura and Takashi
Published: August 2005

Human tumor‐infiltrating lymphocytes (TIL) were obtained from breast cancer, renal cancer or neuroblastoma to investigate the generation of autologous tumor‐reactive CD8+ cytotoxic T lymphocytes (CTL). When TIL were cultured with interleukin (IL)‐2 (100 U/ml), the growth of TIL peaked around 8–10 days after the initiation of culture. In contrast, the proliferation of TIL cultured with IL‐2 plus IL‐12 peaked around 4–5 days after culture and tumor cells rapidly disappeared from the culture. To determine the generation of autologous tumor‐reactive CD8+ CTL, TIL‐derived CD8+ T cells were separated by FACStar. Both IL‐2‐activated and IL‐2 plus IL‐12‐activated TIL‐CD8+ T cells showed the same level of lymphokine‐activated killer activity against a variety of tumor cells. However, TIL‐CD8+ T cells activated with IL‐2 plus IL‐12 revealed greatly augmented cytotoxicity against autologous tumor cells compared with that induced by IL‐2 alone. The autologous tumor cell‐killing activity of TIL‐CD8+ CTL was significantly inhibited by the addition of F(ab)2 anti‐CD3 monoclonal antibody, indicating that these CTL recognize autologous tumor antigen through T cell receptor. These results imply that IL‐12 is a novel cytokine which facilitates the generation of autologous tumor‐reactive CD8+ CTL from TIL.