bluuebunny/crossref_metadata_2025 huggingface dataset:
ISO 14001 and its role in environmental preservation
Authors: Beal Susan
Published: October 2002

The ISO 14001 Environmental Management System (EMS) standard, which provides the course of action
for an organization to follow in its development of an EMS, is based on a Plan, Do, Check, Act cycle that fosters
the continual improvement of environmental preservation goals. This incremental process begins when an
organization first recognizes how its practices affect the environment, identifies how it can develop the
programs that will allow management of its own unique “significant environmental aspects,” and
determines how to best adhere to applicable environmental rules and regulations. Each organization will find that
the implementation of ISO 14001 provides better control over its processes, resulting in the minimization of
out‐of‐compliance issues. In particular, integrating environmental management with business
operations allows an organization to better manage the environmental impacts of those operations. Moreover, by
identifying and documenting operational controls as part of the EMS process, an organization cannot help but
achieve continual environmental improvement. A documented EMS is especially important because it functions to
increase employee awareness of what is required to achieve environmental goals, as well as enables the evaluation
of the system through internal assessments. Accomplishing the above tasks leads to environmentally sound
performance and, in turn, to the readiness that ensures achievement of an organization's primary
mission—to provide efficient and affordable services to its customers. Although the outcome of implementing
an EMS in accordance with the ISO 14001 standard considerably improves the effectiveness and efficiency of an
organization's environmental program, only those organizations that have fully implemented the requirements
outlined in the standard are officially registered to ISO 14001. This paper is presented to describe the
implementation of the ISO 14001 standard at the Naval Undersea Warfare Center Division, Newport, with emphasis on
the benefits that were gained and the lessons that were learned. Division Newport was the first Department of
Defense activity to register an entire federal facility to the ISO 14001 standard, and received the 2001 White
House Closing the Circle Award in the EMS category. © 2002 Wiley Periodicals, Inc.
Laminar diffusion of suspended particulate matter using a two phase flow model
Authors: Panda T. C. , Mishra S. K. , Panda K. Ch.
Published: October 2002

The present paper envisages laminar mixing of a two‐dimensional jet of particulate suspension in an incompressible carrier fluid with a free stream in direction of the jet axis. Finite difference technique has been employed for finding out solution of governing equations. It is found that the diffusion parameter ε, the ratio of particle diffusion coefficient and kinematic viscosity of the carrier fluid, have significant influence on the concentration of particles. A large value of ε has the effect in increasing the perturbation velocity up and perturbation density ρp. It is observed that the volume fraction φ, has no significant effect on perturbation velocity u and up but has profound effect on perturbation velocity v and vp. It is also found that the particle phase as well as the carrier fluid velocity attain free stream value for the large ξ, the modified x‐co‐ordinate. Further the magnitude of the perturbation quantities u, up, v, vp decreases as ξ increases i.e. at far away from the nozzle exit. Copyright © 2002 John Wiley & Sons, Ltd.
Zonal multi‐domain RANS/LES simulations of turbulent flows
Authors: Quéméré P. , Sagaut P.
Published: October 2002

A new zonal multi‐domain Reynolds averaged numerical simulation/large‐Eddy simulation (RANS/LES) method is presented and assessed. The main occurring problem is the coupling at the domain interfaces between a 1D or 2D, ensemble averaged solution computed with the RANS approach with the 3D, filtered, unsteady solution obtained using the LES approach. Both low‐ and high‐normal velocity interface cases are considered, resulting in a fully general approach. A coupling procedure well suited for cell‐centred finite volume numerical method is proposed. Numerical tests are carried out for the plane channel and the plane plate with a blunt trailing‐edge configurations. Copyright © 2002 John Wiley & Sons, Ltd.
Gemcitabine following radiotherapy with concurrent 5‐fluorouracil for nonmetastatic adenocarcinoma of the pancreas
Authors: Kachnic Lisa A. , Shaw James E. , Manning Matthew A. , Lauve Andrew D. , Neifeld James P.
Published: October 2002

Gemcitabine has been shown to be an active agent in the treatment of pancreatic cancer. This study was conducted to prospectively examine the tolerance and early efficacy of adjuvant gemcitabine following radiotherapy with concurrent 5‐fluorouracil (5‐FU) for nonmetastatic pancreatic adenocarcinoma. Twenty‐three patients, median age 64 years, were treated with combined modality therapy. Nine patients underwent tumor resection before chemoradiation; 14 patients with locally unresectable tumors received definitive chemoradiation. Radiotherapy utilized four fields to the tumor and lymphatics to 45 Gy, plus a lateral boost to 50.4 Gy. Concurrent 5‐FU 500 mg/m2/day was administered on days 1–3 and 29–31, followed by 4 months of gemcitabine 1,000 mg/m2/week for 3 weeks (fourth week break). Adjuvant gemcitabine was well tolerated. Eighty‐three percent of the patients completed three to four cycles. The primary dose‐limiting toxicity was leukopenia, which was observed in 10 patients (43%). Nonhematologic toxicities were reported in five patients (22%). There were no cases of gemcitabine‐induced radiation recall and there have been no deaths attributed to treatment toxicity. Median follow‐up for the 23 patients was 12 months (range, 5–50); the actuarial median survival was 13 months. This report confirms that adjuvant gemcitabine following radiotherapy with concurrent 5‐FU for nonmetastatic pancreatic adenocarcinoma can be safely administered. © 2001 Wiley‐Liss, Inc.
Non‐human to human organ transplantation:Its biologic basis and a potential role for radiation therapy
Authors: Halperin Edward C.
Published: October 2002

There is an inadequate supply of human donor organs for transplantation. Xenotransplantation, the transplantation of organs from non‐human animals to humans, is one of the potential solutions to this problem. The pig appears to be the preferred donor. For xenotransplantation to be successful, researchers must deal with three fundamental problems: (1) Hyperacute rejection of porcine organs, related to binding of xenoreactive natural antibodies of the recipient to antigens on the graft's endothelial cells, must be overcome. (2) Transmission of animal pathogens to humans must be prevented. Concern about zoonosis is not only directed to the transplant recipient but also concerns the risk that an infectious agent will be transferred from the recipient to the general population. (3) The xenografted organ must be physiologically compatible with the recipient. The physiological function of a pig organ in a human and its ability to sustain a human are problematic. Total lymphoid irradiation (TLI) and thoraco‐abdominal irradiation (TAI) as immunosuppressive modalities have been investigated in rodent‐to‐rodent, large mammals and non‐human primates‐to‐primates, and pig‐to‐primate models. In certain clinical situations, TLI and TAI may prove to be important components for the preparation of the xenotransplant recipient. Progress in genetic engineering and cloning may soon lead to clinical trials in xenotransplantation. © 2001 Wiley‐Liss, Inc.
Expression of hyaluronan synthase 2 or hyaluronidase 1 differentially affect the growth rate of transplantable colon carcinoma cell tumors
Authors: Jacobson Annica , Rahmanian Mehdi , Rubin Kristofer , Heldin Paraskevi
Published: October 2002

Advanced colorectal cancers are often associated with elevated amounts of hyaluronan. To investigate the importance of hyaluronan in colon carcinoma tumor progression, we have expressed by stable transfection hyaluronan synthase 2 (Has2) and hyaluronidase 1 (Hyal1) in the rat colon carcinoma cell line, PROb. We found that hyaluronan overproduction led to a higher growth rate of tumor cells in vitro, and to a faster development of transplantable tumors in syngeneic rats, compared to the mock‐transfectants. Has2 transfected PROb cells gave rise to tumors that were significantly less vascularized, but had a significantly larger viable tumor fraction compared to tumors generated from mock‐transfectants. In contrast, Hyal1 overexpression suppressed the growth rate of tumor cells both in vitro and in vivo. Moreover, tumors derived from Hyal1‐transfected cells had a significantly larger necrotic area than tumors derived from mock‐ and Has2‐transfectants. Our study demonstrates that Has2 overproduction promotes tumorigenicity, whereas Hyal1 overexpression suppresses tumorigenicity in an experimental model for colon carcinoma. © 2002 Wiley‐Liss, Inc.
Type III intestinal metaplasia and carcinoma express an identical carbohydrate‐epitope revealed by a novel monoclonal antibody (2D11)
Authors: Kimura Akihiko , Ohta Masataka , Ono Kazuo , Yamaguchi Kazuya , Inoue Hiroshi , Yasuda Seiji , Nosaka Mizuho , Tsuji Tsutomu
Published: October 2002

A monoclonal antibody (2D11, IgG2b) obtained by immunizing mice with a mucin fraction of the human gastric mucosa reacted specifically to intestinal metaplasia of human gastric mucosa and fetal intestinal mucosa but not to normal adult gastric, small intestinal or colonic mucosa in immunohistochemical staining. The results of Western blotting indicated that 2D11 recognized the high molecular weight glycoprotein(s) (mucin) of the stomach. Treatment of the antigens with sodium periodate abolished their reactivity to 2D11, and digestion of the antigens with β‐galactosidase reduced their reactivity to 2D11. Digestion of the antigens with pronase had no effect, however, suggesting that 2D11 recognizes the oligosugar moiety but not the peptide moiety of the antigens. Further immunohistochemical investigation showed that the reactivity of 2D11 was restricted to the Type III intestinal metaplasia that is identified by a characteristic staining pattern with the high iron diamine‐Alcian blue stain. 2D11 also reacted in high frequency to adenocarcinomas of the stomach (66.7%), pancreas (66.7%) and gallbladder (50.0%), but in low frequency to those in lung (8.3%) and colon (11.1%). It is of interest that 2D11 reacted to very restricted regions of the gastric adenocarcinomas. All monoclonal antibodies to mucin polypeptides (MUC1, 2, 3, 5AC and 6) examined stained intestinal metaplasia and carcinomas in a different pattern from 2D11 in immunohistochemistry. These facts indicate that Type III intestinal metaplasia and carcinomas express carbohydrate chains identical to those expressed in the fetal intestinal mucosa, suggesting that both of them are closely related to fetal intestinal mucosa. © 2002 Wiley‐Liss, Inc.
Reduced risk of prostate cancer among patients with diabetes mellitus
Authors: Weiderpass Elisabete , Ye Weimin , Vainio Harri , Kaaks Rudolf , Adami Hans‐Olov
Published: October 2002

Although diabetes mellitus is associated with an increased risk of several malignancies, a negative association with prostate cancer is biologically most plausible. The epidemiologic evidence is, however, inconsistent, limited and based mostly on small studies. We present results from a large, population‐based cohort study in Sweden, where we assessed prostate cancer risk among patients hospitalized for diabetes mellitus. The cohort was composed of patients identified in the Swedish In‐Patient Register as having a hospital discharge diagnosis of diabetes mellitus in 1965–1994. The follow‐up was done by linkages with the national cancer register and other population‐based registers. Standardized incidence ratios (SIRs), with 95% confidence interval (CI), were used as a measure of relative risk. After complete exclusion of the first year of follow‐up (to avoid selection bias), 135,950 men remained in the cohort, contributing 827,099 years of follow‐up to the study. A total of 2,455 incident cases of primary prostate cancer were identified during 1–31 years of follow‐up, yielding an overall SIR of 0.91 (95% CI 0.87–0.94); this risk reduction was more pronounced among patients who have been hospitalized for diabetic complications (SIR = 0.82; 95% CI 0.74–0.91). We found no consistent trends in risk related to age at first hospitalization or to duration of follow‐up. We did find a small, but significantly decreased risk of prostate cancer among men who had been hospitalized for diabetes mellitus. © 2002 Wiley‐Liss, Inc.
Poorly differentiated adenocarcinoma and mucinous carcinoma of the colon and rectum show higher rates of loss of heterozygosity and loss of E‐cadherin expression due to methylation of promoter region
Authors: Kanazawa Takamitsu , Watanabe Toshiaki , Kazama Shinsuke , Tada Toshihiro , Koketsu Shinichiro , Nagawa Hirokazu
Published: October 2002

Previous studies have reported that poorly differentiated adenocarcinoma and mucinous carcinoma of the colon and rectum (Por & Muc) show a low incidence and poor prognosis. However, genetic alterations and tumourigenesis of Por & Muc remain unclear. In our study, we analyzed the genetic and epigenetic alterations of Por & Muc to clarify the difference from those of well‐differentiated adenocarcinoma (WD). First, we evaluated the loss of heterozygosity (LOH) on 4 chromosomes (2p, 5q, 17p, 18q) frequently observed in colorectal cancer. Second, to determine large‐scale allelic losses, we performed wide‐ranging allelotyping study. Using 27 microsatellite markers spanning every 10 cM on chromosome 17 and 18, we defined the LOH ratio as the proportion of markers that exhibit LOH out of 27 markers. Third, we evaluated the methylation of E‐cadherin. With respect to LOH of 4 loci, higher rates of LOH were observed in Por & Muc and a statistical significance was found on the markers adjacent to MSH2 and SMAD4/DCC. Moreover, as the evidence of large‐scale allelic imbalance, the average LOH ratio was higher in Por & Muc (0.70) than in WD (0.24) (p = 0.01). Methylation analysis showed that 54.5% of Por & Muc demonstrated hypermethylation of E‐cadherin. In immunohistochemistry, 77.8% of Por & Muc exhibited abnormal expression of E‐cadherin. Hypermethylation of E‐cadherin correlated significantly with abnormal expression (p = 0.047). These results suggest that the higher rates of LOH contribute to Por & Muc tumourigenesis and most of Por & Muc lose normal E‐cadherin expression, partly because of the methylation of promoter region. © 2002 Wiley‐Liss, Inc.
Dietary exposure and urinary excretion of total N‐nitroso compounds, nitrosamino acids and volatile nitrosamine in inhabitants of high‐ and low‐risk areas for esophageal cancer in southern China
Authors: Lin Kun , Shen Wenying , Shen Zhongying , Wu Yongning , Lu Shihhsin
Published: October 2002

We assessed the exposure of total N‐nitroso compounds (TNOCs) in the inhabitants of high‐ and low‐risk areas for esophageal cancer in southern China. Samples of 24 hr diet and 12 hr overnight urine were collected from 120 male adults in each of the 2 areas, a high‐risk area (Nan'ao County) and a low‐risk area (Lufeng County) for esophageal cancer. Annual standardized mortality rates of esophageal cancer in Nan'ao and Lufeng are 110/106 and 10/106 respectively. The 240 healthy male subjects (35–64 years old) were selected by a 3‐stage random cluster sample procedure. Levels of TNOCs, NAAs and volatile nitrosamines in the samples were measured. The TNOC detection rate (95%) in the diet, the TNOC daily intake (4.25 ± 0.84 μmol), TNOC excretion levels (0.04 ± 0.01 nmol/12 hr) and daily intake of volatile nitrosamines (5.84 ± 0.71 μmol) in the high‐risk area were significantly greater than values in the low‐risk area (A ± B = mean ± SE). The TNOC detection rate in the diet, the TNOC daily intake, TNOC excretion levels and daily intake of volatile nitrosamines in the low‐risk area were 70%, 0.25 ± 0.06 μmol, 0.02 ± 0.01 nmol/12 hr and 3.18 ± 0.31 μmol, respectively. NAA excretion levels showed no difference between the 2 areas (16.3 ± 7.18 μmol/12 hr for Nan'ao and 31.2 ± 26.4 μmol/12 hr for Lufeng). Thus, TNOCs are implicated in the etiology of esophageal cancer in southern China. © 2002 Wiley‐Liss, Inc.
Inhibition of prostate‐specific membrane antigen (PSMA)‐positive tumor growth by vaccination with either full‐length or the C‐terminal end of PSMA
Authors: Kuratsukuri Katsuyuki , Sone Tomomichi , Wang Ching Y. , Nishisaka Nobuyasu , Jones Richard F. , Haas Gabriel P.
Published: October 2002

For experimental immunotherapy of prostate cancer, we used a model system to target a defined region of the extracellular domain of prostate‐specific membrane antigen (PSMA). PSMA is a surface antigen expressed by prostate epithelium that is upregulated approximately 10‐fold in most prostate tumors. We vaccinated BALB/c mice with NIH3T3 cells cotransfected with pST/neo plus pEF‐BOS‐based vectors expressing either the full‐length 750–amino acid human PSMA or only the C‐terminal 180–amino acid region (PSMc). PSMc lies C‐terminal to the transferrin receptor‐like sequence in the extracellular domain of PSMA. BALB/c mice were injected i.p. 4 times at weekly intervals with vaccine cells. Vaccinated mice were then challenged s.c. with Renca/PSMA, a BALB/c renal cell carcinoma line transfected to express human PSMA. Growth of Renca/PSMA tumors was substantially retarded and host survival significantly prolonged in mice prevaccinated with either 3T3/PSMA or 3T3/PSMc. Furthermore, antiserum from vaccinated mice intensely immunocytochemically stained LNCaP, a PSMA‐positive human prostate cancer cell line. In contrast, control mice similarly prevaccinated i.p. with 3T3/neo (NIH3T3 cells transfected with pST/neo alone) developed Renca/PSMA tumors, which were palpable within 2 weeks and lethal by 5 weeks. Serum from 3T3/neo‐vaccinated mice did not immunocytochemically stain LNCaP cells. The antitumor activity induced by vaccination with 3T3/PSMc was also demonstrated via growth inhibition of established LNCaP tumors xenografted in athymic mice following passive transfer of immune serum from vaccinated mice. Our results suggest that vaccination with PSMc induces adaptive humoral activity, which is directed against the extracellular region of human PSMA and can significantly inhibit human prostate cancer growth in athymic mice, and that administration of antibodies to PSMA may provide a passive treatment modality for immunocompromised patients. © 2002 Wiley‐Liss, Inc.
Ectopic expression of herpes simplex virus‐thymidine kinase gene in human non‐small cell lung cancer cells conferred caspase‐activated apoptosis sensitized by ganciclovir
Authors: Chiu Chien‐Chih , Kang Yuan‐Lin , Yang Tsung‐Hsiang , Huang Chih‐Hsin , Fang Kang
Published: October 2002

Human non‐small cell lung cancer (NSCLC) cells were transfected with recombinant prodrug herpes simplex virus type I thymidine kinase (HSV‐tk) cDNA, and the selected clones underwent apoptosis in response to induction by antiviral ganciclovir (GCV). The efficiency of GCV‐induced growth inhibition and the extent of the bystander effect were associated with the expression level of HSV‐TK in stable transfectants. Development in the HSV‐tk/GCV system toward cell death was initiated with cell‐cycle accumulation at S and G2/M phases, immediately followed by the appearance of sub‐G0/G1 cells after drug exposure. To investigate the regulation of cell‐cycle modulators during drug treatment, we analyzed release of the apoptosis initiator cytochrome c and activation of the downstream effectors caspase‐9, caspase‐3 and poly(ADP‐ribose)polymerase 16 hr after GCV sensitization, followed by transient escalation of tumor‐suppressor p53 and cell‐cycle modulators cyclin A and B1 before committing to programmed cell death. Furthermore, tumor regression was proportional to the degree of ectopic expression of the transferred HSV‐tk gene. Our results demonstrate that the HSV‐tk/GCV system effectively inhibits the proliferation of NSCLC cells in vitro and in vivo through potent induction of apoptosis, thus providing a rationale for further development. © 2002 Wiley‐Liss, Inc.
Monocyte chemoattractant protein‐1 expression correlates with macrophage infiltration and tumor vascularity in human esophageal squamous cell carcinomas
Authors: Ohta Masahiro , Kitadai Yasuhiko , Tanaka Shinji , Yoshihara Masaharu , Yasui Wataru , Mukaida Naofumi , Haruma Ken , Chayama Kazuaki
Published: October 2002

Tumor angiogenesis requires the production of angiogenic factors by tumor and stromal cells. Macrophages are key effectors of angiogenesis and reported to contribute to tumor angiogenesis in several carcinomas. To investigate interactions between tumor cells and macrophages in angiogenesis, we examined macrophage infiltration, tumor vascularity and expression of monocyte chemoattractant protein (MCP)‐1, CC chemokine receptor 2 (CCR2) and vascular endothelial growth factor (VEGF) in 57 archival specimens from patients with esophageal dysplasia (n = 9) and squamous cell carcinomas (n = 48). Expression of MCP‐1 mRNA was also examined by reverse transcriptase‐polymerase chain reaction (RT‐PCR) in 7 esophageal carcinoma cell lines and fresh biopsy specimens from 14 patients. The number of infiltrating macrophages correlated closely with expression of VEGF by tumor cells and with neovascularization. Of the 7 cell lines, 4 (TE‐1, 3, 5 and 13) constitutively expressed MCP‐1 mRNA. In 9 (64.3%) of the 14 patients, MCP‐1 mRNA was expressed at high levels in tumor tissues as compared to normal mucosa. MCP‐1 immunoreactivity increased with the depth of tumor invasion (Tis 0%, T1 26.3%, T2, T3 42.1%). Moreover, macrophage and vessel counts were significantly higher in MCP‐1‐positive tumors than in MCP‐1‐negative tumors. Normal and dysplastic esophageal squamous epithelium showed no staining or faint cytoplasmic staining of MCP‐1. Expression of CCR2 immunoreactivity was detected in the cytoplasm of mononuclear cells but not of vascular endothelial cells. These results suggest that interactions between cancer cells and macrophages are important for tumor angiogenesis. MCP‐1 may play a role in progression of human esophageal carcinoma through its role in angiogenesis. © 2002 Wiley‐Liss, Inc.
CD44 stimulation by fragmented hyaluronic acid induces upregulation of urokinase‐type plasminogen activator and its receptor and subsequently facilitates invasion of human chondrosarcoma cells
Authors: Kobayashi Hiroshi , Suzuki Mika , Kanayama Naohiro , Nishida Takashi , Takigawa Masaharu , Terao Toshihiko
Published: October 2002

It has been established that fragmented hyaluronic acid (HA), but not native high molecular weight HA, can induce angiogenesis, cell proliferation and migration. We have studied the outside‐in signal transduction pathways responsible for fragmented HA‐mediated cancer cell invasion. In our study, we have studied the effects of CD44 stimulation by ligation with HA upon the expression of matrix metalloproteinases (MMPs)‐2 and ‐9 as well as urokinase‐type plasminogen activator (uPA), its receptor (uPAR) and its inhibitor (PAI‐1) and the subsequent induction of invasion of human chondrosarcoma cell line HCS‐2/8. Our study indicates that (i) CD44 stimulation by fragmented HA upregulates expression of uPA and uPAR mRNA and protein but does not affect MMPs secretion or PAI‐1 mRNA expression; (ii) the effects of HA fragments are critically HA size dependent: high molecular weight HA is inactive, but lower molecular weight fragmented HA (Mr 3.5 kDa) is active; (iii) cells can bind avidly Mr 3.5 kDa fragmented HA through a CD44 molecule, whereas cells do not effectively bind higher Mr HA; (iv) a fragmented HA induces phosphorylation of MAP kinase proteins (MEK1/2, ERK1/2 and c‐Jun) within 30 min; (v) CD44 is critical for the response (activation of MAP kinase and upregulation of uPA and uPAR expression); and (vi) cell invasion induced by CD44 stimulation with a fragmented HA is inhibited by anti‐CD44 mAb, MAP kinase inhibitors, neutralizing anti‐uPAR pAb, anti‐catalytic anti‐uPA mAb or amiloride. Therefore, our study represents the first report that CD44 stimulation induced by a fragmented HA results in activation of MAP kinase and, subsequently, enhances uPA and uPAR expression and facilitates invasion of human chondrosarcoma cells. © 2002 Wiley‐Liss, Inc.
DCR3 locus is a predictive marker for 5‐fluorouracil‐based adjuvant chemotherapy in colorectal cancer
Authors: Mild Gabriele , Bachmann Felix , Boulay Jean‐Louis , Glatz Katharina , Laffer Urban , Lowy Adam , Metzger Urs , Reuter Jürgen , Terracciano Luigi , Herrmann Richard , Rochlitz Christoph
Published: October 2002

Adjuvant chemotherapy reduces the incidence of distant metastasis and increases survival of patients with colorectal cancer. However, predictive markers are needed to define subsets of patients with stage II and III disease that may benefit from adjuvant treatment. A secreted member of the TNF receptor superfamily, the decoy receptor 3 (DcR3), was reported to be amplified in colorectal cancer as a negative regulator of Fas‐mediated apoptosis. We analyzed DcR3 gene copy number and protein expression in a large series of tumors from a randomized multicenter trial of 5‐fluorouracil/mitomycin C (FU/MMC) adjuvant chemotherapy of the Swiss Group for Clinical Cancer Research (SAKK 40/81), using real‐time quantitative PCR and immunohistochemistry on tumor microarrays. Results of gene status and protein expression of DcR3 were correlated with disease‐free and overall survival of patients. We observed amplification of the DcR3 gene in 185/294 (63%) and overexpression of the DcR3 protein in 163/223 (73%) of colorectal tumors. Multivariate analysis showed no prognostic effect of DcR3 gene amplification and protein overexpression. However, adjuvant chemotherapy was significantly more beneficial in patients with normal DcR3 gene copy number than in patients with amplification (DFS: HR 2.84, 95% CI 1.16–6.98, p = 0.02; OS: HR 3.15, 95% CI 1.19–8.32, p = 0.02), whereas DcR3 protein overexpression did not influence the effect of adjuvant chemotherapy (DFS: HR 1.02, 95% CI 0.65–1.60, p = 0.95; OS: HR 0.95, 95% CI 0.61–1.49, p = 0.83). We conclude that amplification of the 20q13 locus is a predictive marker for adjuvant chemotherapy in colorectal cancer. © 2002 Wiley‐Liss, Inc.
Toward a more complete recognition of immunoreactive antigens in squamous cell lung carcinoma
Authors: Diesinger Isabel , Bauer Christine , Brass Nicole , Schaefers Hans‐Joachim , Comtesse Nicole , Sybrecht Gerhard , Meese Eckart
Published: October 2002

There is very limited knowledge about the antibody response against tumor‐expressed antigens in lung cancer. To arrive at a more complete picture of lung cancer antigens, we generated 2 cDNA libraries from squamous cell lung carcinoma and isolated 15 immunogenic antigens using autologous sera. Among the antigens most frequently identified were the lymphoid blast crisis oncogene (LBC), an unknown hypothetical protein and the p53‐binding protein (TP53 BP), which have already been associated with tumor development. Of the immunogenic antigens, 6 map to chromosomes that are frequently altered in squamous cell lung carcinoma. SEREX database analysis showed that 7 of the identified immunogenic antigens have been associated with the humoral immune response in other human tumors. Screening with heterologous sera of patients with lung carcinoma identified 4 antigens, including human protein kinase C and TP53 BP, which have also been found by autologous screening. Only 1 of the 15 identified antigens reacted with any of the 36 control sera, which were taken from individuals without known disease. Sera from adenocarcinoma and large cell carcinoma of the lung were not reactive for the antigens. In summary, using 2 newly established cDNA libraries, we isolated 15 novel antigens, which were subsequently evaluated for the frequency of their corresponding antibodies in autologous, normal and heterologous sera; their chromosomal localization; and their correlation with survival after surgery. © 2002 Wiley‐Liss, Inc.
Association of chili pepper consumption, low socioeconomic status and longstanding gallstones with gallbladder cancer in a Chilean population
Authors: Serra Iván , Yamamoto Masaharu , Calvo Alfonso , Cavada Gabriel , Báez Sergio , Endoh Kazuo , Watanabe Hidenobu , Tajima Kazuo
Published: October 2002

We explored the risk factors for gallbladder cancer and explanations for its sharp and constant incidence increase in Chile since the 1970s. We compared 114 consecutive patients with verified gallbladder cancer, diagnosed 1992–1995, to 114 matched hospital patients with gallstones, using conditional logistic regression analysis. Low education showed a nonsignificant positive relationship with gallbladder cancer [odds ratio (OR) = 2.3, 95% confidence interval (CI) 0.8–6.2], and low socioeconomic level showed a significant relationship (OR = 5.0, 95% CI 1.5–17.3). A very long history of gallstone disease was significantly more prevalent among cases (OR = 11.0, 95% CI 1.4–85.2). Significant red chili pepper consumption was observed in gallbladder cancer patients (OR = 2.9, 95% CI 1.6–5.2). Low intake of both fresh fruit and sugar as soft drinks was associated with gallbladder cancer, with ORs of 6.4 (95% CI 1.4–30.3) and 3.6 (95% CI 1.3–10.1), respectively. Multivariate analysis kept only a very low socioeconomic status and red chili pepper consumption as significant independent risk factors for gallbladder cancer, ORs of 6.3 (95% CI 1.7–23.0) and 3.2 (95% CI 1.7–5.9). Longstanding gallstone cases were removed from the multivariate model because all were in the low and very low socioeconomic groups, reinforcing the association. Patients with gallbladder cancer differed from matched controls by exhibiting lower socioeconomic levels, having a much longer history of gallstone disease and presenting a dietary pattern characterized by high red chili pepper consumption and low fresh fruit intake. © 2002 Wiley‐Liss, Inc.
Oxidative DNA damage by a common metabolite of carcinogenic nitrofluorene andN‐acetylaminofluorene
Authors: Murata Mariko , Yoshiki Yumiko , Tada Mariko , Kawanishi Shosuke
Published: October 2002

Both carcinogenic NF and AAF are metabolized to a commonN‐hydroxy metabolite, N‐OH‐AF. We investigated oxidative DNA damage by N‐OH‐AF, using32P‐labeled human DNA fragments from the humanp53andp16tumor‐suppressor genes and the c‐Ha‐ras‐1 protooncogene. N‐OH‐AF caused Cu(II)‐mediated DNA damage, and endogenous reductant NADH markedly enhanced this process. Catalase and bathocuproine, a Cu(I)‐specific chelator, decreased the DNA damage, suggesting the involvement of H2O2and Cu(I). N‐OH‐AF induced piperidine‐labile lesions frequently at thymine and cytosine residues. With formamidopyrimidine‐DNA glycosylase treatment, N‐OH‐AF induced cleavage at guanine residues, especially of the ACG sequence complementary to codon 273, a well‐known hot spot of thep53gene. N‐OH‐AF dose‐dependently induced 8‐oxodG formation in the presence of Cu(II) and NADH. Treatment with N‐OH‐AF increased amounts of 8‐oxodG in HL‐60 cells compared to the H2O2‐resistant clone HP100, supporting the involvement of H2O2. The present study demonstrates that theN‐hydroxy metabolite of NF and AAF induces oxidative DNA damage through H2O2in both a cell‐free system and cultured human cells. We conclude that oxidative DNA damage may play an important role in the carcinogenic process of NF and AAF in addition to previously reported DNA adduct formation. © 2002 Wiley‐Liss, Inc.
Cytoplasmic and/or nuclear accumulation of the β‐catenin protein is a frequent event in human osteosarcoma
Authors: Haydon Rex C. , Deyrup Andrea , Ishikawa Akira , Heck Robert , Jiang Wei , Zhou Lan , Feng Tao , King David , Cheng Hongwei , Breyer Benjamin , Peabody Terrance , Simon Michael A. , Montag Anthony G. , He Tong‐Chuan
Published: October 2002

The molecular events that precede the development of osteosarcoma, the most common primary malignancy of bone, are unclear, and concurrent molecular and genetic alterations associated with its pathogenesis have yet to be identified. Recent studies suggest that activation of β‐catenin signaling may play an important role in human tumorigenesis. To investigate the potential role of β‐catenin deregulation in human osteosarcoma, we analyzed a panel of 47 osteosarcoma samples for β‐catenin accumulation using immunohistochemistry. Potential activating mutations were investigated by sequencing exon 3 of the β‐catenin gene in genomic DNA isolated from tumor samples. Our findings revealed cytoplasmic and/or nuclear accumulation of β‐catenin in 33 of 47 samples (70.2%); however, mutation analysis failed to detect any genetic alterations within exon 3, suggesting that other regulatory mechanisms may play an important role in activating β‐catenin signaling in osteosarcoma. In our survival analysis, β‐catenin deregulation conferred a hazard ratio of 1.05, indicating that β‐catenin accumulation does not appear to be of prognostic value for osteosarcoma patients. When analyzed against other clinicopathologic parameters, β‐catenin accumulation correlated only with younger age at presentation (26.4 vs. 39.8 years). Nevertheless, our results demonstrate that the deregulation of β‐catenin signaling is a common occurrence in osteosarcoma that is implicated in the pathogenesis of osteosarcoma. © 2002 Wiley‐Liss, Inc.
Continuing rising trend in oesophageal adenocarcinoma
Authors: Powell Jean , McConkey Chris C. , Gillison E. Walford , Spychal Robert T.
Published: October 2002

Our study provides an update of the incidence of oesophageal cancer in the West Midland region of England and Wales from 1992–96. A total of 2,671 cases of oesophageal cancer were identified during the 5‐year study period, with an age‐standardised annual incidence (ASR) of 5.24 per 100,000 (95% CI: 5.02, 5.45). Similar numbers of adenocarcinoma and squamous cell carcinoma were found. Only 152 (5.6%) had no histology. There was a 5‐fold difference in age‐standardised annual incidence rates between males and females for adenocarcinoma of oesophagus, but no gender difference for squamous cell carcinoma. The parallel but higher ASR in males compared to females for adenocarcinoma of both oesophagus and cardia merits further investigation. The similarities in the patterns of age‐ and sex‐specific rates and in the socioeconomic profiles could indicate a common aetiology for adenocarcinoma of oesophagus and gastric cardia. Quality control in Cancer Registries needs to focus on the accuracy and consistency of subsite classification to ensure that trends in incidence are identified. In the absence of accurate subsite classification of stomach cancers, the proportions of adenocarcinoma and squamous cell carcinoma of oesophagus (or the absolute rate of adenocarcinoma of oesophagus) may provide a useful tool in indicating whether adenocarcinoma of gastric cardia is likely to be increasing in incidence. © 2002 Wiley‐Liss, Inc.
Microsatellite instability in thorotrast‐induced human intrahepatic cholangiocarcinoma
Authors: Liu Duo , Momoi Hirohito , Li Li , Ishikawa Yuichi , Fukumoto Manabu
Published: October 2002

Thorotrast, a colloidal suspension of radioactive 232ThO2 that emits α particles, was used as a radiographic contrast during World War II. It is known to induce liver cancers, most frequently ICC, decades after injection. Since radiation induces genomic instability, we analyzed MSI in Thorotrast‐induced ICC. The frequency of MSI+ cases was 62.5% in Thorotrast ICC, whereas it was 22.7% in non‐Thorotrast ICC. However, frameshift mutations of mononucleotide repeats were not observed in Thorotrast ICC. In addition, the MSI+ phenotype was not associated with the quantity of Thorotrast deposited or the latency period of ICC induction. Promoter regions of both the hMLH1 and the hMSH2 MMR genes tended to be hypermethylated in the tumor part compared to the adjacent nontumor part in Thorotrast ICC. Methylation of the hMLH1 promoter was associated with the MSI+ phenotype in Thorotrast ICC. In contrast, methylation status of these promoter regions was not related to MSI in non‐Thorotrast ICC cases. These findings suggest that MSI induced by exposure to Thorotrast mainly reflects clonal expansion of cancer cells and is partly due to inactivation of hMLH1 by hypermethylation. © 2002 Wiley‐Liss, Inc.
Both the epitope specificity and isotype are important in the antitumor effect of monoclonal antibodies against Her‐2/neu antigen
Authors: Kim Kwang‐Mi , Shin Eun‐Young , Moon Jeoung‐Hyun , Heo Tae‐Hwe , Lee Joon Youb , Chung Yeonseok , Lee Yoon‐Jung , Cho Hyun‐Mi , Shin Seung‐Uon , Kang Chang‐Yuil
Published: October 2002

The Her‐2/neu oncogene, which encodes a growth factor receptor, was implicated in the malignancy of human adenocarcinomas. Antibodies directed to this molecule have been previously shown to have an antitumor effect in vivo. In an attempt to understand the mechanisms of the antitumor activity, we generated 2 monoclonal antibodies (mAbs), HRO G1 and HRT G1, that recognize different epitopes on Her‐2/neu. Both of the mAbs bound HER2/neu on the tumor surface, resulting in phosphorylation of HER2/neu. We also generated IgG2a and IgG2b mAbs from these 2 mAbs, respectively. The results of in vitro studies showed that these anti‐Her‐2/neu mAbs could not inhibit the growth of the tumor cells that express Her‐2/neu molecules by themselves. However, in an antibody‐dependent cellular cytotoxicity study using mouse splenocytes as effector cells, HRT mAbs had antitumor activities superior to those of HRO mAbs, indicating that the epitope specificity may also partake in antibody‐dependent cellular cytotoxicity with antibody isotype. In a complement‐dependent cytotoxicity study, the IgG2a and IgG2b mAbs showed stronger effects than IgG1 isotype mAbs irrespective of the epitope specificities. The results of in vivo studies also showed that HRT mAbs had superior antitumor activity to those of HRO mAbs. The antitumor activity was most prominent in the HRT G2b isotype among HRT mAbs. HRT G1 also showed a moderate antitumor effect, while HRT G2a showed only slight inhibition effect. These data indicate that both the epitope specificity and the differences in Fc region of mAbs could play important roles in the antitumor activities. © 2002 Wiley‐Liss, Inc.
Estrogen receptor α expression in normal human breast epithelium is consistent over time
Authors: Khan Seema A. , Yee Kimberly A. , Kaplan Cassandra , Siddiqui Josephine F.
Published: October 2002

If increased expression of estrogen receptor α (ER) in benign breast epithelium increases susceptibility to breast cancer, such overexpression should be stable over time. There are no published data regarding this important aspect of ER expression in breast epithelium. We examined the temporal consistency of ER expression in the normal breast tissue of 28 women who had 2 separate breast surgical procedures, at least 6 months apart (mean interval, 2.8 years). Paraffin embedded breast tissue blocks containing an adequate sample of normal breast epithelium and no cancer, were sectioned and processed using the 6F11 antibody and standard immunohistochemical techniques. The ER labelling index (ER LI) was calculated by counting a mean of 2,000 epithelial cells. The median ER LI at first sampling was 13.6 and at second sample 15.5, with R2 = 0.34 and p = 0.001. The ER LI was categorized into high and low values, using a threshold of 10. Twenty‐four women (85.7%) showed concordance of high and low expression between the 2 samples (p = 0.002). There were 11 women who were premenopausal at both time points. Among them, much of the variation in ER LI was explained by differences in the menstrual cycle day at the time of sampling and adding the day of cycle to the regression model substantially improved the correlation between first and second labeling indices. These data suggest that ER expression of normal breast tissue is fairly consistent over time and support the notion that overexpression of ER in normal epithelium is a constant feature of the high risk breast. © 2002 Wiley‐Liss, Inc.
Submicromolar doses of alkyl‐lysophospholipids induce rapid internalization, but not activation, of epidermal growth factor receptor and concomitant MAPK/ERK activation in A431 cells
Authors: Ruiter Gerald A. , Verheij Marcel , Zerp Shuraila F. , Moolenaar Wouter H. , Van Blitterswijk Wim J.
Published: October 2002

Synthetic ALPs, e.g., Et‐18‐OCH3 and HePC, are anticancer agents that accumulate in cell membranes, where they interfere with lipid‐mediated signal transduction. We previously reported that ALPs, when added at micromolar concentrations (5–25 μM), inhibit growth factor–induced MAPK/ERK activation and enhance radiation‐induced apoptosis. We now show that, at nanomolar doses (10–500 nM), ALPs activate the MAPK/ERK pathway in A431 cells without stimulating cell proliferation. Strikingly, ALPs (500 nM) also trigger rapid clustering and internalization of the EGFR in A431 cells. Tyrphostin AG1478, an EGFR tyrosine kinase inhibitor, blocks ALP‐induced MAPK/ERK activation but not EGFR internalization. We found no evidence for ALPs acting via G protein–coupled receptors and/or transactivation of EGFRs, as determined by calcium mobilization, EGFR phosphorylation and Grb2 binding assays. Since ALPs readily intercalate into the plasma membrane, our data suggest that they induce subtle changes in the lipid microenvironment of the EGFR, resulting in clustering and internalization of the EGFR and concomitant MAPK/ERK activation. © 2002 Wiley‐Liss, Inc.
In vivo angiogenic activity of neuroblastoma correlates with MYCN oncogene overexpression
Authors: Ribatti Domenico , Raffaghello Lizzia , Pastorino Fabio , Nico Beatrice , Brignole Chiara , Vacca Angelo , Ponzoni Mirco
Published: October 2002

Neuroblastoma (NB) is the most common malignant solid tumor in early childhood. Amplification of the MYCN oncogene is associated with a more malignant course of disease and poor outcome. The role that MYCN plays in the regulation of angiogenesis in NB remains unclear. To better elucidate this matter, fresh biopsy samples from 21 patients, 10 with MYCN‐amplified tumors (defined as having >10 copies of the oncogene) and 11 with nonamplified tumors, were tested for their angiogenic capacity using the chick embryo chorioallantoic membrane assay, a useful model for such investigation. Moreover, using the same experimental model, conditioned media obtained from 5 different human NB cell lines MYCN‐amplified (HTLA‐230, LAN‐5 and GI‐LI‐N) or nonamplified (ACN and SH‐SY5Y) and biopsy fragments obtained from xenografts derived from 4 NB cell lines (HTLA‐230, GI‐LI‐N, ACN and SH‐SY5Y) injected in nude mice were assayed for angiogenic potential. Our results clearly demonstrated that MYCN amplification parallels angiogenesis in NB. When fresh biopsy samples from patients, CM derived from NB cell lines and biopsy fragments derived from xenografts of the same cell lines injected in nude mice were tested, the response was univocal: the angiogenic response, evaluated both macroscopically and microscopically, was significantly higher in the MYCN‐amplified specimens compared to the nonamplified ones. © 2002 Wiley‐Liss, Inc.
Consultants warn about worrying treasury risks
Authors: Gregg Cathryn R. , Gallanis Michael A.
Published: October 2002

The authors, who are professional treasury consultants, sound a warning about three troubling risks: IT fraud,
payment fraud, and business interruption. Are you ready to handle them? © 2002 Wiley
Periodicals, Inc.
Can the CFO stop white collar crime?
Authors: Christensen Jo Ann , Byington J. Ralph
Published: October 2002

When there's an accounting scandal, one of the first casualties can be the CFO. What can the CFO do to protect himself—and his company—from white collar crime, as the number of scandals continues to grow?© 2002 Wiley Periodicals, Inc.
CRM: The customer isn't always right
Authors: Bergeron Bryan
Published: October 2002

Service companies believe two myths, says the author. One is that the customer is always right. And the second
is that anyone willing to part with his money should be treated as a valid customer. Instead, the author argues,
successful companies target customers who fit their business model. They're embracing a process called
Customer Relationship Management (CRM). © 2002 Wiley Periodicals, Inc.
Evidence on the operation of ATP‐induced capacitative calcium entry in breast cancer cells and its blockade by 17β‐estradiol
Authors: Rossi Ana M. , Picotto Gabriela , de Boland Ana R. , Boland Ricardo L.
Published: October 2002

Little is known about the regulation of cytosolic calcium Ca2+ levels ([Ca2+]i) in breast cancer cells. We investigated the existence of capacitative calcium entry (CCE) in the tumorigenic cell line MCF‐7 and its responsiveness to ATP. MCF‐7 cells express purinergic receptors as well as estrogen receptors (ER). Depletion of calcium stores with thapsigargin (TG, 500 nM) or ATP (10 μM) in the absence of extracellular Ca2+, resulted in a rapid and transient elevation in [Ca2+]i. After recovery of basal levels, Ca2+ readmission (1.5 mM) to the medium increased Ca2+ influx (twofold over basal), reflecting pre‐activation of a CCE pathway. Cells pretreated with TG were unable to respond to ATP, thus indicating that the same Ca2+ store is involved in their response. Moreover, IP3‐dependent ATP‐induced calcium mobilization and CCE were completely blocked using compound U‐73122, an inhibitor of phospholipase C. Compound 2‐APB (75 μM) and Gd3+ (10 μM), antagonists of the CCE pathway, completely prevented ATP‐stimulated capacitative Ca2+ entry. CCE in MCF‐7 cells was highly permeable to Mn2+ and to the Ca2+ surrogate Sr2+. Mn2+ entry sensitivity to Gd3+ matched that of the Ca2+ entry pathway. 17β‐estradiol blocked ATP‐induced CCE, but was without effect on TG‐induced CCE. Besides, the estrogen blockade of the ATP‐induced CCE was completely abolished by preincubation of the cells with an ER monoclonal antibody. ER α immunoreactivity could also be detected in a purified plasma membrane fraction of MCF‐7 cells. These results represent the first evidence on the operation of a ATP‐responsive CCE pathway in MCF‐7 cells and also indicate that 17β‐estradiol interferes with this mechanism by acting at the cell surface level. J. Cell. Biochem. 87: 324–333, 2002. © 2002 Wiley‐Liss, Inc.