bluuebunny/crossref_metadata_2025 huggingface dataset:
In vivo Effects of a Histone Deacetylase Inhibitor, FK228, on Human Acute Promyelocytic Leukemia in NOD/Shi‐scid/scid Mice
Authors: Kosugi Hiroshi , Ito Masafumi , Yamamoto Yukiya , Towatari Masayuki , Ito Mamoru , Ueda Ryuzo , Saito Hidehiko , Naoe Tomoki
Published: August 2005

Histone acetylation and deacetylation are closely linked to transcriptional activation and repression, respectively. In acute promyelocytic leukemia (APL), histone deacetylase inhibitors (HDACIs) have a synergistic effect with all‐iraws retinoic acid (ATRA) in vitro to induce differentiation. Here we report in vitro and in vivo effects of a HDACI, FK228 (formerly FR901228 or depsipeptide), on the human APL cell line NB4. FK228 had a strong and irreversible cytotoxicity compared with another HDACI, trichostatin A. In vivo administration of ATRA or FK228 alone partly inhibited the growth of established tumors of NB4 subcutaneously transplanted in NOD/Shi‐scid/scid mice, and the combination was synergistically effective. Histopathological examination revealed that the combination induced apoptosis and differentiation as well as histone acetylation. Intravenous injection of NB4 in NOD/Shi‐scid/scid mice followed by combination treatment significantly prevented leukemia death, whereas single administration did not. These findings suggest that FK228 is a promising agent to enhance ATRA‐sensitivity in the treatment of APL.
Different Susceptibility of Each L‐myc Genotype to Esophageal Cancer Risk Factors
Authors: Kumimoto Hiroshi , Hamajima Nobuyuki , Nishizawa Kimiko , Nishimoto Yoshio , Matsuo Keitaro , Harada Hideki , Shinoda Masayuki , Hatooka Shunzo , Ishizaki Kanji
Published: August 2005

To understand the relationship between the L‐myc genotypes and esophageal cancer risk, a polynierase chain reaction‐based restriction fragment length polymorphism analysis was performed on 91 Japanese patients with esophageal cancer and 241 non‐cancer outpatients. No significant difference in the distribution of genotypes was observed between patients and controls; 18.7% LL genotype, 56.0% LS and 25.3% SS among patients, and 24.5%, 55.6% and 19.9%, respectively, among controls. Frequency of the s‐allele in patients (0.533) was slightly higher than in controls (0.477), but the difference was not statistically significant. However, the odds ratios (ORs) for smoking or heavy drinking were markedly higher in SS and LS genotypes than in LL genotype; age‐sex‐adjusted ORs for smoking was 7.57 in the SS genotype, 6.40 in the LS genotype and 1.77 in the LL genotype. Age‐sex‐adjusted ORs for heavy drinking were 19.78, 18.20 and 7.40, respectively. The age‐sex‐adjusted ORs for both factors combined were 12.77, 18.45 and 1.44, respectively. These results suggested that the L‐myc polymorphism might modify the effects of lifestyle factors on esophageal cancer risk.
Expression of Drug Resistance Genes in VP‐16 and mAMSA‐selected Human Carcinoma Cells
Authors: Matsumoto Yoshihito , Takano Hiroshi , Kunishio Katsuzo , Nagao Seigo , Fojo Tito
Published: August 2005

The cell lines described in the present study were isolated as part of an effort to understand resistance to topoisomerase (topo) II inhibitors. To that end, 50 sublines were isolated from four human breast cancer cell lines, i.e., MCF‐7, T47D, MDA‐MB‐231, and ZR‐75B. As an initial step, a concentration that would be lethal to the majority of cells (IC99) was selected for both VP‐16 and mAMSA, for each cell line. The identification of an increasing number of putative drug resistance‐related proteins provided the opportunity to examine expression of the corresponding genes in the selected cell lines. Northern blot analysis revealed different responses to the selecting agents in the different cell lines. Previous studies examining expression of multidrug resistance (MDR)‐l in resistant cell lines had found undetectable levels in all cells. In the ZR‐75B sublines, increased expression of MDR‐associated protein (MRP) and canalicular multispecific organic anion transporter (cMOAT) was observed, and when the relative levels of overexpression were compared, a high correlation was found. In contrast, increased expression of MRP was observed in some of the MDA‐MB‐231 sublines, without a concomitant increase in cMOAT expression. Finally, in both T47D and MCF‐7 sublines, increased expression of cMOAT or MRP was observed infrequently, and where it occurred, was of a much smaller magnitude. In the analysis of expression of MRP, the highest levels were found in the ZR‐75B and MDA‐MB‐231 sublines, with lower levels in the MCF‐7 and T47D clones. Similarly, differences in the expression of topo IIα were observed among the sublines. Although the differences in expression appear to depend on the parental cell line from which the resistant sublines were derived, a strong correlation was observed between the expression of MRP and the levels of topo IIα. Cell lines with low levels of MRP had lower levels of topo IIα, while those with high levels of MRP maintained higher levels of topo IIα. While a reduced topo IIα level was common, there did not appear to be a compensating increase in the expression of topo IIβ or topo I or casein kinase (CK) IIα in any of the cell lines. While the possibility that such compensation could occur has been discussed and even reported in some cell lines, such an adaptation was not observed in the present study, suggesting that it is not common.
Deletion Mutants of Human Deoxycytidine Kinase mRNA in Cells Resistant to Antitumor Cytosine Nucleosides
Authors: Obata Tohru , Endo Yoshio , Tanaka Motohiro , Uchida Hiroyuki , Matsuda Akira , Sasaki Takuma
Published: August 2005

We studied mutational events in deoxycytidine (dCyd) kinase mRNA expression, focusing on aberrant dCyd kinase mRNA, which has been frequently observed in established cell lines resistant to antitumor dCyd nucleoside analogues such as 1‐β‐D‐arabinofuranosyl cytosine (Ara‐C), gemcita‐bine (dFdC) and 2′‐C‐cyano‐2′‐deoxy‐l‐β‐D‐arabinofuranosylcytosine (CNDAC). We describe here the expression of aberrant dCyd kinase mRNAs identified as splicing mutants. These mutants included deletions of the fifth exon in CNDAC‐resistant cells (originating from HT‐1080 cells), of the third exon in Ara‐C‐resistant cells (originating from SK‐MEL‐28 cells) and of the fourth exon in 2′‐deoxy‐2′‐methylidenecytidine (DMDC)‐resistant cells (originating from SK‐MEL‐28 cells). Various nucleoside‐resistant cells originating from the same parental HT‐1080 cells were established. The resulting cells expressed the same mRNA with deletion of the fifth exon, and the location of splicing was independent of the type of nucleosides used for the establishment of resistant cells. The deletion of the fifth exon in dCyd kinase seems to be a target for acquisition of resistance to antitumor cytosine nucleosides. However, distinct mutations in the dCyd kinase gene seem to be associated with acquisition of resistance to different antitumor cytosine nucleosides.
Decrease in Risk of Lung Cancer Death in Males after Smoking Cessation by Age at Quitting: Findings from the JACC Study
Authors: Wakai Kenji , Seki Nao , Tamakoshi Akiko , Kondo Takaaki , Nishino Yoshikazu , Ito Yoshinori , Suzuki Koji , Ozasa Kotaro , Watanabe Yoshiyuki , Ohno Yoshiyuki ,   for the JACC Study Group
Published: August 2005

To evaluate the impact of smoking cessation in individuals and populations, we examined the decrease in risk of lung cancer death in male ex‐smokers by age at quitting in the Japan Collaborative Cohort Study for Evaluation of Cancer Risk Sponsored by Monbusho (JACC Study), which was initiated from 1988 to 1990 in Japan. For simplicity, subjects were limited to male non‐smokers, and former/current smokers who started smoking at ages 18‐22, and 33 654 men aged 40‐79 years were included. We modeled the mortality rates in non‐smokers and current smokers, and compared the rates in ex‐smokers with those expected from the model if they had continued smoking. During the mean follow‐up of 8.0 years, 341 men died from lung cancer. The mortality rate ratio for current smokers, compared to non‐smokers, was 5.16, and those for ex‐smokers who had quit smoking 0‐4, 5‐9,10‐14,15‐19 and >20 years before were 4.84, 3.19, 2.03,1.29 and 0.99, respectively. The functions of 3.20×l0‐7×(age)45and 1.96×lO‐5×(age‐29.6)4.5fitted the observed mortality rates (per 100 000 person‐years) in non‐smokers and continuing smokers, respectively. A greater decrease in lung cancer mortality was estimated among those who quit smoking at younger ages. Stopping smoking earlier in life appears preferable to keep the individual risk low. The absolute rate, however, substantially decreased after smoking cessation even in those who quit at ages 60‐69, reflecting the high mortality rate among continuing smokers in the elderly.
Role of Atp7b Gene in Spontaneous and N‐Diethylnitrosamine‐induced Carcinogenesis in a New Congenic Strain, WKAH.C‐Atp7b Rats
Authors: Minami Takanori , Kaneda Shinya , Otsuka Toshihiro , Jiao Zhongxian , Suzuki Yasuo , Yamada Takahisa , Matsumoto Kozo , Izumi Keisuke
Published: August 2005

To examine whether Long‐Evans Cinnamon (LEC) rats, a mutant rat model of Wilson's disease, have a susceptibility gene(s) to hepatocarcinogenesis in addition to the causative gene, Atp7b, we established a new congenic strain, WKAH.C‐Atp7b rats, in which the Atp7b gene of the LEG rats is inserted into the normal Wistar‐King Aptekman Hokkaido (WKAH) background. Hepatocellular tumors developed spontaneously in both sexes of WKAH.C‐Atp7b rats, their incidence being slightly lower than that in LEG rats. Incidences of spontaneous liver tumors in LEG, WKAH.C‐Atp7b and WKAH rats correlated with hepatic copper and iron concentrations. Medium‐term liver bioassay showed that LEG rats were more susceptible to the induction of glutathione S‐transferase placental form‐positive preneoplastic foci than WKAH.C‐Atp7b rats, and WKAH.C‐Atp7b rats were more susceptible than WKAH rats. In an N‐diethylnitrosamine (DEN)‐induced long‐term carcinogenicity study, 1) LEC rats were similarly or rather less susceptible to hepatocellular tumors than WKAH.C‐Atp7b and WKAH rats, indicating that the progression of the preneoplastic foci to liver cancer in LEG rats was worse than that in WKAH.C‐Atp7b and WKAH rats, 2) the incidences of kidney tumors in LEG and WKAH.C‐Atp7b rats were higher than that in WKAH rats and high copper concentrations in the kidneys were observed in LEG and WKAH.C‐Atp7b rats, 3) LEC rats were resistant to lung carcinogenesis. These data indicate that the susceptibility of LEG rats to liver and kidney carcinogenesis could be explained by Atp7b gene mutation and that the susceptibility to lung carcinogenesis is controlled by gene(s) other than Atp7b.
Anticancer Drug‐mediated Induction of Multidrug Resistance‐associated Genes and Protein Kinase C Isozymes in the T‐Lymphoblastoid Cell Line CCRF‐CEM and in Blasts from Patients with Acute Lymphoblastic Leukemias
Authors: Beck James F. , Brügger Dorothee , Brischwein Klaus , Liu Chao , Bader Peter , Niethammer Dietrich , Gekeler Volker
Published: August 2005

The major determinants mediating drug resistance in acute lymphoblastic leukemias (ALL) unresponsive to chemotherapy, are still unclear. For example, it is still unknown whether selection or induction processes are responsible for drug resistance here or whether protein kinase C (PKC) isozymes contribute to the resistant phenotype. Therefore, inducibility of resistance factors or PKC isozymes genes was examined in CCRF‐CEM cells treated with diverse anticancer drugs‐ adriamycin, camptothecin, etoposide or vincristine‐at sublethal concentrations for 24 h. MDR1, MRP1, LRP and PKC isozyme α, β1, β2, ε, ι, η, θ, ζ gene expression was determined by cDNA‐PCR. We found significant dose‐dependent, mostly combined, induction of the MDR1, MRP1 and LRP genes. Significantly enhanced gene expression of the majority of PKC isozyme genes was found after treatment with camptothecin. PKCζ was upregulated throughout by each anticancer drug applied in this setting. A series of selected CCRF‐CEM‐derived multidrug resistance (MDR) sublines also showed enhanced expression of the PKC isozymes compared to the parental cell line. MDR1 and PKCη gene expression levels were correlated highly significantly. Blasts from two patients with ALL during the first week of monotherapy with steroids revealed combined induction of the MDR1, multidrug resistance‐associated protein 1 (MRP1), lung cancer resistance‐related protein (LRP) and most PKC isozymes, predominantly PKCζ. Another patient with T‐ALL, who failed to respond to four months of intensive chemotherapy, showed an enhanced MRP1 gene expression combined with markedly overexpression of PKCη and PKCθ. Furthermore, the camptothecin and etoposide‐mediated induction of resistance factors in the CCRF‐CEM cell line could be suppressed by staurosporine, a rather unspecific inhibitor of protein kinases. However, selective inhibitors of PKC isozymes (bisindolylmaleimide GÖ 6850, indolocarbazole GÖ 6976) produced no significant effects here. Therefore, the PKC isozymes η, θ and ζ are of interest as potential targets to overcome drug resistance in ALL.
Increased Apoptosis Associated with Depressed Type of Early Intestinal Gastric Cancer
Authors: Masutani Manabu , Suzuki Jun‐ichi , Matsuda Tomoki , Dochin Akiharu , Sadaoka Kuniaki , Nomura Akiyoshi , Ohira Koji , Takahashi Kohei , Yamazaki Koichi , Dosaka‐Akita Hirotoshi , Nishimura Masaharu , Kawakami Yoshikazu
Published: August 2005

Early gastric cancer can be macroscopically classified into elevated and depressed types. To clarify the relationship between macroscopic appearance of early gastric cancer and apoptosis or cell proliferation, formalin‐fixed paraffin‐embedded tissue specimens of 44 intestinal‐type early gastric cancers were investigated by the TUNEL method and immunohistochemical techniques. Diffuse type was excluded in this study. When tissue sections of gastric cancer were vertically classified into the 3 compartments of laminar, intermediate and basal, the apoptosis index (%) was significantly higher in the basal compartment of depressed type (1.76±2.04, mean±SD) than in the basal compartment of elevated type (0.63±0.81, P=0.01). In depressed type, the apoptosis index (%) was significantly higher in the basal compartment than in the luminar compartment (0.76 ±0.85, P=0.03). Apoptosis‐inducing protein, Bax, was expressed more in each of the compartments of depressed type than in those of elevated type, while there were no significant differences in expression of anti‐apoptotic protein, Bcl‐2, between the two types. Moreover, the apoptosis index (%) of Bax‐positive gastric cancer was significantly higher in the basal compartment (P=0.03), compared to that of Bax‐negative gastric cancer, while there were no significant differences in apoptosis index (%) in any compartment between Bcl‐2‐positive and Bcl‐2‐negative gastric cancers. There were no significant differences in Ki‐67 expression, either between the two types, or among the compartments of depressed type. These results indicate that increased apoptosis with excessive expression of Bax in the basal compartment is involved in the morphogenesis of the depressed type in intestinal‐type early gastric cancer.
p53 and H‐ras Mutations and Microsatellite Instability in Renal Pelvic Carcinomas of NON/Shi Mice Treated with N‐Butyl‐N‐(4‐hydroxybutyl)‐nitrosamine: Different Genetic Alteration from Urinary Bladder Carcinoma
Authors: Gen Hiroyuki , Yamamoto Shinji , Morimura Keiichirou , Min Wei , Mitsuhashi Makoto , Murai Takashi , Mori Satoru , Hosono Motoko , Oohara Tadao , Makino Susumu , Wanibuchi Hideki , Fukushima Shoji
Published: August 2005

We previously reported p53 mutations to be frequent (greater than 70%), whereas both H‐ras mutations and microsatellite instability (MSI) were infrequent (about 10%), in urinary bladder carcinomas (UBCs) and their metastatic foci in the N‐butyl‐N‐(4‐hydroxybutyl)nitrosamine (BBN)‐induced mouse urothelial carcinogenesis model. In the present study, an analysis of p53 and H‐ras mutations as well as MSI was performed on 12 renal pelvic carcinomas (RPCs) and 8 metastatic or invading foci produced by the same experimental procedure. Histologically, 10 of the RPCs were transitional cell carcinomas and the remaining 2 were squamous cell carcinomas. p53 mutations were infrequent and only found in one primary RFC (8%), its metastatic foci and an invading lesion in another animal (in a total 2 of 12; 17%). H‐ras mutations were slightly more frequent (found in 3 of 12 animals; 25%), 4 of 5 involving codon 44, GTG to GCG, not a hot‐spot reported for human cancers. In two cases, H‐ras mutations were confined to lung metastasis and not detectable in their primary RPCs. MSI analysis was available for 6 pairs of primary RPCs and their metastatic foci, and 4 animals (67%) had MSI at one or more microsatellite loci. Overall, the distribution of genetic alterations differed from that in UBCs produced by the same experimental protocol. The results thus suggest that different genetic pathways may participate in carcinogenesis of the upper and lower urinary tract due to BBN.
A Statistical Study of Lung Cancer in the Annual of Pathological Autopsy Cases in Japan, from 1958 to 1997, with Reference to Time Trends of Lung Cancer in the World
Authors: Morita Toyohiko
Published: August 2005

Lung cancer cases (66 650 males and 20 890 females) registered in the Annual of Pathological Autopsy Cases in Japan, between 1958 and 1997, were analyzed with regard to sex, age and histology. They were subdivided into decades (periods I to IV), and compared with the Japanese mortality statistics, with which they were in good correspondence. Although the autopsy rate is decreasing, more than 10% of the total lung cancer deaths in Japan were registered by 1990. Among autopsied cases, the incidence of lung cancer cases increased from 6% to 12% in males and from 3% to 6% in females. From period III, lung cancer in males became the most frequent, and was the second most frequent cancer in females after gastric cancer. As for the histological distribution, adenocarcinoma was the most frequent and squamous cell carcinoma was the next most frequent in both sexes. Recently, a significant increase in adenocarcinoma and a significant decrease in squamous cell carcinoma have been observed in both sexes. The peak ages shifted from the 60s to the 70s and a significant rise in the mean ages were observed. The male‐to‐female ratios and the ratio curves by histological and age group were high for squamous cell carcinoma and small cell carcinoma, while they were low for the total and adenocarcinoma. With the recently acquired data on lung cancer in Asian countries, and from the male‐to‐female ratios and the status of smoking rates, lung cancer in the world at present was divided into three groups; North America, Europe and Asia. The possibility of one group changing to resemble another and of groups converging in the near future is suggested.
Identification of the CAB2/hCOS16 Gene Required for the Repair of DNA Double‐strand Breaks on a Core Amplified Region of the 17ql2 Locus in Breast and Gastric Cancers
Authors: Nezu Masahiko , Nishigaki Michiko , Ishizuka Tomoki , Kuwahara Yoshitaka , Tanabe Chikako , Aoyagi Kazuhiko , Sakamoto Hiromi , Saito Yasushi , Yoshida Terahiko , Sasaki Hiroki , Terada Masaaki
Published: August 2005

We previously reported that CAB1 and c‐ERBB‐2 genes were found to be located in a core amplified region of the 17q12 locus, which is frequently amplified in various cancers. During identification of this core region, CAB2, a human homologue of the yeast COS16 required for the repair of DNA double‐strand breaks was cloned. Autofluorescence analysis of cells transfected with its GFP fusion protein demonstrated that CAB2 translocates into vesicles, suggesting that overexpression of CAB2 may decrease intercellular Mn2+ by accumulating it in the vesicles, in the same way as yeast COS16. This is the first report identifying all of the genes on the core amplified region of the 17q12 locus in breast and gastric cancers.
Analysis of the β‐Catenin/T Cell Factor Signaling Pathway in 36 Gastrointestinal and Liver Cancer Cells
Authors: Ikenoue Tsuneo , Ijichi Hideaki , Kato Naoya , Kanai Fumihiko , Masaki Tsutomu , Rengifo William , Okamoto Makoto , Matsumura Masayuki , Kawabe Takao , Shiratori Yasushi , Omata Masao
Published: August 2005

We investigated the frequency and mechanism of |bT‐catenin/T cell factor (Tcf) signaling activation in a panel of 36 human gastrointestinal and liver cancer cell lines. Reporter assay and electrophoretic mobility shift assay revealed that the β‐catenin/Tcf signaling was upregulated in 12 of 12 (100%) colorectal, 5 of 8 (68%) gastric, 2 of 7 (29%) hepatic, and none of 9 pancreatic cancer cell lines. The activation of the pathway was mainly due to the mutation of adenomatous polyposis coli (APC) or β‐catenin, and Tcf‐4 was highly expressed in these cell lines with upregulated signaling. Nuclear β‐catenin was observed not only in the signaling‐activated cell lines, but also in 14 of 25 (56%) primary gastric cancers, 15 of 20 (75%) colon cancers, 5 of 19 (26%) hepatocellular carcinomas, and none of 13 pancreatic cancers. The presence of signaling‐upregulated gastric cancer cell lines with intact APC and β‐catenin suggests the involvement of other mechanisms than mutations of APC or β‐catenin.
Incorporation of the Anticancer Agent KRN5500 into Polymeric Micelles Diminishes the Pulmonary Toxicity
Authors: Mizumura Yasuo , Matsumura Yasuhiro , Yokoyama Masayuki , Okano Teruo , Kawaguchi Takanori , Moriyasu Fuminori , Kakizoe Tadao
Published: August 2005

KRN5500 is a highly active new semi‐synthetic water‐insoluble anticancer agent. The only mechanism of anticancer activity of KRN5500 described so far is an inhibitory effect on protein synthesis. At the tune of writing, a phase I clinical trial is under way at the National Cancer Center Hospital, Tokyo, and at the National Cancer Institute in the USA. Although preclinical data did not indicate lung toxicity, some cases of severe pulmonary disorder were reported in the phase I clinical trials. This study has been conducted to examine whether incorporation of KRN5500 into polymeric micelles (KRN/m) could reduce the toxic effects caused by the current formulation of KRN5500. The in vitro and in vivo antitumor activities of KRN5500 and KRN/m were compared. Pulmonary toxicity of KRN5500 and KRN/m was studied using a bleomycin (BLM)‐induced lung injury rat model. In BLM‐rats, extensive pulmonary hemorrhage with diapedesis was observed with KRN5500 i.v. bolus injection at the dose of 3 mg/kg, which is equivalent to 21.0 mg/m2 (level 5) of the Japanese phase I trial. However, toxicity was not observed when rats were administered KRN/m at the equivalent dose to KRN5500 in potency. Electron microscopy of the lung treated with KRN5500 showed disruption of the alveolar type II membrane with release of lamellar debris. Furthermore, in vivo, KRN/m showed similar antitumor activity to KRN5500. These results indicate that KRN/m may be useful for reducing the pulmonary toxicity associated with the current formulation of KRN5500, while fully maintaining its antitumor activity.
Molecular Analysis of Oncogenes, ras Family Genes (N‐ras, K‐ras, H‐ras), myc Family Genes (c‐myc, N‐myc) and mdm2 in Natural Killer Cell Neoplasms
Authors: Sugimoto Kei‐ji , Kawamata Norihiko , Sakajiri Sakura , Oshimi Kazuo
Published: August 2005

Natural killer (NK) cell neoplasms are rare diseases. Frequent abnormalities of the tumor suppressor genes Rb, p53, p151NK4B, p161NK4A and p14ARF have been reported. However, no oncogenes associated with tumorigenesis of NK cell neoplasms have been reported so far. We analyzed the status of oncogenes including N‐ras, K‐ras, H‐ras, c‐myc, N‐myc and mdm2 by Southern blot, PCR‐SSCP, western blot analysis and immunohistochemical staining. We analyzed four cell lines derived from NK cell neoplasms and 31 clinical samples with five subclasses of NK cell neoplasms. We found no point mutations of the ras family genes. We detected no mutations in the c‐myc and N‐myc genes. No overexpression of c‐Myc protein was detected by western blot analysis. Although we found neither amplification nor rearrangement of the mdm2 gene, we found high expression of MDM2 protein in some cases by western blot analysis. Immunohistochemical staining confirmed the overexpression of MDM2 protein. We found 14 cases with overexpression of MDM2 protein out of 15 cases (93%) with four subclasses of NK cell neoplasms except chronic NK lymphocytosis. Our previous and these results suggested that the expression level of MDM2 protein is independent of the status of the p14ARF, p53, Rb genes. MDM2 protein might independently contribute to car‐cinogenesis of NK cell neoplasms. Although the number of the cases we analyzed was not large, alterations of ras and myc family genes may rarely contribute to tumorigenesis in NK cell neoplasms. In contrast, overexpression of MDM2 might be associated with tumorigenesis of NK cell neoplasms, especially aggressive subclasses.
Serine Proteinase Inhibitor 9 Can Be Recognized by Cytotoxic T Lymphocytes of Epithelial Cancer Patients
Authors: Tanaka Koji , Harashima Nanae , Niiya Fumihiko , Miyagi Yoshiaki , Hida Naoya , Ochi Mika , Imai Nobue , Harada Mamoru , Itoh Kyogo , Shichijo Shigeki
Published: August 2005

Serine proteinase inhibitor 9 (PI–9) inhibits granzyme B‐mediated apoptosis and interleukin–lβ‐converting enzyme activity. In this study, we report that the PI–9 gene encodes antigenic epitopes recognized by the HLA‐A24–restricted and tumor‐reactive cytotoxic T lymphocytes (CTLs) of epithelial cancer patients. Screening of an autologous cDNA library using a CTL line recognizing HLA‐A24+ tumor cells resulted in the isolation of a cDNA, which had an identical coding region to the previously described PI–9 genes. PI–9 gene was expressed in approximately three‐fourths of epithelial cancer cell lines and all leukemic cell lines tested. It was also expressed in normal peripheral blood mononuclear cells (PBMCs), but not in a normal fibroblast cell line. CTL sublines contained T cells capable of recognizing the PI–9292–300 and PI–9348–356 peptides among 13 different peptides having the HLA‐A24 binding motifs. These two peptides were recognized by the CTL line in a dose‐dependent and HLA class‐I‐restricted manner, and also possessed the ability to induce HLA class I‐restricted and tumor‐reactive CTLs in PBMCs from HLA‐A24+ cancer patients. These results demonstrate that PI–9 is recognized by HLA class I‐restricted and tumor‐reactive CTLs of epithelial cancer patients.
Expression of PPARγ and Its Ligand‐dependent Growth Inhibition in Human Brain Tumor Cell Lines
Authors: Kato Mihoko , Nagaya Takashi , Fujieda Miyuki , Saito Kiyoshi , Yoshida Jun , Seo Hisao
Published: August 2005

Peroxisome proliferator‐activated receptor y (PPARγ) belongs to a superfamily of thyroid/steroid hormone receptors and regulates transcription of their target genes in a ligand‐dependent manner. Recently, PPARγ was reported to be expressed in several cell lines derived from breast, colon, stomach and lung cancers. Activation of PPARγ by its ligand inhibits the growth of these tumor cells, suggesting that PPARγ ligand is a potential anti‐cancer agent in PPARγ‐expressing tumors. However, its expression in brain tumors has not been studied. We thus studied the expression in glioma samples with different pathological stages from 20 patients. It was demonstrated that 95% of the glioma tissue expressed PPARγ mRNA. The results prompted us to study whether PPARγ ligand affects the growth of cell lines derived from brain tumors. The receptor expression was studied in 9 cell lines either derived from malignant glioma or neuroblastoma. The expression was detected in a glioma cell line SK‐MG‐1 and in a neuroblastoma cell line NB‐1. Addition of one of the PPARγ ligands, troglitazone, induced growth inhibition in both cell lines. Further analyses revealed that this growth inhibition is caused by a PPARγ‐mediated induction of apoptosis. These results suggest that PPARγ ligands could be a potential therapeutic agent for the treatment of the brain tumors expressing this receptor.
Cell‐free Human T‐cell Leukemia Virus Type 1 Binds to, and Efficiently Enters Mouse Cells
Authors: Sun Binlian , Nitta Takayuki , Shoda Momoko , Tanaka Masakazu , Hanai Shuji , Hoshino Hiroo , Miwa Masanao
Published: August 2005

Human T‐cell leukemia virus type 1 (HTLV–1) is an etiologic agent of adult T‐cell leukemia/lymphoma and other HTLV‐1–associated diseases. However, the interaction between HTLV–1 and T cells in the pathogenesis of these diseases is poorly understood. Mouse cells have been reported to be resistant to cell‐free HTLV–1 infection. However, we recently reported that HTLV–1 DNA could be observed 24 h after cell‐free HTLV–1 infection of mouse cell lines. To understand HTLV–1 replication in these cells in detail, we concentrated the virus produced from c77 feline kidney cell line and established an efficient infection system. The amounts of adsorption of HTLV–1 are larger in mouse T cell lines, EL4 and RLml, than those in human T cell lines, Molt4 and HUT78, and are similar to that in human kidney cell line, 293T. Unexpectedly, however, the amounts of entry of HTLV–1 are about 10–fold larger in the two mouse cell lines than those in the three human cell lines employed. Moreover, viral DNA was detectable from 1 h in EL4 and RLml cells, but only from 2–3 h in 293T, Molt4 and HUT78 cells. However, the amount of viral DNA in EL4 cells became smaller than that in Molt4 cells. HTLV–1 expression could be detected until day 1–2 in RLml and EL4 cells, and until day 4 in Molt4 cells. Our results suggest that mouse cell experiments would give useful information to dissect the early steps of cell‐free HTLV–1 infection.
Downregulation of the nuclear‐encoded subunits of the complexes III and IV disrupts their respective complexes but not complex I in procyclic Trypanosoma brucei
Authors: Horváth Anton , Horáková Eva , Dunajčíková Petra , Verner Zdeněk , Pravdová Eliška , Šlapetová Iveta , Cuninková L’udmila , Lukeš Julius
Published: August 2005

The function, stability and mutual interactions of selected nuclear‐encoded subunits of respiratory complexes III and IV were studied in the Trypanosoma brucei procyclics using RNA interference (RNAi). The growth rates and oxygen consumption of clonal cell lines of knock‐downs for apocytochrome c1 (apoc1) and the Rieske Fe‐S protein (Rieske) of complex III, and cytochrome c oxidase subunit 6 (cox6) of complex IV were markedly decreased after RNAi induction. Western analysis of mitochondrial lysates using specific antibodies confirmed complete elimination of the targeted proteins 4–6 days after induction. The Rieske protein was reduced in the apoc1 knock‐down and vice versa, indicating a mutual interdependence of these components of complex III. However, another subunit of complex IV remained at the wild‐type level in the cox6 knock‐down. As revealed by two‐dimensional blue native/SDS‐PAGE electrophoresis, silencing of a single subunit resulted in the disruption of the respective complex, while the other complex remained unaffected. Membrane potential was reproducibly decreased in the knock‐downs and the activities of complex III and/or IV, but not complex I, were drastically reduced, as measured by activity assays and histochemical staining. Using specific inhibitors, we have shown that in procyclics with depleted subunits of the respiratory complexes the flow of electrons was partially re‐directed to the alternative oxidase. The apparent absence in T. brucei procyclics of a supercomplex composed of complexes I and III may represent an ancestral state of the respiratory chain.
Novel lipopolysaccharide biosynthetic genes containing tetranucleotide repeats in Haemophilus influenzae, identification of a gene for adding O‐acetyl groups
Authors: Fox Kate L. , Yildirim Håkan H. , Deadman Mary E. , Schweda Elke K. H. , Moxon E. Richard , Hood Derek W.
Published: August 2005

Many of the genes for lipopolysaccharide (LPS) biosynthesis in Haemophilus influenzae are phase variable. The mechanism of this variable expression involves slippage of tetranucleotide repeats located within the reading frame of these genes. Based on this, we hypothesized that tetranucleotide repeat sequences might be used to identify as yet unrecognized LPS biosynthetic genes. Synthetic oligonucleotides (20 bases), representing all previously reported LPS‐related tetranucleotide repeat sequences in H. influenzae, were used to probe a collection of 25 genetically and epidemiologically diverse strains of non‐typeable H. influenzae. A novel gene identified through this strategy was a homologue of oafA, a putative O‐antigen LPS acetylase of Salmonella typhimurium, that was present in all 25 non‐typeable H. influenzae, 19 of which contained multiple copies of the tetranucleotide 5′‐GCAA. Using lacZ fusions, we showed that these tetranucleotide repeats could mediate phase variation of this gene. Structural analysis of LPS showed that a major site of acetylation was the distal heptose (HepIII) of the LPS inner‐core. An oafA deletion mutant showed absence of O‐acetylation of HepIII. When compared with wild type, oafA mutants displayed increased susceptibility to complement‐mediated killing by human serum, evidence that O‐acetylation of LPS facilitates resistance to host immune clearance mechanisms. These results provide genetic and structural evidence that H. influenzae oafA is required for phase variable O‐acetylation of LPS and functional evidence to support the role of O‐acetylation of LPS in pathogenesis.
Characterization of the 26S rRNA‐binding domain in Saccharomyces cerevisiae ribosomal stalk phosphoprotein P0
Authors: Santos Cruz , Ballesta Juan P. G.
Published: August 2005

The stalk is a universal structure of the large ribosomal subunit involved in the function of translation factors. The bacterial stalk is highly stable but its stability is notably reduced in eukaryotes, favouring a translation regulatory activity of this ribosomal domain, which has not been reported in prokaryotes. The RNA‐binding protein P0 plays a key role in determining the eukaryotic stalk activities, and characterization of the P0/RNA interaction is essential to understand the evolutionary process. Using a series of Saccharomyces cerevisiae‐truncated proteins, a direct involvement of two N‐terminal regions, I3‐M58 and K81‐V121, in the interaction of P0 with the ribosome has been shown. Two other conserved regions, R122‐T149 and G162‐T182, affect P0 interaction with other stalk components and the sensitivity to sordarin anti‐fungals but are not essential for RNA binding. Moreover, P0 and a P0 fragment comprising only the first 121 amino acids show a similar in vitro affinity for the highly conserved 26S rRNA binding site. A protein chimera containing the first 165 amino acids of L10, the P0 bacterial counterpart, is able to complement the absence of P0 and also shows the same P0 RNA binding characteristics. Altogether, the results indicate that the affinity of the stalk RNA‐binding protein for its substrate has been highly conserved, and changes in the stability of the interaction of P0 with the ribosome, which are essential for the new eukaryotic functions, result from the evolution of the overall stalk structure.
Zebra dolomitization as a result of focused fluid flow in the Rocky Mountains Fold and Thrust Belt, Canada
Authors: VANDEGINSTE VEERLE , SWENNEN RUDY , GLEESON SARAH A. , ELLAM ROB M. , OSADETZ KIRK , ROURE FRANÇOIS
Published: August 2005

Discordant zebra dolomite bodies occur locally in the Middle Cambrian Cathedral and Eldon Formations of the Main Ranges of the Canadian Rocky Mountains Fold and Thrust Belt. They are characterized by alternating dark grey (a) and white (b) bands, forming an ‘abba’ diagenetic cyclicity. These bands developed parallel to both bedding and cleavage. Dark grey (a) bands consist of fine (< 300 μm) non‐planar crystalline impure dolomite. The white (b) bands are composed of coarse (up to several millimetres) milky‐white pure saddle dolomites (b1) which are often covered by pore‐lining zoned dolomite (b2). Thebphases often possess a saddle‐shaped morphology. In contrast to the replacement origin of theadolomite, the zonedb2dolomite rims are interpreted as a cement formed in open cavities. Theb1dolomite is interpreted as the result of recrystallization with diagenetic leaching of non‐carbonate components.

All the zebra dolomites studied are (nearly) stoichiometric and are characterized by enriched Na and depleted Sr concentrations. Fe and Mn concentrations in these dolomites differ depending on the sample locality. Fluid inclusion data indicate that the dolomites formed from relatively hot (TH = 130–200 °C), saline (20–23 wt% CaCl2eq.) fluids. A diagenetic high temperature origin is also supported by depletedδ18O values (−20 to −14‰ VPDB). A contribution of87Sr‐enriched fluids is reflected in the87Sr/86Sr values (0·7091–0·7123).

Zebra dolomite development is explained by focused fluid flow, which exploited areas of structural weaknesses (e.g. basin‐platform, rim areas, faults, etc.). Expulsion of hot basinal brines in a tectonically active regime generated overpressures, which explains the development of secondary porosity during zebra dolomitization as well as the intra‐zebra fracturing at decimetre to micrometre scale.
A genetic map ofCottus gobio(Pisces, Teleostei) based on microsatellites can be linked to the physical map ofTetraodon nigroviridis
Authors: STEMSHORN K. C. , NOLTE A. W. , TAUTZ D.
Published: August 2005

To initiate QTL studies in the nonmodel fishCottus gobiowe constructed a genetic map based on 171 microsatellite markers. The mapping panel consisted ofF1intercrosses between two divergentCottuslineages from the River Rhine System. Basic local alignment search tool (BLAST) searches with the flanking sequences of the microsatellite markers yielded a significant (e < 10−5) hit with theTetraodon nigroviridisgenomic sequence for 45% of theCottusloci. Remarkably, most of these hits were due to short highly conserved noncoding stretches. These have an average length of 40 bp and are on average 92% conserved. Comparison of the map locations between the two genomes revealed extensive conserved synteny, suggesting that theTetraodongenomic sequence will serve as an excellent genomic reference for at least the Acanthopterygii, which include evolutionarily interesting fish groups such as guppies (Poecilia), cichlids (Tilapia) orXiphophorus(Platy). The apparent high density of short conserved noncoding stretches in these fish genomes will highly facilitate the identification of genes that have been identified in QTL mapping strategies of evolutionary relevant traits.
Hybridization dynamics between sympatric species of trout: loss of reproductive isolation
Authors: BETTLES C. M. , DOCKER M. F. , DUFOUR B. , HEATH D. D.
Published: August 2005

Although reinforcement should enhance reproductive barriers in sympatric species, sympatric trout species do hybridize. Using mitochondrial and nuclear species markers, we investigated hybridization directionality, hybrid mating biases, and selection against hybrids in 13 sympatric cutthroat and rainbow trout populations on Vancouver Island, Canada. Approximately 50% of the genotyped fish were hybrid (F1 or higher‐order) and populations ranged from very recent (all F1 hybrids) to extremely advanced higher‐order hybridization. Overall, interbreeding was reciprocal, although some populations showed directional hybridization. Pronounced cytonuclear disequilibrium in post‐F1 hybrids indicated a remarkable mating bias not previously reported, which is most likely because of behavioural reproductive preferences. Selection against hybrids was observed in only two populations, indicative of extrinsic selection. Two populations were ‘hybrid swarms’, with a complete loss of reproductive isolation. The complex hybridization dynamics in this system represent a valuable natural experiment of the genetic and evolutionary implications of recent and on‐going interspecific hybridization.
Pollinating fig waSPS: genetic consequences of island recolonization
Authors: ZAVODNA MONIKA , ARENS PAUL , VAN DIJK PETER J. , PARTOMIHARDJO TUKIRIN , VOSMAN BEN , VAN DAMME JOS M. M.
Published: August 2005

The levels of genetic diversity and gene flow may influence the long‐term persistence of populations. Using microsatellite markers, we investigated genetic diversity and genetic differentiation in island (Krakatau archipelago, Indonesia) and mainland (Java and Sumatra, Indonesia) populations of Liporrhopalum tentacularis and Ceratosolen bisulcatus, the fig wasp pollinators of two dioecious Ficus (fig tree) species. Genetic diversity in Krakatau archipelago populations was similar to that found on the mainland. Population differentiation between mainland coastal sites and the Krakatau islands was weak in both wasp species, indicating that the intervening 40 km across open sea may not be a barrier for wasp gene flow (dispersal) and colonization of the islands. Surprisingly, mainland populations of the fig waSPS may be more genetically isolated than the islands, as gene flow between populations on the Javan mainland differed between the two wasp species. Contrasting growth forms and relative ‘immunity’ to the effects of deforestation in their host fig trees may account for these differences.
Sexual conflict and female immune suppression in the cricket, Allonemobious socius
Authors: FEDORKA KENNETH M. , ZUK MARLENE
Published: August 2005

In many animal systems, females exhibit a localized immune response to insemination that helps defend against sexually transmitted disease. However, this response may also kill sperm, reducing a male's reproductive potential. If males could suppress this response, they may be able to increase their sperm's representation in the female's reproductive tract, thereby increasing their fitness. Here we address the hypothesis that, under conditions of sperm competition, males interfere with female immunity. To test our hypothesis, we manipulated levels of female mating frequency (single vs. multiply mated) and seminal diversity (monandrous vs. polyandrous) in the cricket, Allonemobius socius and measured female immune response. As mating frequency increased, female hemocyte load decreased, indicating a general reproductive cost. As seminal diversity increased, phenoloxidase (PO) activity (in vitro measure of ‘potential’ macroparasitic defense) increased and encapsulation ability (in vivo measure of ‘realized’ macroparasitic defense) decreased in polyandrous females. These results suggest that males may manipulate female immunity by interrupting the pro‐PO cascade, which begins with the activation of PO and ends in the encapsulation of invading foreign bodies. In other words, female immune function may serve as a battleground over which a sexual conflict is fought.
Asymmetrical reproductive character displacement in the house mouse
Authors: SMADJA C. , GANEM G.
Published: August 2005

Our study addressed reproductive character displacement between two subspecies of the house mouse,Mus musculus musculusandMus musculus domesticusthat hybridize in Europe along a zone where selection against hybridization is known to occur. Based on a multi‐population approach, we investigated spatial patterns of divergence of mate preference in the two taxa. Mate preference was significantly higher in the contact zone than in allopatry in both subspecies, suggesting that reproductive character displacement occurs. Moreover, patterns of preference were stronger inM. m. musculusthan inM. m. domesticus, indicating an asymmetrical divergence between the two. In the context of selection against hybridization, our results may provide empirical support for the hypothesis of reinforcement in a parapatric hybrid zone. We discuss factors that could explain the asymmetrical pattern of divergence and the possible impact of a unimodal structure on the maintenance of premating divergence between the two subspecies.
Life‐history invariants with bounded variables cannot be distinguish from data generated by random processes using standard analyses
Authors: CIPRIANI R. , COLLIN R.
Published: August 2005

A dimensionless approach to the study of life‐history evolution has been applied to a wide variety of variables in the search for life‐history invariants. This approach usually employs ordinary least squares (OLS) regressions of log‐transformed data. In several well‐studied combinations of variables the range of values of one parameter is bounded or limited by the value of the other. In this situation, the null hypothesis normally applied to regression analysis is not appropriate. We generate the null expectations and confidence intervals (CI) for OLS and reduced major axis (RMA) regressions using random variables that are bounded in this way. Comparisons of these CI show that, for log‐transformed data, the patterns generated by random data and those predicted by life history invariant theory often could not be distinguished because both predict a slope of 1. We recommend that tests based on the putative invariant ratios and not the correlations between the two variables be used in the exploration of life‐history invariants using bounded data. Because empirical data are often not normally distributed randomization test may be more appropriate than standard statistical tests.
Indirect consequences of artificial selection on plasticity to light quality inArabidopsis thaliana
Authors: CALLAHAN H. S. , PIGLIUCCI M.
Published: August 2005

Covariation between light quality‐ and photoperiod‐mediated phenotypic plasticity was investigated usingArabidopsis thaliana.Three episodes of artificial selection were imposed on an index that quantified the plastic response to reduced red to far‐red ratios (R : FR), with higher index values indicating greater plasticity. Relative to control lines, two high plasticity (HP) lines showed 1.6‐ and 2.4‐fold increases in the index; low plasticity (LP) lines showed 1.4‐ and 1.1‐fold decreases. A factorial experiment combining high and low R : FR conditions with long and short photoperiods assessed indirect consequences of selection on plasticity. Despite divergent R : FR‐mediated plasticities in HP vs. LP lines, all four lines showed increases in photoperiod‐mediated responses and decreases in mean leaf number. Complex relationships among trait means, plasticities and underlying mechanisms caution against generalizing about the genetic architecture of plastic traits. Partially independent developmental and evolutionary responses to R : FR and photoperiod are somewhat unsurprising, given this species’ cosmopolitan nature.
The mode of evolution of aggregation pheromones in Drosophila species
Authors: SYMONDS M. R. E. , WERTHEIM B.
Published: August 2005

Aggregation pheromones are used by fruit flies of the genus Drosophila to assemble on breeding substrates, where they feed, mate and oviposit communally. These pheromones consist of species‐specific blends of chemicals. Here, using a phylogenetic framework, we examine how differences among species in these pheromone blends have evolved. Theoretical predictions, genetic evidence, and previous empirical analysis of bark beetle species, suggest that aggregation pheromones do not evolve gradually, but via major, saltational shifts in chemical composition. Using pheromone data for 28 species of Drosophila we show that, unlike with bark beetles, the distribution of chemical components among species is highly congruent with their phylogeny, with closely related species being more similar in their pheromone blends than are distantly related species. This pattern is also strong within the melanogaster species group, but less so within the virilis species group. Our analysis strongly suggests that the aggregation pheromones of Drosophila exhibit a gradual, not saltational, mode of evolution. We propose that these findings reflect the function of the pheromones in the ecology of Drosophila, which does not hinge on species specificity of aggregation pheromones as signals.