bluuebunny/crossref_metadata_2025 huggingface dataset:
Survival Rates of Childhood Cancer Patients in Osaka, Japan, 1975–1984
Authors: Ajiki Wakiko , Hanai Aya , Tsukuma Hideaki , Hiyama Tomohiko , Fujimoto Isaburo
Published: August 2005

Survival rates for childhood cancers were analyzed with a total of 2,209 cases who were registered in a population‐based cancer registry in Osaka, Japan in 1975–1984. These cases were reclassified according to Birch's classification and the survival rate of each diagnostic group was calculated by Kaplan‐Meier methods. Death certificate‐only cases, which amounted to 3.9% of all incidence, were excluded from the calculation. The five‐year cumulative survival rate for both sexes was 46% for all cancer children. Among 12 major diagnostic groups, the most favorable survival was seen in retinoblastoma (87.5%), followed by renal tumors, epithelial neoplasms, and gonadal and germ‐cell tumors. The outcome was unfavorable in leukemias, sympathetic nervous system tumors, hepatic tumors and malignant bone tumors. Comparing the survival in 1975–1979 with that in 1980–1984, the rate for all childhood cancer rose from 41% to 51%. Improvement in survival was also observed in 4 groups; acute lymphocytic leukemia, acute non‐lymphocytic leukemia, non‐Hodgkin's lymphoma and osteosarcoma. One attributable factor for the rise of survival was proved to be improvement of medical treatment by Cox's hazard model analysis. Comparison of survival rates in Osaka with those in England and the U.S. revealed that the prognosis for acute lymphocytic leukemia and acute non‐lymphocytic leukemia was less favorable in Osaka than in England and the U.S.
Protective Effects of Kampo Medicines and Baicalin against Intestinal Toxicity of a New Anticancer Camptothecin Derivative, Irinotecan Hydrochloride (CPT‐11), in Rats
Authors: Takasuna Kiyoshi , Kasai Yoshio , Kitano Yutaka , Mori Kazuhiko , Kobayashi Reiko , Hagiwara Takehiro , Kakihata Kohji , Hirohashi Masaaki , Nomura Mamoru , Nagai Eiichi , Kamataki Tetsuya
Published: August 2005

In clinical use, irinotecan hydrochloride (CPT‐11; 7‐ethyl‐10‐[4‐(piperidmo)‐l‐piperidino]carbonyl‐oxycamptothecin), a novel antitumor agent, causes a relatively high incidence of severe forms of diarrhea. We investigated whether baicalin, an inhibitor of β‐glucuronidase, which deconjugates the glucuronide of the active metabolite of CPT‐11, SN‐38 (7‐ethyl‐10‐hydorxycamptothecin), and Japanese herbal medicines (Kampo medicines) which contain baicalin can ameliorate CPT‐11‐induced intestinal toxicity in rats. CPT‐11 (60 mg/kg i.v. once daily for 4 consecutive days) induced intestinal toxicity characterized by diarrhea, loss of body weight, anorexia and disruption of intestinal epithelium. Treatment with baicalin (25 mg/kg p.o. twice daily) or Kampo medicines (TJ‐14 and TJ‐114; 1 g/kg p.o. twice daily) from the day before to 4 or 10 days after the start of CPT‐11 administration resulted in significantly decreased weight loss, improved anorexia and a delayed onset of diarrheal symptoms. Histological examination revealed that Kampo medicine‐treated animals had less damage to the intestinal epithelium and that damage was repaired more rapidly than in control rats. These results suggest that the prophylactic use of Kampo medicines (TJ‐14 and TJ‐114) may be of value against CPT‐11‐induced intestinal toxicity.
Basic Fibroblast Growth Factor as a Candidate Tumor Marker for Renal Cell Carcinoma
Authors: Fujimoto Kiyohide , Ichimori Yuzo , Yamaguchi Hisako , Arai Kunihiko , Futami Takashi , Ozono Seiichiro , Hirao Yoshihiko , Kakizoe Tadao , Terada Masaaki , Okajima Eigoro
Published: August 2005

The present investigation was conducted to determine serum levels of basic fibroblast growth factor (FGF) by enzyme immunoassay in patients with various urogenital tumors. Renal cell carcinoma had a higher tendency (28 of 52, 53.8%) toward increased serum levels of basic FGF than any of the other urogenital tumors, and increased serum basic FGF was detected more frequently in patients with advanced renal cell carcinoma. Analysis of histological pattern indicated that renal cell carcinoma with a solid or tubular component is more likely to produce basic FGF. However, no significant difference was seen between the percentage of clear cell type tumor patients with increased serum basic FGF (50.0%) and the percentage of granular cell type tumor patients with increased serum basic FGF (66.7%). Five of 8 patients with renal cell carcinoma who underwent selective renal venous sampling before nephrectomy showed increased serum basic FGF in the renal vein from the affected kidney. After resection of the affected kidney to remove the cancer, serum basic FGF disappeared within 2 weeks. However, residual huge tumor or postoperative disease prolonged the increased levels of basic FGF in 2 patients, indicating that basic FGF is produced from and secreted by tumor tissue itself. These findings suggest that serum basic FGF can be useful in the diagnosis, and in evaluating the prognosis, of patients with renal cell carcinoma.
Correlation of Gene‐specific Damage with Cisplatin between Human Adenocarcinoma Cells and Peripheral Blood Mononuclear Cells Analyzed by Polymerase Chain Reaction‐stop Assay
Authors: Oshita Fumihiro , Arioka Hitoshi , Heike Yuji , Shiraishi Jun‐ichi , Saijo Nagahiro
Published: August 2005

We investigated gene‐specific damage in adenocarcinoma cells, obtained from pleural effusions of 9 primary lung cancer patients, induced by incubation with cisplation for 3 h in vitro. The 2.7 kb fragment of the hypoxanthine phosphoribosyltransferase (HPRT) gene was amplified by the polymerase chain reaction (PCR) to quantify the DNA damage. A 7‐fold difference in the extent of gene‐specific damage among the patients was observed. Mononuclear cells (MNC) were obtained from freshly isolated blood from the same patients before they received chemotherapy. These cells were also incubated with cisplatin in vitro, and PCR amplification of the HPRT gene was carried out. A 4‐fold variation of DNA damage among the patients was observed. Moreover, there was a linear correlation between the extents of the DNA damage in the tumor cells and MNCs (R2=0.676, P=0.0016). These results suggest that the PCR‐stop assay could be used to detect interindividual variations in the extent of gene‐specific damage in both tumor cells and MNC from the same patients induced by cisplatin treatment. In conclusion, MNC could be used to analyze cisplatin‐induced gene‐specific damage in cancer patients whose tumor cells are inaccessible.
An Rsa I Polymorphism in the CYP2E1 Gene Does Not Affect Lung Cancer Risk in a Japanese Population
Authors: Watanabe Junko , Yang Jian‐Ping , Eguchi Hidetaka , Hayashi Shin‐ichi , Imai Kazue , Nakachi Kei , Kawajiri Kaname
Published: August 2005

CYP2E1 catalyzes the metabolic activation of tobacco‐specific N‐nitrosamines, including 4‐(methylnitrosamino)‐1‐(3‐pyridyl)‐1‐butanone. An Rsa I polymorphism, which is located in the 5′‐flanking region of the CYP2E1 gene, has been found to affect the transcriptional regulation of the gene, resulting in different expression levels of the mRNA among individuals. In order to investigate an association between the Rsa I polymorphism and lung cancer susceptibility, the genotype distribution among 316 lung cancer patients was compared with that in 503 healthy controls. No statistically significant association was found between the Rsa I polymorphism and an increased risk of lung cancer, even though histological types of lung cancer, cigarette smoking and alcohol consumption were taken into account.
Presence of Streptococcus DNA Sequence in Surgical Specimens of Gastric Cancer
Authors: Sasaki Hiroki , Igaki Hiroyasu , Ishizuka Tomoki , Kogoma Yukiko , Sugimura Takashi , Terada Masaaki
Published: August 2005

In Southern blot analysis using Mycoplasma16S ribosomal DNA (rDNA) as a probe, positive signals were detected in DNA samples from surgical specimens of gastric cancers. The DNA that hybridized to Mycoplasma 16S rDNA was eluted from the gel, cloned and sequenced. The cloned sequence was identical to 16S rDNA of Streptococcus anginosus. In Southern blot analysis with the S. anginosus 16S rDNA fragment as a new probe, positive signals were detected in 9 (20%) out of 43 cases of gastric cancer.
Similarities of in situ mRNA Expression between Gelatinase A (MMP‐2) and Type I Procollagen in Human Gastrointestinal Carcinoma: Comparison with Granulation Tissue Reaction
Authors: Ohtani Haruo , Nagai Tadashi , Nagura Hiroshi
Published: August 2005

The mRNA localizations of gelatinase A (MMP‐2) and type I procollagen in human gastrointestinal carcinoma and non‐neoplastic fibrous granulation tissue were compared. By in situ hybridization, gelatinase A was shown to be expressed in fibroblasts not only in cancer stroma but also in the “reactive fibrosis zone,” which surrounds the invasive margin of cancer. In most cases, no accentuation of gelatinase A expression was observed in the invasive margin. This localization pattern of gelatinase A was essentially the same as that of type I procollagen. In gastric cancer associated with deep ulceration, gelatinase A was more abundantly expressed in fibroblastic cells in granulation tissue of the ulcer base than in cancer stroma. The same situation was found in non‐neoplastic fibrous granulation tissue, in which fibroblasts abundantly expressed mRNAs for both gelatinase A and type I collagen. Scar tissue (old fibrotic lesion) showed diminished expression of mRNAs for both gelatinase A and type I procollagen. The localization patterns suggest another function of gelatinase A in cancer tissue as a turnover enzyme of the extracellular matrix.
Inostamycin, an Inhibitor of P‐Glycoprotein Function, Interacts Specifically with Phosphatidylethanolamine
Authors: Kawada Manabu , Umezawa Kazuo
Published: August 2005

The mechanism of inostamycin action was further studied. When multidrug‐resistant KB‐C4 cells were preincubated with inostamycin for 30 min, the accumulation of [3H]vinblastine was increased for as long as 48 h thereafter. Inostamycin inhibited azidopine binding to P‐glycoprotein, even after KB plasma membranes had been preincubated with inostamycin and washed. Carbon 14‐labeled inostamycin bound to KB plasma membranes irreversibly, but the binding capacity did not parallel the amount of P‐glycoprotein in three KB cell lines. Inostamycin was found to interact specifically with purified phosphatidylethanolamine. These results suggest that inostamycin can inhibit P‐glycoprotein irreversibly by binding to plasma membranes irreversibly through phosphatidylethanolamine.
Hyperfractionated and Hypofractionated Radiation Therapy for Human Malignant Glioma Xenograft in Nude Mice
Authors: Hasegawa Masatoshi , Niibe Hideo , Mitsuhashi Norio , Yamakawa Michitaka , Kato Shingo , Furuta Masaya , Kazumoto Tomoko , Hayakawa Kazushige , Nakazato Yoichi
Published: August 2005

Xenografts of a human malignant glioma subcutaneously transplanted into nude mice were irradiated with graded single doses (2, 5, 10 or 20 Gy) or five types of fractionation schedules in two weeks: conventional [20 Gy in 10 fractions (fr)], hyperfractionated [24 Gy in 20 fr (two fractions per day)], and hypofractionated‐1, 2, 3 [20 Gy, 18 Gy, 16 Gy in 4 fr]. All of the fractionated irradiation groups showed tumor regression. The hypofractionation‐1 group (20 Gy in 4 fr) demonstrated the most prominent tumor regression, while the hyperfractionation group (24 Gy in 20 fr) showed the least effect. The hypofractionation‐2 group (18 Gy in 4 fr) showed similar regression to the conventional fractionation group (20 Gy in 10 fr). Histologically, tumors in the control groups consisted of a homogenous population of small anaplastic cells, and only a small number of tumor cells were glial fibrillary acidic protein (GFAP)‐positive. Following irradiation, the population of small anaplastic cells decreased and the percentage of GFAP‐positive cells increased. Cellular pleomorphism became much more prominent after irradiation in all of the fractionated irradiation groups as compared with the graded single dose irradiation groups. In this study, hyperfractionation was not effective against human glioma xenografts compared with conventional fractionation and hypofractionation. This indicates that care is needed in applying hyperfractionation regimens to human malignant gliomas.
Paneth Cell‐rich Flat Adenoma of the Rectum: Report of a Case
Authors: Rubio Carlos A. , Kanter Lena , Björk Jan , Poppen Bertil , Bry Lynn
Published: August 2005

A patient having familiar adenomatosis polyposis and an ileo‐rectal anastomosis developed a flat mucosal lesion in the rectum. A punch biopsy revealed a villous adenoma with high‐grade dysplasia. The subsequent surgical specimen indicated that the flat villous adenoma was rich in Paneth cells. Special stains included lysozyme muramidase (to visualize Paneth cells), MIB1 proliferation monoclonal antibody and single and multilabel immunohistochemistry for Paneth cells. Other methods included transmission electron microscopy and quantification with an image quantifier (Program Optilab 2.1) of lysozyme‐stained Paneth cells. The subjective evaluation of hematoxylin‐cosin‐stained preparations demonstrated that the Paneth cells were mainly located in the lower half of the villi. Sections labeled with a specific stain (lysozyme muramidase) revealed more Paneth cells in the villi and electron microscopy showed even more in lysozyme‐negative areas. Obviously some migrating dysplastic Paneth cells had retained their characteristic granules on their way towards the tip of the villi. Quantitative studies indicated that the lysozyme muramidase‐positive material accounted for 41% of the adenomatous tissue. MIB1 revealed intense cell proliferation at the base of the adenoma and in the entire slopes of the villi. Despite the wide distribution of Paneth cells in intestinal metaplasia of the stomach, in the normal small intestine and in the large bowel with chronic inflammatory diseases, it is surprising that tumors arising in Paneth cells are extremely rare. The causes of the apparent natural resistance of Paneth cells to tumor development deserve to be investigated. This is the first case of Paneth cell‐rich flat adenoma of the rectum in the literature.
Genetic and Epigenetic Resistance of SL/Ni Mice to Lymphomas
Authors: Shisa Hayase , Yamada Yoshihiro , Kawarai Atsuko , Terada Naoki , Kawai Makoto , Matsushiro Hisanori , Hiai Hiroshi
Published: August 2005

The murine spontaneous B lymphoma is etiologically related to the expression of endogenous ecotropic marine leukemia virus (ETV). Although both SL/Kh and SL/Ni mouse strains show a high level of expression of ETV from early in life, the former is a pre‐B lymphoma‐prone strain and the latter is rather lymphoma‐resistant. In order to identify the host background difference related to the lymphomagenesis, we performed a genetic cross study between these two strains. In the reciprocal F1 generation, the length of the lymphoma latent period was slightly but significantly longer in (SL/Ni XSL/Kh)F1 than in (SL/Kh × SL/Ni)F1 (P<0.05). The incidence of overall lymphomas and that of acute pre‐B lymphomas was lower in (SL/Ni × SL/Kh)F1 than in (SL/Kh × SL/Ni)F1, although the difference was not statistically significant. These observations indicate that an epigenetic maternal resistance mechanism of SL/Ni mice plays a role in the lymphoma resistance. Furthermore, in the backcross combinations without maternal influence of SL/Ni, we observed a genetic mechanism of lymphoma resistance: an SL/Ni‐derived recessive lymphoma‐resistance gene mapped in the proximal segment of Chr. 4. We named this gene nir‐1 (SL/Ni‐lymphoma resistance‐1). Thus, we have demonstrated epigenetic and genetic mechanisms of lymphoma resistance of the SL/Ni mouse with the high expression of endogenous ETV.
Absence of a Mutation of the p21/WAF1 Gene in Human Lung and Pancreatic Cancers
Authors: Shimizu Tatsunori , Miwa Wataru , Nakamori Shoji , Ishikawa Osamu , Konishi Yoichi , Sekiya Takao
Published: August 2005

The expression of a negative regulator of the cell cycle, p21WAF1 protein, is trans‐activated hy wild‐type p53, but not by the mutant protein. Therefore, mutations of the p53 and WAF1 genes may be complementary. We examined DNAs from 70 human primary lung (63 of NSCLC and 7 of SCLC) and 24 pancreatic cancers (19 primary cancers and 5 cell lines) for mutations of the WAF1 gene. No mutations were detected in any samples examined, regardless of the mutational state of the p53 gene. The results suggested that aberrations of the coding sequence of the WAF1 gene are not associated with carcinogenesis in lung and pancreatic cancers.
Deletion Mapping of Chromosome 10 in Human Glioma
Authors: Sonoda Yukihiko , Murakami Yoshinori , Tominaga Teiji , Kayama Takamasa , Yoshimoto Takashi , Sekiya Takao
Published: August 2005

We analyzed DNAs from 35 gliomas (27 malignant, grades III and IV; 8 less malignant, grades I and II) for loss of heterozygosity (LOH) using microsatellite sequences on chromosome 10 as polymorphic markers. An LOH was found in 8 of 11 (73%) glioblastomas (grade IV) and 4 of 16 (25%) grade III gliomas, but not in the less malignant types. We detected three commonly deleted regions. One was located in a telomeric region of 10p and the others were relatively large regions of 10q. Our results suggested that three putative tumor suppressor genes on chromosome 10 are involved in the malignant progression of gliomas.
Inhibition of Cell Growth by Transforming Growth Factor β1 Is Associated with p53‐Independent Induction of p21 in Gastric Carcinoma Cells
Authors: Akagi Morihisa , Yasui Wataru , Akama Yoshihiko , Yokozaki Hiroshi , Tahara Hidetoshi , Haruma Ken , Kajiyama Goro , Tahara Eiichi
Published: August 2005

Cell cycle regulators such as cyclins, cyclin‐dependent kinases (cdks) and their inhibitors control the growth of cells. SDI1/CIP1/WAF1/p21 is a potent inhibitor of G1 cdks, whose expression is induced by wild‐type p53. To elucidate the mechanism of growth inhibition by transforming growth factor β1 (TGFβ1), we examined the effect of TGPβ1 on the expression of p21, G1 cyclins and cdks by human gastric cancer cell lines. TGFβ1 induced p21 expression and subsequently suppressed cdk2 kinase activity, followed by a reduction in phosphorylation of the product of the retinoblastoma tumor suppressor gene in TMK‐1 cells, which are responsive to TGFβ1. Coimmunoprecipitation analysis demonstrated that TGFβ1 increased the level of p21 protein present in complexes with cdk2. In contrast, TGFβ1 did not induce p21 in TGFβ1‐resistant MKN‐28 cells. TGFβ1 did not affect the levels of p53 mRNA and protein in TMK‐1 and MKN‐28 cells, which contain mutated p53 genes. These mutated p53 complementary DNAs, when overexpressed, failed to activate transcription from the p21 promoter. Furthermore, TGFβ1 caused a reduction in the steady‐state level of cyclin A protein concomitantly with inhibition of cdk2 kinase activity in TMK‐1 cells. These results suggest that the growth inhibition of tumor cells by TGFβ1 is associated with p53‐independent induction of p21, subsequent suppression of cdk activity and a decrease in cyclin A protein in TMK‐1 cells.
Fibrotic Focus in Invasive Ductal Carcinoma: An Indicator of High Tumor Aggressiveness
Authors: Hasebe Takahiro , Tsuda Hitoshi , Hirohashi Setsuo , Shimosato Yukio , Iwai Minato , Imoto Shigeru , Mukai Kiyoshi
Published: August 2005

Histological examination of invasive ductal carcinoma of the breast often demonstrates the presence of an extensive central fibrotic focus (FF). The clinicopathological significance of the FF, or scar, in primary invasive ductal carcinoma is still ambiguous. One hundred and fifty‐three cases of invasive ductal carcinoma (IDC) were classified into two groups, those with and those without FF. The differences in frequency of immunohistochemically detected overexpression of c‐erb B‐2 protein and nuclear accumulation of p53 protein, and the labeling index of proliferating cell nuclear antigen (PCNA), as well as histopathological parameters were compared between these two groups. IDCs smaller than 50 mm with FF showed a higher frequency of high‐grade tumors, a higher frequency of lymph node metastasis, and a significantly higher frequency of c‐erb B‐2 protein overexpression than those without FF. In tumors of 20 mm or less, the incidence of nuclear accumulation of p53 protein was significantly higher in tumors with than those without FF. Tumors with FF showed a significantly higher PCNA labeling index than those without FF, regardless of tumor size. The present results indicate that the presence of FF is an important clinicopathological parameter associated with a higher degree of malignancy in IDCs, especially those smaller than 50 mm. Therefore, dividing IDCs into those with and those without FF appears to be meaningful clinicopathologically.
A Human/Mouse Chimeric Monoclonal Antibody against Intercellular Adhesion Molecule‐1 for Tumor Radioimmunoimaging
Authors: Yamamura Miyuki , Hinoda Yuji , Sasaki Shigeru , Tsujisaki Masayuki , Oriuchi Noboru , Endo Keigo , Imai Kohzoh
Published: August 2005

A mouse‐human chimeric antibody for intercellular adhesion molecule‐1 (ICAM‐1) was established by using heavy chain loss mouse mutant hybridoma and human immunoglobulin expression vector. The HA58 hybridoma secreted anti‐ICAM‐1 monoclonal antibody (MoAb) (IgG1, k). The gene of the mouse variable region of heavy chain was amplified and cloned by the polymerase chain reaction technique directly from the HA58 hybridoma RNA. The variable region of heavy chain was joined with an expression vector which contains human γ1 constant gene. The expression vector was transfected into heavy chain loss mutant cells HA58‐7, which produced only murine immunoglobulin light chains. The resultant chimeric MoAb HA58, chHA58, retained full‐binding reactivity to ICAM‐1 compared with murine HA58 parental antibody. The chimeric MoAb chHA58 showed little antibody dependent cell‐mediated cytotoxic activity against cultured tumor cells. Biodistribution studies with 99mTc‐labeled chHA58 in nude mice bearing human gastric carcinoma JRST cells demonstrated that the tumor‐blood ratio was 1.55 at 18 h after injection, when the tumors were clearly visible in gamma scintigraphy. These data suggest that chHA58 may be of practical use for radioimmunoimaging of a wide variety of tumors.
Murine Asialo GM1+CD8+ T Cells as Novel Interleukin‐12‐responsive Killer T Cell Precursors
Authors: Lee Ushaku , Santa Kazuki , Habu Sonoko , Nishimura Takashi
Published: August 2005

Freshly isolated CD8+ T cells, but not CD4+ T cells, contained 20–30% of asialo GM1+ (ASGM1+) T cells which were distinct from ASGM1+NK1.1+ natural killer cells. This novel ASGM1+CD8+ T cell subpopulation showed a strong proliferative response to interlenkin‐12 (IL‐12) in the presence of IL‐2. Culture of ASGM1+CD8+ T cells with IL‐12 plus IL‐2 allowed the generation of anomalous killer T cells concomitantly with the accumulation of cytolytic molecules. Moreover, ASGM1+CD8+ T cells produced high levels of interferon‐γ (IFN‐γ), but not IL‐4, upon stimulation with IL‐12 plus IL‐2. Such immune responses were not observed in ASGM1− CD8+ T cell snbpopulations constituting the majority of CD8+ T cells. These results demonstrated that ASGM1+CD8+ T cells are a novel subpopulation of IL‐12‐responsive and IFN‐γ‐producing killer T cell precursors.
Sinusoidal Capillarization and Arterial Blood Supply Continuously Proceed with the Advance of the Stages of Hepatocarcinogenesis in the Rat
Authors: Yamamoto Takatsugu , Kaneda Kenji , Hirohashi Kazuhiro , Kinoshita Hiroaki , Sakurai Masami
Published: August 2005

Hepatocellular carcinoma (HCC) is supplied only with arterial blood and has capillaries instead of sinusoids, unlike the normal hepatic tissue. In order to reveal the sequential changes of sinusoidal structure and blood supply during hepatocarcinogenesis, we examined hyperplastic nodules (HPN), atypical hyperplastic nodules (AHPN) and HCC in F344 rats fed with 2‐acetylaminofluorene for 25–35 weeks. The extent of arterial blood supply and the degree of sinusoidal capillarization were assessed by the staining of hepatic nodules with arterially infused ink and by immunohistochemical and ultrastructural analyses of the sinusoids, respectively. The proportion of arterialized nodules was 47% in HPN, 57% in AHPN and 85% in HCC, which indicates that arterialization begins at the stage of HPN and a few HCC still receive portal venous blood. The proportion of Factor‐VIII‐related antigen (F‐VIII)‐positive nodules was 25% in HPN, 40% in AHPN and 100% in HCC. Non‐arterialized nodules did not express F‐VIII, while some F‐VIII‐negative nodules were already arterialized. Electron microscopically, non‐arterialized HPN exhibited normal features of sinusoids, while arterialized HPN showed disappearance of Kupffer cells and partial defenestration of sinusoidal endothelial cells. In non‐arterialized and arterialized AHPN, basement membrane became continuous and lipid droplet‐containing stellate cells were no longer seen. In HCC, endothelial fenestrae were diminished and basement membrane became thick. The present study has demonstrated that sinusoids are altered in the following order as hepatocarcinogenesis advances from HPN to HCC; onset of arterialization and decrease of endothelial fenestrae, expression of F‐VIII by endothelial cells, disappearance of Kupffer cells, diminution of lipid droplets in stellate cells and thickening of basement membrane.
Frequent Microsatellite Instabilities and Analyses of the Related Genes in Familial Gastric Cancers
Authors: Akiyama Yoshimitsu , Nagasaki Hiromi , Nihei Zenro , Iwama Takeo , Nomizu Tadashi , Utsunomiya Joji , Yuasa Yasuhito
Published: August 2005

Microsatellite instability or replication error seems to be related to defective DNA mismatch repair genes, such as hMSH2, hMLH1, hPMS1 and hPMS2, which have been identified as causative genes of hereditary nonpolyposis colorectal cancers (HNPCC). Recently, it was reported that mutations at the simple repeated sequences in the transforming growth factor‐β type II receptor (TGF‐β RII) gene occurred in replication error‐positive colorectal cancers. To determine genetic alterations in familial gastric cancers (FGC), we examined replication error using eight microsatellite DNA markers, and screened mutations in the hMSH2, hMLH1 and TGF‐β RII genes in six cases from four FGC kindreds. Moreover, hMTH1, a human homolog of the bacterial mutT gene, was also screened. Four of six (67%) cancers showed the replication error‐positive phenotype, indicating that microsatellite instability is highly associated with not only HNPCC, but also FGC. No germline mutation was found in the whole coding sequences of hMSH2 and hMTH1, or in the conservative regions of hMLH1 in any patient, while one cancer DNA showed a somatic mutation at codon 682 (threonine to alanine) in hMSH2. No alteration was found at the small repeated sequences in TGF‐β RII in FGC tumor DNA. These results indicate that the carcinogenetic process of FGC may be different from that of HNPCC.
Genetic Controls of Susceptibility and Resistance to 4‐Nitroquinoline 1‐Oxide‐induced Tongue Carcinomas in Rats
Authors: Kitano Motoo , Hirayama Yoshikazu , Tanuma Jun‐ichi , Matsuuchi Hiroaki , Miura Yoshihiro , Li Tie‐Jun , Semba Ichiro , Ozaki Hiroki S. , Kokubu Teiji , Hatano Hiromichi , Tada Mariko , Kobayashi Yasuto , Shisa Hayase
Published: August 2005

We analyzed the incidence of infiltrative mass‐type tongue carcinomas (IMTC) induced in 550 rats by continuous oral administration of 0.001% 4‐nitroquinoline 1‐oxide solution for 180 days. The study included various crosses of susceptible Dark‐Agouti rats (DA) and resistant Wistar/Furth rats (WF). DA showed a 93.6% incidence of IMTC measuring more than 5 mm in their largest diameter, while WF showed only a 4% incidence. Reciprocal F1 and F2 hybrids mated by DA and WF showed 47.5% and 45.8% incidences, respectively. Meanwhile, reciprocal backcrossed hybrids to DA and WF showed 73.7%, and 24.6% incidences, respectively. Segregation of the incidences suggests that there are two autosomal dominant genes, one linked to the susceptibility of DA and the other to the resistance of WF.
The Multidrug Resistance‐associated Protein Gene Confers Drug Resistance in Human Gastric and Colon Cancers
Authors: Tomonaga Michio , Oka Mikio , Narasaki Fumihiko , Fukuda Minoru , Nakano Reiji , Takatani Hiroshi , Ikeda Koki , Terashi Kenji , Matsuo Isao , Soda Hiroshi , Cowan Kenneth H. , Kohno Shigeru
Published: August 2005

To determine the expression of multidrug resistance‐associated protein (MRP) gene and its role in gastric and colon cancers, we analyzed 10 gastric and 10 colon non‐drug‐selected cell lines and a similar number of tissue samples of these cancers. We compared the expression of MRP and mdr1 mRNA in cell lines and tissues using reverse‐transcriptase polymerase chain reaction. In mdr1‐negative cells, the relationship between the level of MRP gene expression and sensitivity to anticancer drugs was examined. The effect of verapamil, an MRP‐modulating agent, was also examined in these cells. The expression of MRP gene in gastric cancer cell lines varied from a low to a high level, but mdr1 was not detected in any of these cell lines. Colon cancer cell lines expressed low to intermediate levels of MRP gene, and half of the cells co‐expressed low to high levels of mdr1. In tissue samples, the expression pattern of the two multidrug resistance (MDR) genes was broadly similar to that described for the cell lines, except that most of the gastric cancer tissue samples did express low levels of mdr1. No significant correlation was observed between the level of MRP gene expression and sensitivity to anticancer drugs in gastric and colon cell lines. However, verapamil significantly increased the sensitivity to etoposide, doxorubicin and vincristine in cells highly expressing MRP gene. Our results indicate that MRP gene may be important in conferring MDR in gastric and colon cancer cells.
Increased Radiosensitivity of p16 Gene‐deleted Human Glioma Cells after Transfection with Wild‐type p16 Gene
Authors: Miyakoshi Junji , Kitagawa Kaori , Yamagishi Nobuyuki , Ohtsu Shuji , Day Rufus S. , Takebe Hiraku
Published: August 2005

The A1235 and T98 cell lines derived from human gliomas have homozygous deletions in their p16 genes and are radiosensitive and radioresistant, respectively, with respect to other established glioma cell lines. These differences in radiosensitivity may be due to variations to some extent among cell lines, rather than genetically defined resistance or sensitivity. We examined the effect on radiation sensitivity of introducing a wild‐type p16 gene into both p16‐deficient glioma cell lines. The plasmid pOPMTS containing human wild‐type plfi cDNA and a neomycin resistance gene, or the control plasmid pOPRSV1, were transfected into these cells. Clones from both cell lines, which expressed wild‐type p16 mRNA constitutively after transfection with pOPMTS, were more radiosensitive than the parental cells and clones obtained after transfection with the negative control plasmid.
Germline Mutations of the MEN1 Gene in Japanese Kindred with Multiple Endocrine Neoplasia Type 1
Authors: Shimizu Satoko , Tsukada Toshihiko , Futami Hitoyasu , Ui Kotaro , Kameya Toru , Kawanaka Masahiro , Uchiyama Shozo , Aoki Akira , Yasuda Hiroko , Kawano Shin‐ichiro , Ito Yukio , Kanbe Masako , Obara Takao , Yamaguchi Ken
Published: August 2005

Multiple endocrine neoplasia type 1 (MEN1) is an autosotnal dominant familial cancer syndrome. The responsible gene MEN1 has recently been isolated, and its germline mutations have been identified in affected individuals in the United States, Canada and Europe. We screened for MEN1 mutations by direct nucleotide sequencing of all protein‐coding regions, and identified five distinct germline mutations in five among six Japanese kindreds with familial MEN1 or familial hyperparathyroidism. The mutations were dispersed across the gene. These findings suggest that, because of the absence of an obvious founder effect, the entire MEN1 gene region should be examined for germline mutations in the probands of MEN1 and related syndromes in Japanese families.
Transforming Growth Factor‐β and Hepatocyte Growth Factor Produced by Gastric Fibroblasts Stimulate the Invasiveness of Scirrhous Gastric Cancer Cells
Authors: Inoue Tohru , Chung Yong‐Suk , Yashiro Masakazu , Nishimura Shigehiko , Hasuma Tadayoshi , Otani Shuzo , Sowa Michio
Published: August 2005

Scirrhous gastric carcinoma is characterized by cancer cells that infiltrate rapidly in the stroma with extensive growth of fibroblasts. In the present study, we examined the effect of gastric fibroblasts on the invasiveness of a Scirrhous gastric cancer cell line, OCUM‐2D, using an invasion assay. Gastric fibroblast‐derived conditioned medium (CM) significantly stimulated the invasiveness of OCUM‐2D cells, as did transforming growth factor‐β (TGF‐β) and hepatocyte growth factor (HGF). The stimulating activity of gastric fibroblast‐derived CM was inhibited significantly by anti‐TGF‐β neutralizing antibody or anti‐HGF neutralizing antibody. TGF‐β and HGF were detected in the gastric fibroblast‐derived CM, and TGF‐β receptor and C‐met (HGF receptor) were expressed on OCUM‐2D cells. Thus, TGF‐β and HGF produced by gastric fibroblasts appear to affect the invasiveness of scirrhous gastric cancer cells. TGF‐β was also detected in the conditioned medium derived from OCUM‐2D cells, though HGF was not. TGF‐β appears to affect the invasiveness of OCUM‐2D cells in both paracrine and autocrine fashions.
Mutational Analysis of BRCA1 Gene in Ovarian and Breast‐ovarian Cancer Families in Japan
Authors: Takano Masashi , Aida Hiroshi , Tsuneki Ikunosuke , Takakuwa Koichi , Hasegawa Isao , Tanaka Hajime , Saito Masaaki , Tsuji Shoji , Sonoda Takahiko , Hatae Masayuki , Chen Jui‐Tung , Takahashi Katsuyuki , Hasegawa Kazuo , Toyoda Nagayasu , Saito Noriyasu , Yakushiji Michiaki , Araki Tsutomu , Tanaka Kenichi
Published: August 2005

We analyzed the alteration of BRCAI in DNA obtained from 83 individuals of 13 Japanese site‐specific ovarian cancer families and 6 breast‐ovarian cancer families. Six germline mutations were detected in 7 families, which consisted of 4 breast‐ovarian cancer and 3 site‐specific ovarian cancer families, by single‐strand conformation polymorphism analysis, followed by direct sequence determination. The mutations included three framcshifts, two nonsense mutations, and one missense mutation causing loss of a zinc‐binding motif. The frequency of loss of heterozygosity at the microsatellite markers on the BRCAI gene was 57% (8 of 14 cases) in site‐specific ovarian cancer families, and 100% (6 of 6 cases) in breast‐ovarian cancer families. All tumors of the patients carrying a mutation of BRCAI showed deletion of wild‐type alleles, implicating BRCAI as a tumor suppressor gene. Tbese results suggest tbat germline mutations of the BRCAI gene play an important role in the carcinogen‐esis of breast and/or ovarian cancer in a majority of breast‐ovarian cancer families and in some site‐specific ovarian cancer families.
Modulation of Motility and Proliferation of Glioma Cells by Hepatocyte Growth Factor
Authors: Yamamoto Shinji , Wakimoto Hiroaki , Aoyagi Masaru , Hirakawa Kimiyoshi , Hamada Hirofumi
Published: August 2005

Invasive proliferation is a critical biological characteristic of gliomas. We evaluated the activities of hepatocyte growth factor (HGF) on proliferation and motility of glioma cells, comparing them with the effects of other growth factors (EGF, bFGF, PDGF‐BB, TGF‐βl). Seven primary culture lines all expressed c‐met and HGF mRNA, and secreted HGF. HGF stimulated 3H‐thymidine uptake of every glioma cell line (30 to 70% upregulation). Boyden chamber assay and scattering assay revealed that HGF promoted cell motility with chemokinetic and strong chemotactic activities. Concentric circle assay showed that HGF promoted two‐dimensional expansion (proliferation and motility) most strongly among the growth factors studied. Further, we analyzed 23 paraffin‐embedded sections of surgically resected gliomas (7 grade II, 8 grade III, and 8 grade IV) by immunohistochemistry. Expression of HGF and Met increased with malignant progression of gliomas, suggesting that gliomas stimulated their invasive proliferation by autocrine HGF production. Neurons and vasculature were HGF‐positive, and Met‐positive glioma cells gathered around them. The data indicate that neurons and vasculature, which are the main tracks of glioma invasion, augment chemotactic invasion and proliferation of gliomas by paracrine HGF secretion. Clearly HGF plays a critical role in invasive proliferation of glioma cells and it is therefore a candidate target of therapeutic intervention.
Two Distinct Human Uterine Cervical Epithelial Cell Lines Established after Transfection with Human Papillomavirus 16 DNA
Authors: Ohta Yujiro , Tsutsumi Kouichiro , Kikuchi Keiji , Yasumoto Shigeru
Published: August 2005

We have established two distinct human cervical cell lines, NCC16 and NCE16, after traiisfecting human papillomavirus type 16 (HPV16) DNA into normal human ecto‐cervical and endo‐cervical epithelial cells, respectively. Both lines expressed HPV16 E6 and E7 as detected hy reverse transcriptase‐polymerase chain reaction and northern Wot hybridization. These cells have been passaged for over 100 population doublings and express strong telomerase activity. Neither cell line was tumorigenic in athymic nu/nu mice. However, both NCC16 and NCE16 developed abnormally stratified architectures following implantation with a silicon membrane sheet in the back of athymic nude mice. The former cells were pathohistologically similar to carcinoma, while the latter produced Alcian‐blue positive cells, suggesting the occurrence of metaplastic changes. These distinct cell lines offer a useful model system for the study of cervical carcinogenesis and of its regulatory mechanism after HPV infection in different regions of the uterine cervix.
Quantitative Analysis of Cyclin Dl Messenger RNA Expression in Head and Neck Squamous Cell Carcinomas
Authors: Monden Nobuya , Nishizaki Kazunori , Fukushima Kunihiro , Masuda Yu , Tsukuda Kazunori , Shimizu Kenji
Published: August 2005

Cyclin Dl is thought to play a critical role in the G1/S phase transition of the cell cycle. Amplification of this gene has been reported in several types of human neoplasms including breast, lung, esophageal, and head and neck tumors. In this study, we have analyzed the relative level of expression of cyclin D1 messenger RNA (mRNA) in fresh specimens of head and neck squamous cell carcinoma (HNSCC), and investigated the concordance of the overexpression of cyclin D1 mRNA with gene amplification. Levels of cyclin D1 mRNA were analyzed hy a modified method of competitive reverse transcription‐polymerase chain reaction and levels of cyclin D1 gene amplification were evaluated by Southern blot hybridization in a series of 23 matched normal mucosas and HNSCC. Overexpression of cyclin D1 mRNA was observed in 10 of 23 cases (43.5%) of HNSCC, ranging from 2 to 50‐fold higher than the normal control. Twelve of 23 cases could be evaluated by Southern blot hybridization, and gene amplification was found in only 2 of 12 cases (16.7%), These findings suggest that cyclin D1 plays an important role in tumorigenesis of HNSCC, and gene amplification is not one of the major mechanisms for overexpression of cyclin D1.
Inhibitory Effects of Diallyl Bisulfide or Aspirin on 2‐Amino‐l‐methyl‐6‐phenylimidazo[4,5‐b]pyridine‐induced Mammary Carcinogenesis in Rats
Authors: Suzui Natsuko , Sugie Shigeyuki , Rahman K. M. Wahidur , Ohnishi Masami , Yoshimi Naoki , Wakabayashi Keiji , Mori Hideki
Published: August 2005

Modifying effects of diallyl disulfide (DAD), aspirin or DL‐α‐difluoromethylornithine (DFMO) on 2‐amino‐l‐methyl‐6‐phenyIimidazo[4,5‐b]pyridine (PhlP)‐induced mammary carcinogenesis in SD rats were investigated. A total of 166 female rats, 6 weeks old, were divided into 8 groups. They were fed a high fat diet throughout the experiment. Starting at 7 weeks of age, groups 1–4 were given PhIP (85 rag/kg body weight in corn oil) by gavage 8 times in 10 days, and groups 58 were given corn oil alone. For the beginning 4 weeks, groups 2 and 5 were given DAD at 200 ppm in diet. Similarly groups 3 and 6, and groups 4 and 7 were given aspirin (400 ppm) and DFMO (400 ppm), respectively. Mammary carcinomas were only recognized in groups 1–4 at the termination (25 weeks after the start of experiment). Multiplicity (mean number/rat) of neoplasms in group 2 (PhIP+DAD, 0.90/rat) and group 3 (PhIP+aspirin, 1.37/rat) was significantly smaller than that in group 1 (PhIP alone, 2.457 rat) (P < 0.005 and P < 0.05, respectively). These results indicate that dietary intake of DAD or aspirin during the time corresponding to initiation phase has chemopreventive potential on PhlP‐induced mammary carcinogenesis in rats.
Telomerase Activity and Metastasis: Expansion of Cells Having Higher Telomerase Activity within Culture Lines and Tumor Tissues
Authors: Saito Yoshiyuki , Kosugi Shin‐ichi , Suda Takeshi , Wakabayashi Yuichi , Mishima Yukio , Hatakeyama Katsuyoshi , Kominami Ryo
Published: August 2005

Tumor cells with metastatie potential may have a high telomerase activity that augments telomeric DNA repeats, allowing the cells to escape from the inhibition of cell proliferation due to shortened telomeres. We examined the expression level of telomerase activity using the telomeric repeat amplification protocol among a series of cell lines obtained by repeated transplantation of a mouse fibrosarcoma. The lines could be grouped into three; one has no metastatic potential, and the other two show metastatic abilities after intravenous or subcutaneous injection. Comparison of their telomerase activity indicated that more malignant lines had higher activity. A similar relation was seen in metastatic nodules formed through clonal expansion from the heterogeneous population of inoculated cells; clonality was monitored in terms of variable patterns of subtelomeric repeats. The results suggest that a high level of telomerase activity may not be requisite for metastasis, but may confer a propensity to dominate in a tumor tissue.
Association for Medical Education and Research in Substance Abuse
Authors: Samet Jeffrey H. , Galanter Marc , Bridden Carly , Lewis David C.
Published: August 2005

The Association for Medical Education and Research in Substance Abuse (AMERSA) is a multi‐disciplinary organization committed to health professional faculty development in substance abuse. In 1976, members of the Career Teachers Training Program in Alcohol and Drug Abuse, a US federally funded multi‐disciplinary faculty development program, formed AMERSA. The organization grew from 59 founding members, who were primarily medical school faculty, to over 300 health professionals from a spectrum of disciplines including physicians, nurses, social workers, dentists, allied health professionals, psychologists and other clinical educators who are responsible for advancing substance abuse education. AMERSA members promote substance abuse education among health professionals by developing curricula, promulgating relevant policy and training health professional faculty to become excellent teachers in this field. AMERSA influences public policy by offering standards for improving substance abuse education. The organization publishes a peer‐reviewed, quarterly journal, Substance Abuse, which emphasizes research on the education and training of health professions and also includes original clinical and prevention research. Each year, the AMERSA National Conference brings together researchers and health professional educators to learn about scientific advances and exemplary teaching approaches. In the future, AMERSA will continue to pursue this mission of advancing and supporting health professional faculty who educate students and trainees to address substance abuse in patients and clients.
Comparing the effectiveness of two types of cooling blankets for febrile patients
Authors: Loke Alice Y , Chan Helen CL , Chan Tony MF
Published: August 2005

A randomized controlled trial was conducted to compare the effectiveness of airflow‐ and waterflow‐cooling blankets for febrile patients in intensive care unit. More patients in the airflow than in the waterflow‐cooling blanket group reached the desired temperature of <38°C at the end of 8 h (94·1 versus 60·0%, χ2 = 2·86, P = 0·047). Patients in the airflow blanket group also took less time than those in the waterflow blanket group to achieve the desired temperature (3·1 and 5·7 h, respectively, U = 2·00, P < 0·001). These findings suggest that the airflow blanket is the cooling blanket of choice for febrile patients when external cooling is needed.
The measurement and application of thrombin generation
Authors: Baglin Trevor
Published: August 2005

It is the capacity to generate thrombin, and the enzymatic work that thrombin does, that determines blood coagulability. Therefore, measurement of the enzymatic work potential of thrombin provides a method for quantifying the composite effect of the multiple factors that determine coagulation capacity. The application of measurement of thrombin generation to clinical decision making has been hampered by numerous technical difficulties and pitfalls, many of which have now been overcome. Technical advances now permit rapid, reproducible measurement. A review of clinical studies performed to date indicates the need to appreciate the precise methodology used in each case and the need to consider standardisation in future studies. Applying thrombin generation measurement to clinical decision making will require up‐to‐date estimates of risks relating to the disorder and the intended therapeutic intervention. Ultimately management studies will be required if the clinical utility of measurement of thrombin generation is to be proven.
Indirubin, a Chinese anti‐leukaemia drug, promotes neutrophilic differentiation of human myelocytic leukaemia HL‐60 cells
Authors: Suzuki Kazuhiro , Adachi Reiko , Hirayama Akiko , Watanabe Hidemi , Otani Saki , Watanabe Yuka , Kasahara Tadashi
Published: August 2005

Indirubin, a purple vegetable dye, is a traditional Chinese medicine for myelocytic leukaemia. Indirubin inhibits cyclin‐dependent protein kinases (CDKs) and is present in human urine and serum. When indirubin was present during the neutrophilic differentiation of human myelocytic leukaemia HL‐60 cells, it augmented superoxide production triggered by opsonized zymosan (OZ) by the terminally differentiated HL‐60 cells. It also augmented the calcium response to OZ stimulation, and HL‐60 cell chemotaxis evoked by interleukin‐8 (IL‐8, CXCL8) and formylpeptide. In addition, indirubin induced marked IL‐8 release by the cells during differentiation and the cells differentiated with indirubin had typical neutrophilic properties, deformed nuclei and granules. Use of stable cloned HL‐60 cells that contained a reporter vector for monitoring the activity of the transcription factor PU.1, which acts specifically at the stage of promyelocyte differentiation into neutrophils and monocytes, revealed that indirubin has a potent promoting activity on intracellular PU.1. Indirubin enhanced the expression of typical neutrophil proteins, including granulocyte‐colony stimulating factor receptor, the β2‐integrin subunit CD18, the NADPH‐oxidase subunit p47phox, and the IL‐8 receptor CXCR1, all are controlled by PU.1. Indirubin also inhibited CDK2‐dependent phosphorylation of retinoblastoma protein during neutrophilic differentiation. These results suggest that indirubin augments the neutrophilic differentiation of human myelocytic leukaemia HL‐60 cells through inhibition of CDK2 and activation of PU.1.