bluuebunny/crossref_metadata_2025 huggingface dataset:
Mutations in the Focal Adhesion Targeting Region of Deleted in Liver Cancer-1 Attenuate Their Expression and Function
Authors: Liao Yi-Chun , Shih Yi-Ping , Lo Su Hao
Published: October 2008

Deleted in liver cancer-1 (DLC-1) is a RhoGTPase-activating protein (RhoGAP) domain containing tumor suppressor that is often down-regulated in various cancer types. Previously, we have shown that DLC-1 is recruited to focal adhesions by binding to the Src homology 2 domains of tensins and the focal adhesion localization is critical for the tumor suppression activity of DLC-1. To investigate whether mutations in the focal adhesion targeting (FAT) region might occur and attenuate the expression, localization, and function of DLC-1, we have first mapped the FAT region to the amino acid residues from 201 to 500, and then sequenced cDNAs and genomic DNAs encoding the FAT region from cancer patients. Several missense and nonsense mutations were detected. All missense mutations were further examined for the potential effect on the function of DLC-1. Although these mutations did not seem to affect the focal adhesion localization of DLC-1, the activities of suppressing tumor cell growth were impaired in two mutants: T301K and S308I. Consistent with the fact that the RhoGAP activity of DLC-1 is essential for inhibiting tumor cell growth, the RhoGAP activities were significantly reduced in these mutants, suggesting that the FAT region also contains a regulatory element for its COOH-terminal RhoGAP domain. Our studies have shown that mutations in DLC-1 may lead to loss of function and contribute to the tumorigenesis, and have revealed an allosteric regulation site for its RhoGAP activity. [Cancer Res 2008;68(19):7718–22]
Androgenic Control of Transforming Growth Factor-β Signaling in Prostate Epithelial Cells through Transcriptional Suppression of Transforming Growth Factor-β Receptor II
Authors: Song Kyung , Wang Hui , Krebs Tracy L. , Kim Seong-Jin , Danielpour David
Published: October 2008

The androgen receptor cross-talks with transforming growth factor-β (TGF-β) through mechanisms that remain poorly understood. Here we provide strong evidence that 5α-dihydrotestosterone (DHT) intercepts the ability of prostate epithelial cells to undergo TGF-β–induced apoptosis, and present a new model for this androgenic effect. We report that DHT decreases the level of TGF-β receptor II (TβRII) through a transcriptional mechanism, leading to suppression of the ability of TGF-β to down-regulate expression of Bcl-xL and cyclin Ds, activate caspase-3, and induce apoptosis. Promoter analysis, DNA pulldown, and electrophoretic mobility shift assays support that transcriptional down-regulation of TβRII by DHT occurs through Sp1/Sp3 response elements, with the binding of Sp1 to the TβRII promoter being suppressed by DHT, largely driven by loss of Sp1 protein and/or activity. These results provide fresh insight on the mechanism of growth control by androgens and the progression of prostate cancer to androgen independence. [Cancer Res 2008;68(19):8173–82]
Identification of Transformation-Related Pathways in a Breast Epithelial Cell Model Using a Ribonomics Approach
Authors: Mazan-Mamczarz Krystyna , Hagner Patrick R. , Dai Bojie , Wood William H. , Zhang Yongqing , Becker Kevin G. , Liu Zhenqui , Gartenhaus Ronald B.
Published: October 2008

The aberrant expression of many genes is a common feature in the malignant transformation of cells. In mammalian cells, posttranscriptional gene regulatory processes are emerging as critical determinants controlling gene expression both in physiologic and pathologic conditions. These regulatory mechanisms are directed primarily by the interaction of mRNAs with specific RNA-binding proteins (RBP). There is an emerging body of data demonstrating that two RBPs, AUF1 and HuR, can antagonistically affect the posttranscriptional fate of target mRNAs, as well as concurrently bind to common target transcripts. Employing MCT-1 oncogene-mediated transformation of immortalized breast epithelial MCF10A cells, we characterized the largely reciprocal association of these two RBPs with target mRNAs and their influence on protein expression vis-a-vis cellular transformation. Using a ribonomics approach, we identified mRNAs from cancer-related pathways whose association with AUF1 and/or HuR were altered when comparing immortalized with transformed MCF10A cells. Significantly, we were able to show that knockdown of HuR expression using RNA interference reduced anchorage-independent growth capacity in transformed MCF10A cells and decreased protein expression of a number of validated target genes. Our data show that the global alterations in binding of HuR and AUF1 with target transcripts have a critical role in posttranscriptional regulation of genes encoding proteins involved in breast epithelial cell transformation. These findings further support the feasibility of using a ribonomics approach for the identification of cancer-related pathways. [Cancer Res 2008;68(19):7730–5]
Identification of GRP75 as an Independent Favorable Prognostic Marker of Neuroblastoma by a Proteomics Analysis
Authors: Hsu Wen-Ming , Lee Hsinyu , Juan Hsueh-Fen , Shih Yu-Yin , Wang Bo-Jeng , Pan Chien-Yuan , Jeng Yung-Ming , Chang Hsiu-Hao , Lu Meng-Yao , Lin Kai-Hsin , Lai Hong-Shiee , Chen Wei-Jao , Tsay Yeou-Guang , Liao Yung-Feng , Hsieh Fon-Jou
Published: October 2008

Purpose: Neuroblastoma (NB) is a heterogeneous neoplasm. Detailed biological discrimination is critical for the effective treatment of this disease. Because the tumor behavior of NB is closely associated with the histologic state of differentiation, we thus aimed to identify novel differentiation-associated markers of NB with prognostic implication.

Experimental Design: A human NB cell line SH-SY5Y was used as a model system to explore potential biomarkers for the differentiation of NB by proteomic analyses. Seventy-two NB tumor tissues were subsequently investigated by immunohistochemistry to validate the correlations between the expression of a novel prognostic marker, various clinicopathologic and biological factors, and patient survival.

Results: Using two-dimensional differential gel electrophoresis, we found a total of 24 spots of proteins in SH-SY5Y cells whose expression was enhanced following differentiation. Glucose-regulated protein 75 (GRP75) was unambiguously identified as one of the five proteins that were dramatically up-regulated following differentiation. Immunohistochemical analyses of 72 NB tumor tissues further revealed that positive GRP75 immunostaining is strongly correlated with differentiated histologies (P < 0.001), mass-screened tumors (P = 0.016), and early clinical stages (P < 0.001) but inversely correlated with MYCN amplification (P = 0.010). Univariate and multivariate survival analyses showed that GRP75 expression is an independent favorable prognostic factor.

Conclusions: The present findings clearly showed that our proteomics-based novel experimental paradigm could be a powerful tool to uncover novel biomarkers associated with the differentiation of NB. Our data also substantiate an essential role of GRP75 in the differentiation of NB.
The Use of Genomics in Clinical Trial Design
Authors: Simon Richard
Published: October 2008

Many cancer treatments benefit only a minority of patients who receive them. This results in an enormous burden on patients and on the health care system. The problem will become even greater with the increasing use of molecularly targeted agents whose benefits are likely to be more selective unless the drug development process is modified to include codevelopment of companion diagnostics. Whole genome biotechnology and decreasing costs of genome sequencing make it increasingly possible to achieve an era of predictive medicine in oncology therapeutics. The challenges are numerous and substantial but are not primarily technological. They involve organizing publicly funded diagnostics of deregulated pathways, adopting new paradigms for drug development, and developing incentives for industry to incur the complexity and expense of codevelopment of drugs and companion diagnostics. This article reviews some designs for phase III clinical trials that may facilitate movement to a more predictive oncology.
Validation of Analytic Methods for Biomarkers Used in Drug Development
Authors: Chau Cindy H. , Rixe Olivier , McLeod Howard , Figg William D.
Published: October 2008

The role of biomarkers in drug discovery and development has gained precedence over the years. As biomarkers become integrated into drug development and clinical trials, quality assurance and, in particular, assay validation become essential with the need to establish standardized guidelines for analytic methods used in biomarker measurements. New biomarkers can revolutionize both the development and use of therapeutics but are contingent on the establishment of a concrete validation process that addresses technology integration and method validation as well as regulatory pathways for efficient biomarker development. This perspective focuses on the general principles of the biomarker validation process with an emphasis on assay validation and the collaborative efforts undertaken by various sectors to promote the standardization of this procedure for efficient biomarker development.
Salivary Proteomics for Oral Cancer Biomarker Discovery
Authors: Hu Shen , Arellano Martha , Boontheung Pinmanee , Wang Jianghua , Zhou Hui , Jiang Jiang , Elashoff David , Wei Roger , Loo Joseph A. , Wong David T.
Published: October 2008

Purpose: This study aims to explore the presence of informative protein biomarkers in the human saliva proteome and to evaluate their potential for detection of oral squamous cell carcinoma (OSCC).

Experimental Design: Whole saliva samples were collected from patients (n = 64) with OSCC and matched healthy subjects (n = 64). The proteins in pooled whole saliva samples of patients with OSCC (n = 16) and matched healthy subjects (n = 16) were profiled using shotgun proteomics based on C4 reversed-phase liquid chromatography for prefractionation, capillary reversed-phase liquid chromatography with quadruple time-of-flight mass spectrometry, and Mascot sequence database searching. Immunoassays were used for validation of the candidate biomarkers on a new group of OSCC (n = 48) and matched healthy subjects (n = 48). Receiver operating characteristic analysis was exploited to evaluate the diagnostic value of discovered candidate biomarkers for OSCC.

Results: Subtractive proteomics revealed several salivary proteins at differential levels between the OSCC patients and matched control subjects. Five candidate biomarkers were successfully validated using immunoassays on an independent set of OSCC patients and matched healthy subjects. The combination of these candidate biomarkers yielded a receiver operating characteristic value of 93%, sensitivity of 90%, and specificity of 83% in detecting OSCC.

Conclusion: Patient-based saliva proteomics is a promising approach to searching for OSCC biomarkers. The discovery of these new targets may lead to a simple clinical tool for the noninvasive diagnosis of oral cancer. Long-term longitudinal studies with large populations of individuals with oral cancer and those who are at high risk of developing oral cancer are needed to validate these potential biomarkers.
Liposome-Encapsulated Curcumin Suppresses Growth of Head and Neck Squamous Cell CarcinomaIn vitroand in Xenografts through the Inhibition of Nuclear Factor κB by an AKT-Independent Pathway
Authors: Wang Dorothy , Veena Mysore S. , Stevenson Kerry , Tang Christopher , Ho Baran , Suh Jeffrey D. , Duarte Victor M. , Faull Kym F. , Mehta Kapil , Srivatsan Eri S. , Wang Marilene B.
Published: October 2008

Purpose: The purpose of this study was to determine whether a liposomal formulation of curcumin would suppress the growth of head and neck squamous cell carcinoma (HNSCC) cell lines CAL27 and UM-SCC1 in vitro and in vivo.

Experimental Design: HNSCC cell lines were treated with liposomal curcumin at different doses and assayed for in vitro growth suppression using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. A reporter gene assay was done on cell lines to study the effect of liposomal curcumin on nuclear factor κB (NFκB) activation. Western blot analysis was done to determine the effect of curcumin on the expression of NFκB, phospho-IκBα, phospho-AKT (pAKT), phospho-S6 kinase, cyclin D1, cyclooxygenase-2, matrix metalloproteinase-9, Bcl-2, Bcl-xL, Mcl-1L, and Mcl-1S. Xenograft mouse tumors were grown and treated with intravenous liposomal curcumin. After 5 weeks, tumors were harvested and weighed. Immunohistochemistry and Western blot analyses were used to study the effect of liposomal curcumin on the expression of NFκB and pAKT.

Results: The addition of liposomal curcumin resulted in a dose-dependent growth suppression of both cell lines. Liposomal curcumin treatment suppressed the activation of NFκB without affecting the expression of pAKT or its downstream target phospho-S6 kinase. Expression of cyclin D1, cyclooxygenase-2, matrix metalloproteinase-9, Bcl-2, Bcl-xL, Mcl-1L, and Mcl-1S were reduced, indicating the effect of curcumin on the NFκB pathway. Nude mice xenograft tumors were suppressed after 3.5 weeks of treatment with i.v. liposomal curcumin, and there was no demonstrable toxicity of liposomal curcumin upon autopsy. Immunohistochemistry and Western blot analysis on xenograft tumors showed the inhibition of NFκB without affecting the expression of pAKT.

Conclusions: Liposomal curcumin suppresses HNSCC growth in vitro and in vivo. The results suggest that liposomal curcumin is a viable nontoxic therapeutic agent for HNSCC that may work via an AKT-independent pathway.
Transesophageal Endoscopic Ultrasound with Fine Needle Aspiration in the Preoperative Staging of Malignant Pleural Mesothelioma
Authors: Tournoy Kurt G. , Burgers Sjaak A. , Annema Jouke T. , Vermassen Frank , Praet Marleen , Smits Marianne , Klomp Houke M. , van Meerbeeck Jan P. , Baas Paul
Published: October 2008

Purpose: Surgical resection as part of a multimodality approach in malignant pleural mesothelioma (MPM) has a high morbidity and mortality. Because mediastinal lymph node (MLN) metastases are a negative prognostic factor, preoperative staging is of paramount importance. Transesophageal endoscopic ultrasound with real-time guided fine needle aspiration (EUS-FNA) enables accurate MLN staging in lung cancer.

Experimental Design: The feasibility and yield of EUS-FNA in MLN staging were prospectively analyzed in patients with presumed early-stage MPM considered for multimodality therapy. MLN reference pathology was defined by either pathologic staging or the formal demonstration of malignant cells by either EUS-FNA or mediastinoscopy.

Results: Thirty-two consecutive patients (81% males; median age, 61 years) with proven MPM underwent EUS-FNA. In 11 (34%) patients, a negative EUS-FNA or mediastinoscopy was not confirmed by surgical MLN dissection because of clinical deterioration or disease progression. In 21 (66%) patients, a formal pathology of the MLN was obtained and staging with EUS-FNA was positive in 4 (19%). Mediastinoscopy did not result in a greater yield of MLN metastasis as compared with EUS-FNA. Thoracotomy with complete lymph node dissection was done in 17 (81%). The overall prevalence of MLN metastasis was 24%, and the sensitivity of EUS-FNA was 80% (95% confidence interval, 28-99%) with a specificity of 100% (95% confidence interval, 79-100%). One patient had esophageal perforation related to EUS-FNA.

Conclusions: EUS-FNA is feasible and sensitive for MLN staging in patients with MPM who are candidate for multimodality treatment. These data warrant further evaluation.
Single-Agent and Combination Therapeutic Strategies to Inhibit Hepatocyte Growth Factor/MET Signaling in Cancer
Authors: Toschi Luca , Jänne Pasi A.
Published: October 2008

Receptor tyrosine kinases are often aberrantly activated in human malignancies and contribute to cancer development and progression. Specific receptor tyrosine kinase inhibitors have been shown to be clinically effective therapies in subsets of cancer patients with either hematologic or solid tumors. Activation of the hepatocyte growth factor (HGF)/MET signaling pathway has been found to play a critical role in oncogenesis, cancer metastasis, and drug resistance. These observations have led to the development of agents that can effectively inhibit HGF/MET signaling through direct inhibition of the receptor (anti-MET antibodies), through inactivation of its ligand HGF (AMG102, L2G7), by interfering with HGF binding to MET (NK4), or by inhibiting MET kinase activity (PHA-665752 and SU11274). Moreover, the combination of anti-MET therapeutic agents with either signal transduction inhibitors (ERBB family or mTOR inhibitors) or with cytotoxic chemotherapy has been evaluated in preclinical models. These studies provide insight into the rational development of combination therapeutic strategies that can be evaluated in clinical trials. This review will discuss different strategies of MET inhibition with a specific focus on combination therapeutic approaches.
Negative Regulation of the Osteoblast Function in Multiple Myeloma through the Repressor Gene E4BP4 Activated by Malignant Plasma Cells
Authors: Silvestris Franco , Cafforio Paola , De Matteo Monica , Calvani Nicola , Frassanito Maria A. , Dammacco Franco
Published: October 2008

Purpose: To explore the pathogenetic mechanisms that suppress the osteoblast function in multiple myeloma because osteogenesis results in defective new bone formation and repair.

Experimental Design: Microarray gene analysis revealed the overexpression of E4BP4, a transcriptional repressor gene, in normal osteoblasts cocultured with myeloma cells that were releasing the parathyroid hormone-related protein (PTHrP). Thus, the effect of E4BP4 was assessed in PTHrP-stimulated osteoblasts by measuring the RNA levels of both Runx2 and Osterix as major osteoblast transcriptional activators. Because E4BP4 is a negative regulator of the cyclooxygenase-2 (COX-2) pathway that drives the expression of both Runx2 and Osterix, these factors were investigated after prostaglandin E2 treatment to overcome the COX-2 defect as well as in E4BP4-silenced osteoblasts. Finally, E4BP4, PTHrP, Osterix, and osteocalcin levels were measured in vivo in patients with bone disease together with the E4BP4 protein in bone biopsies.

Results: E4BP4 was specifically induced by PTHrP and inhibited both Runx2 and Osterix, whereas E4BP4-silenced osteoblasts expressed functional levels of both factors. The prostaglandin E2 treatment of E4BP4-up-regulated osteoblasts promptly restored Runx2 and Osterix activities, suggesting that integrity of COX-2 pathway is essential for their transcription. Down-regulation of Osterix by E4BP4 was confirmed in vivo by its inverse levels in osteoblasts from myeloma patients with increased serum PTHrP, whose bone biopsies expressed the E4BP4 protein.

Conclusions: Our data support the role of E4BP4 as osteoblast transcriptional repressor in inhibiting both Runx2 and Osterix in myeloma bone disease and correlate its effect with the increased PTHrP activity.
Sonic Hedgehog Promotes Desmoplasia in Pancreatic Cancer
Authors: Bailey Jennifer M. , Swanson Benjamin J. , Hamada Tomofumi , Eggers John P. , Singh Pankaj K. , Caffery Thomas , Ouellette Michel M. , Hollingsworth Michael A.
Published: October 2008

Purpose: We investigated the contribution of Sonic hedgehog (SHH) to pancreatic cancer progression.

Experimental Design: We expressed SHH in a transformed primary ductal-derived epithelial cell line from the human pancreas, transformed hTert-HPNE (T-HPNE), and evaluated the effects on tumor growth. We also directly inhibited the activity of SHH in vivo by administering a blocking antibody to mice challenged orthotopically with the Capan-2 pancreatic cancer cell line, which is known to express SHH and form moderately differentiated tumors in nude mice.

Results: Our data provide evidence that expression of SHH influences tumor growth by contributing to the formation of desmoplasia in pancreatic cancer. We further show that SHH affects the differentiation and motility of human pancreatic stellate cells and fibroblasts.

Conclusions: These data suggest that SHH contributes to the formation of desmoplasia in pancreatic cancer, an important component of the tumor microenvironment.
Deletions of CDKN2C in Multiple Myeloma: Biological and Clinical Implications
Authors: Leone Paola E. , Walker Brian A. , Jenner Matthew W. , Chiecchio Laura , Dagrada GianPaolo , Protheroe Rebecca K.M. , Johnson David C. , Dickens Nicholas J. , Brito Jose Luis , Else Monica , Gonzalez David , Ross Fiona M. , Chen-Kiang Selina , Davies Faith E. , Morgan Gareth J.
Published: October 2008

Purpose: Deletions of chromosome 1 have been described in 7% to 40% of cases of myeloma with inconsistent clinical consequences. CDKN2C at 1p32.3 has been identified in myeloma cell lines as the potential target of the deletion. We tested the clinical impact of 1p deletion and used high-resolution techniques to define the role of CDKN2C in primary patient material.

Experimental Design: We analyzed 515 cases of monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), and newly diagnosed multiple myeloma using fluorescence in situ hybridization (FISH) for deletions of CDKN2C. In 78 myeloma cases, we carried out Affymetrix single nucleotide polymorphism mapping and U133 Plus 2.0 expression arrays. In addition, we did mutation, methylation, and Western blotting analysis.

Results: By FISH we identified deletion of 1p32.3 (CDKN2C) in 3 of 66 MGUS (4.5%), 4 of 39 SMM (10.3%), and 55 of 369 multiple myeloma cases (15%). We examined the impact of copy number change at CDKN2C on overall survival (OS), and found that the cases with either hemizygous or homozygous deletion of CDKN2C had a worse OS compared with cases that were intact at this region (22 months versus 38 months; P = 0.003). Using gene mapping we identified three homozygous deletions at 1p32.3, containing CDKN2C, all of which lacked expression of CDKN2C. Cases with homozygous deletions of CDKN2C were the most proliferative myelomas, defined by an expression-based proliferation index, consistent with its biological function as a cyclin-dependent kinase inhibitor.

Conclusions: Our results suggest that deletions of CDKN2C are important in the progression and clinical outcome of myeloma.
Characteristics of Dasatinib- and Imatinib-Resistant Chronic Myelogenous Leukemia Cells
Authors: Okabe Seiichi , Tauchi Tetsuzo , Ohyashiki Kazuma
Published: October 2008

Purpose: Although dual src-family kinase/BCR/ABL inhibitor, dasatinib (BMS-354825), provides therapeutic advantages to imatinib-resistant cells, the mechanism of dasatinib resistance was not fully known.

Experimental Design: We used TF-1 BCR/ABL cells, by introducing the BCR/ABL gene into a leukemia cell line, TF-1 and K562, and established dasatinib- (BMS-R) and imatinib-resistant (IM-R) cells. We characterized chronic myelogenous leukemia drug-resistant cells and examined intracellular signaling.

Results: The IC50 of dasatinib was 0.75 nmol/L (TF-1 BCR/ABL), 1 nmol/L (K562), 7.5 nmol/L (TF-1 BCR/ABL IM-R), 10 nmol/L (K562 IM-R), 15 μmol/L (TF-1 BCR/ABL BMS-R), and 25 μmol/L (K562 BMS-R). The number of BCR/ABL copies in resistant cell lines was the same as the parental cell line by fluorescence in situ hybridization analysis. There was no mutation in Abl kinase. We found that protein levels of BCR/ABL were reduced in dasatinib-resistant cell lines. BCR/ABL protein was increased by treatment of an ubiquitin inhibitor. The Src kinase, Lck, as well as mitogen-activated protein kinase and Akt were activated, but p21WAF, phosphatase and tensin homologue was reduced in K562 BMS-R cells. Removal of dasatinib from the culture medium of K562 BMS-R cells led to apoptosis, and activated caspase 3 and poly (ADP-ribose) polymerase.

Conclusion: These results suggest that the expression and protein activation signatures identified in this study provide insight into the mechanism of resistance to dasatinib and imatinib and may be of therapeutic chronic myelogenous leukemia value clinically.
Brivanib Alaninate, a Dual Inhibitor of Vascular Endothelial Growth Factor Receptor and Fibroblast Growth Factor Receptor Tyrosine Kinases, Induces Growth Inhibition in Mouse Models of Human Hepatocellular Carcinoma
Authors: Huynh Hung , Ngo Van Chanh , Fargnoli Joseph , Ayers Mark , Soo Khee Chee , Koong Heng Nung , Thng Choon Hua , Ong Hock Soo , Chung Alexander , Chow Pierce , Pollock Pamela , Byron Sara , Tran Evelyn
Published: October 2008

Purpose: Hepatocellular carcinoma (HCC) is the fifth most common primary neoplasm; surgery is the only curative option but 5-year survival rates are only 25% to 50%. Vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF) are known to be involved in growth and neovascularization of HCC. Therefore, agents that target these pathways may be effective in the treatment of HCC. The aim of this study was to determine the antineoplastic activity of brivanib alaninate, a dual inhibitor of VEGF receptor (VEGFR) and FGF receptor (FGFR) signaling pathways.

Experimental Design: Six different s.c. patient-derived HCC xenografts were implanted into mice. Tumor growth was evaluated in mice treated with brivanib compared with control. The effects of brivanib on apoptosis and cell proliferation were evaluated by immunohistochemistry. The SK-HEP1 and HepG2 cells were used to investigate the effects of brivanib on the VEGFR-2 and FGFR-1 signaling pathways in vitro. Western blotting was used to determine changes in proteins in these xenografts and cell lines.

Results: Brivanib significantly suppressed tumor growth in five of six xenograft lines. Furthermore, brivanib–induced growth inhibition was associated with a decrease in phosphorylated VEGFR-2 at Tyr1054/1059, increased apoptosis, reduced microvessel density, inhibition of cell proliferation, and down-regulation of cell cycle regulators. The levels of FGFR-1 and FGFR-2 expression in these xenograft lines were positively correlated with its sensitivity to brivanib-induced growth inhibition. In VEGF-stimulated and basic FGF stimulated SK-HEP1 cells, brivanib significantly inhibited VEGFR-2, FGFR-1, extracellular signal-regulated kinase 1/2, and Akt phosphorylation.

Conclusion: This study provides a strong rationale for clinical investigation of brivanib in patients with HCC.
N-(2,6-Dimethoxypyridine-3-yl)-9-Methylcarbazole-3-Sulfonamide as a Novel Tubulin Ligand against Human Cancer
Authors: Wang Yue-Ming , Hu Lai-Xing , Liu Zhen-Ming , You Xue-Fu , Zhang Sheng-Hua , Qu Jing-Rong , Li Zhuo-Rong , Li Yan , Kong Wei-Jia , He Hong-Wei , Shao Rong-Guang , Zhang Liang-Ren , Peng Zong-Gen , Boykin David W. , Jiang Jian-Dong
Published: October 2008

Purpose: We have synthesized a new tubulin ligand N-(2,6-dimethoxypyridine-3-yl)-9-methylcarbazole-3-sulfonamide (IG-105). This work investigates its anticancer effect and mechanism.

Experimental Design: Anticancer efficacy was evaluated at the molecular target, cancer cells and nude mice. The mechanism was explored at submolecular, molecular, and cellular levels.

Results: IG-105 showed a potent activity against human leukemia and solid tumors in breast, liver, prostate, lung, skin, colon, and pancreas with IC50 values between 0.012 and 0.298 μmol/L. It was also active in drug-resistant tumor cells and not a P-glycoprotein substrate. It inhibited microtubule assembly followed by M-phase arrest, Bcl-2 inactivation, and then apoptosis through caspase pathways. The colchicine pocket on tubulin is the binding site of IG-105. Nude mice experiments showed that IG-105 monotherapy at 100 mg/kg i.p. (q2d) yielded 81% inhibition of Bel-7402 hepatoma growth and at 275 mg/kg i.p. (q2d) completely inhibited the tumor growth. MCF-7 breast cancer in nude mice showed a similar therapeutic response to IG-105. Acute toxicity of IG-105 was not found even at 1,000 mg/kg i.p. In combination with oxaliplatin or doxorubicin, IG-105 converted each of these subcurative compounds into a curative treatment with complete inhibition for tumor growth in the hepatoma-bearing nude mice. The combination was more active than either drug. In no experiment was toxicity increased by combination chemotherapy.

Conclusions: IG-105 inhibits microtubule assembly by binding at colchicine pocket. It shows a potent anticancer activity in vitro and in vivo and has good safety in mice. We consider IG-105 merits further investigation.
Circulating Tumor Cells Predict Survival Benefit from Treatment in Metastatic Castration-Resistant Prostate Cancer
Authors: de Bono Johann S. , Scher Howard I. , Montgomery R. Bruce , Parker Christopher , Miller M. Craig , Tissing Henk , Doyle Gerald V. , Terstappen Leon W.W.M. , Pienta Kenneth J. , Raghavan Derek
Published: October 2008

Purpose: A method for enumerating circulating tumor cells (CTC) has received regulatory clearance. The primary objective of this prospective study was to establish the relationship between posttreatment CTC count and overall survival (OS) in castration-resistant prostate cancer (CRPC). Secondary objectives included determining the prognostic utility of CTC measurement before initiating therapy, and the relationship of CTC to prostate-specific antigen (PSA) changes and OS at these and other time points.

Experimental Design: Blood was drawn from CRPC patients with progressive disease starting a new line of chemotherapy before treatment and monthly thereafter. Patients were stratified into predetermined Favorable or Unfavorable groups (<5 and ≥5 CTC/7.5mL).

Results: Two hundred thirty-one of 276 enrolled patients (84%) were evaluable. Patients with Unfavorable pretreatment CTC (57%) had shorter OS (median OS, 11.5 versus 21.7 months; Cox hazard ratio, 3.3; P < 0.0001). Unfavorable posttreatment CTC counts also predicted shorter OS at 2 to 5, 6 to 8, 9 to 12, and 13 to 20 weeks (median OS, 6.7-9.5 versus 19.6-20.7 months; Cox hazard ratio, 3.6-6.5; P < 0.0001). CTC counts predicted OS better than PSA decrement algorithms at all time points; area under the receiver operator curve for CTC was 81% to 87% and 58% to 68% for 30% PSA reduction (P = 0.0218). Prognosis for patients with (a) Unfavorable baseline CTC who converted to Favorable CTC improved (6.8 to 21.3 months); (b) Favorable baseline CTC who converted to Unfavorable worsened (>26 to 9.3 months).

Conclusions: CTC are the most accurate and independent predictor of OS in CRPC. These data led to Food and Drug Administration clearance of this assay for the evaluation of CRPC.
Dissociation of Its Opposing Immunologic Effects Is Critical for the Optimization of Antitumor CD8+ T-Cell Responses Induced by Interleukin 21
Authors: Ansén Sascha , Butler Marcus O. , Berezovskaya Alla , Murray Andrew P. , Stevenson Kristen , Nadler Lee M. , Hirano Naoto
Published: October 2008

Purpose: Interleukin 21 (IL-21) is a promising new cytokine, which is undergoing clinical testing as an anticancer agent. Although IL-21 provides potent stimulation of CD8+ T cells, it has also been suggested that IL-21 is immunosuppressive by counteracting the maturation of dendritic cells. The dissociation of these two opposing effects may enhance the utility of IL-21 as an immunotherapeutic. In this study, we used a cell-based artificial antigen-presenting cell (aAPC) lacking a functional IL-21 receptor (IL-21R) to investigate the immunostimulatory properties of IL-21.

Experimental Design: The immunosuppressive activity of IL-21 was studied using human IL-21R+ dendritic cells. Antigen-specific CD8+ T cells stimulated with human cell–based IL-21R-aAPC were used to isolate the T-cell immunostimulatory effects of IL-21. The functional outcomes, including phenotype, cytokine production, proliferation, and cytotoxicity were evaluated.

Results: IL-21 limits the immune response by maintaining immunologically immature dendritic cells. However, stimulation of CD8+ T cells with IL-21R- aAPC, which secrete IL-21, results in significant expansion. Although priming in the presence of IL-21 temporarily modulated the T-cell phenotype, chronic stimulation abrogated these differences. Importantly, exposure to IL-21 during restimulation promoted the enrichment and expansion of antigen-specific CD8+ T cells that maintained IL-2 secretion and gained enhanced IFN-γ secretion. Tumor antigen-specific CTL generated in the presence of IL-21 recognized tumor cells efficiently, demonstrating potent effector functions.

Conclusions: IL-21 induces opposing effects on antigen-presenting cells and CD8+ T cells. Strategic application of IL-21 is required to induce optimal clinical effects and may enable the generation of large numbers of highly avid tumor-specific CTL for adoptive immunotherapy.
Colon Cancer: It's CIN or CIMP
Authors: Issa Jean-Pierre
Published: October 2008

Combined genetic and epigenetic analysis of sporadic colon cancer suggest that it can no longer be viewed as a single disease. There are at least three different subsets with distinct clinico-pathologic features, with important implications for preventions, screening, and therapy.
Blood Loss during Transvaginal Oocyte Retrieval
Authors: Ragni Guido , Scarduelli Claudia , Calanna Graziella , Santi Giulia , Benaglia Laura , Somigliana Edgardo
Published: October 2008

Background/Aims: Limited information is available regarding the quantity of blood loss associated with uncomplicated transvaginal oocyte retrieval. The aim of the present study was evaluating the quantity of such a loss. Methods: One hundred and fifty consecutive women undergoing oocyte retrieval were recruited. They underwent blood test assessment and ultrasonographic transvaginal evaluation at three different times: (1) immediately before initiating oocyte retrieval, (2) 4–6 h later, and (3) 72 h later. Results: At 4–6 h after oocyte retrieval, the red blood cell count and the hemoglobin concentration were significantly reduced, whereas pelvic free fluid had significantly increased. The estimated median (Interquartile range) blood loss was 72 (–8/162) ml. None of the recruited women was found to have a hemoglobin reduction >2 g/day or an increase in the pelvic free fluid >200 ml or a calculated blood loss >500 ml (0.0%, 95% CI: 0.0–2.4%). No significant worsening from baseline was observed at the 72 h evaluation. Conclusions: The quantity of blood loss following oocyte retrieval is clinically unremarkable in the vast majority of women.
Back Pain Symptoms and Bone Mineral Density Changes in Pregnancy as Measured by Quantitative Ultrasound
Authors: To William W.K. , Wong Margaret W.N.
Published: October 2008

Objective: To evaluate whether back pain symptoms in pregnancy are associated with bone mineral density (BMD) changes as measured by quantitative ultrasound at the os calcis. Methods: Consecutive patients were recruited from a general obstetric clinic over 1 year. Quantitative ultrasonographic measurements of BMD were performed at the os calcis between weeks 14–20 and 36–38. They were then surveyed for back pain symptoms experienced during pregnancy in the early postpartum period, and these were subsequently correlated with BMD changes. Results: Of a total of 463 patients, 231 (49.8%) reported one or more episodes of significant back pain during pregnancy. A mean fall in BMD of 0.0395 g/cm2 was demonstrable from early to late gestation. Those with back pain symptoms have higher mean BMD loss at the os calcis during pregnancy compared to those without back pain (0.038 vs. 0.029 g/cm2; p = 0.012). A logistic regression model showed that weight gain and BMD loss in pregnancy were significantly associated with back pain symptoms. Conclusion: Back pain symptoms in pregnancy were associated with a greater fall in BMD at the os calcis, supporting the hypothesis that BMD changes could have an etiological role in back pain during pregnancy.
Completeness of Histopathology Reporting of Melanoma in a High-Incidence Geographical Region
Authors: Thompson B. , Austin R. , Coory M. , Aitken J.F. , Walpole E. , Francis G. , Fritschi L.
Published: October 2008

Background: Appropriate histopathology reporting helps to ensure effective therapy and prognosis. Objective: Toexamine compliance with clinical practice guidelines for histopathology reports of melanomas. Methods: A sample of melanoma histopathology reports in Queensland was audited for inclusion of recommended information. The quality of documentation was constructed and multivariate analysis used to determine factors affecting the quality of reporting practices. Results: Documentation of the most important features of melanoma was high: clear diagnosis (99.8%; 95% CI 98.6–100), thickness (99.8%; 95% CI 98.6–100), comment on adequacy of excision (87.9%; 95% CI 84.9–91.0) and measurement of margins (91.9%; 95% CI 88.8–91.4). Overall reporting of ulceration and regression was of lesser completeness (83.0 and 77.8%, respectively) and these features were more likely to be reported by high-volume laboratories (p < 0.001 and p = 0.037, respectively). This trend was not apparent for other features. Fewer than 50% of reports documented mitotic rate per square millimetre, predominant cell type, microsatellites, growth phase and desmoplasia. Conclusion: Awareness of current reporting practices and identification of areas in which insufficiencies exist enable the revision of systems and potential improvements to the transfer of information to treating clinicians.
IntroductionRethinking the Forms of Autonomy at the Dawn of the 21st Century
Authors: Légaré André , Suksi Markku
Published: September 2008

Arguably, other forms of autonomy also exist than territorial autonomy with law-making powers, such as personal autonomy, cultural autonomy and functional autonomy as well as autonomies with administrative or regulatory powers. The existence of such diff erent forms of autonomy should be kept in mind when the term ‘autonomy’ is used and the context of the usage of the term should be made clear. In addition, the interface between territorial autonomy with law-making powers on the one hand, and federal organization on the other is explored, the point being that a more coherent theoretical approach to territorial autonomy on the one hand and federalism on the other should be developed.
Stability and Change in Belief Systems
Authors: Renshon Jonathan
Published: October 2008

Cognitive frameworks and belief systems are the primary lenses through which presidents view the world. Two important questions are whether these beliefs ever change significantly, and if so, what causes these changes? This article develops empirical data on the strategic and operational beliefs of President George W. Bush (GWB) as a means of examining the theoretical basis for how and why core beliefs change. The author analyzes the foreign policy operational code of President Bush in four separate phases of his political career: the immediate prepresidential phase; his nine months in office prior to September 11, 2001; the six months immediately following the 9/11 terrorist attacks; and his last year in office as a second-term president. The results of this article allow us to address several recurring questions that concern the Bush administration and presidents' belief systems and decision making. They challenge traditional interpretations of the Bush presidency and provide some insights into the causal mechanisms that underlie belief change.
Pharmacokinetics and Pharmacodynamics of Dabigatran Etexilate, an Oral Direct Thrombin Inhibitor, Are Not Affected by Moderate Hepatic Impairment
Authors: Stangier Joachim , Stähle Hildegard , Rathgen Karin , Roth Willy , Shakeri‐Nejad Kasra
Published: October 2008

The impact of moderate hepatic impairment on the pharmacokinetics (PK) and pharmacodynamics (PD) of dabigatran etexilate was evaluated in an open, parallel‐group study. Healthy volunteers (n = 12) and patients with hepatic impairment (Child‐Pugh classification B; n = 12) received a single oral dose of 150 mg dabigatran etexilate. The mean values for area under the concentration‐time curve, terminal half‐life, and renal clearance of dabigatran were comparable between patients with hepatic impairment and healthy volunteers. Conversion of the dabigatran intermediate BIBR1087 to active dabigatran was slower in patients with hepatic impairment, indicating that the liver is partly involved in bioconversion of dabigatran etexilate. However, total drug exposure was comparable between groups; therefore, this observation is of no clinical relevance with respect to the anticoagulant activity of dabigatran. The extent of dabigatran glucuronidation was unchanged by liver disease; glucuronidation capacity was maintained in moderate liver disease. The activated partial thromboplastin time, ecarin clotting time, and thrombin time relationships were essentially identical in both groups. This study shows that moderate hepatic impairment does not affect the PK/PD or safety profile of dabigatran. Therefore, patients with moderate hepatic impairment can be given dabigatran etexilate without the need for dose adjustment.
Meta-analysis of Quality of Life Outcomes Following Diabetes Self-management Training
Authors: Cochran Jane , Conn Vicki S.
Published: October 2008

Purpose The purpose of this meta-analysis was to meet the need to quantify the influence of diabetes self-management training on quality of life (QOL) of adult diabetes patients.

Methods Extensive literature searching located published and unpublished diabetes self-management intervention studies that measured QOL outcomes among at least 5 subjects with type 1 or 2 diabetes. Data were extracted from primary study reports which included interventions designed to improve diabetes self-management and adequate data to calculate effect sizes. Random-effects meta-analytic procedures were used to estimate overall effects between treatment and control groups at outcome assessment and between baseline and outcome data for both treatment subjects and control subjects.

Results Exhaustive searching yielded 20 comparisons across 1892 subjects. The comparisons between treatment and control group outcomes following interventions yielded an effect size of 0.281. The comparisons between treatment group at baseline and outcome measurement yielded an effect size of 0.312 to 0.313. Each of these effect sizes were statistically significant, meaning that the hypothesis that interventions to improve diabetes self-management results in increased QOL was supported. Control subjects did not experience improved QOL while participating in studies.

Conclusions These findings document that people with diabetes experience improved QOL from participation in diabetes self-management training programs. Future diabetes self-management intervention studies should include quality of life outcomes so that this important outcome can be further studied. After more primary studies are available, future meta-analyses can explore important moderator analyses.
Dementia and Cognitive Impairment Are Not Associated With Earlier Mortality After Percutaneous Endoscopic Gastrostomy
Authors: Gaines David Isaac , Durkalski Valerie , Patel Alpesh , DeLegge Mark H.
Published: October 2008

Background: Studies have shown varying results with regard to risk factors for mortality after percutaneous endoscopic gastrostomy (PEG). Objectives: To examine the time to death in patients with dementia or significant cognitive impairment (SCI) due to neurologic injury who had undergone PEG compared with patients without either of these diagnoses, and to examine risk factors for 30‐day mortality after PEG. Methods: Patients who had undergone PEG over a 2‐year period were identified. Local medical records and the Social Security Death Index were reviewed to ascertain the patients' age, gender, serum albumin, diagnoses, presence or absence of dementia or SCI, presence or absence of complications related to PEG, and length of survival after PEG. The Charlson Comorbidity Index (CCI) was calculated based on the medical diagnoses at the time of PEG. Results: One hundred ninety patients were identified. Forty‐five carried a diagnosis of dementia and/or SCI compared with 145 who did not. Median survival of patients with dementia or SCI was 53 days compared with 78 days in patients without these diagnoses (P = .85). Age (odds ratio [OR] 1.1, 95% confidence interval [CI] 1.04‐1.12) and albumin (OR 0.43, 95% CI 0.22‐0.84) were associated with 30‐day mortality, whereas gender (OR 1.2, 95% CI 0.47‐2.90), CCI (OR 1.1, 95% CI 0.86‐1.32), and presence of PEG‐related complications (OR 1.6, 95% CI 0.36‐6.76) were not. Conclusions: Age and serum albumin are risk factors for 30‐day mortality after PEG. Patients with dementia or SCI do not have a significantly shorter survival after PEG than patients with intact cognitive function.
Variability in Weight and Histological Appearance of the Prostate of Beagle Dogs Used in Toxicology Studies
Authors: Dorso Laëtitia , Chanut Franck , Howroyd Paul , Burnett Roger
Published: October 2008

This review was performed to assess variations in weight and histologic appearance of the prostate of untreated male beagle dogs between 23 and 108 weeks of age, from two breeding centers. Data from 125 control beagle dogs from twenty-seven regulatory toxicology studies were used. Age, terminal body weight, and prostate weight were analyzed. Prostate sections were examined microscopically, and histological changes—such as development of acini, amount of secretion, and patterns of dilation and inflammation—were recorded and graded when appropriate. The influence of age, terminal body weight, and source on the weight and histological appearance of the prostate, and the degree of interanimal variation were evaluated
N-Methyl-N-Nitrosourea (MNU): A Positive Control Chemical for p53+/− Mouse Carcinogenicity Studies
Authors: Morton Daniel , Bailey Keith L. , Stout Cheryl L. , Weaver R. John , White Kathleen A. , Lorenzen Marianne J. , Ball Douglas J.
Published: October 2008

This study evaluated the effects of a single intraperitoneal injection of N-methyl- N-nitrosourea (MNU) in citrate buffer (pH 4.5) at a dose of 75 mg/kg in thirty male and thirty female p53+/− mice followed by a six-month observation period. Fifteen control mice per sex received a single intraperitoneal injection of citrate buffer. Fifty-six of sixty mice treated with MNU died or were sacrificed before the end of the observation period. Twenty-four males and twenty-seven females treated with MNU developed malignant lymphoma of the thymus; of these, twenty-three males and twenty-seven females had corresponding enlargement or masses in the thymus at necropsy. Lymphoblasts in thymic lymphomas stained positively for mouse CD3 antigen, indicating a T-cell lineage. One control female mouse had malignant lymphoma of the spleen that did not involve the thymus. Nine males and five females treated with MNU had adenomas or adenocarcinomas of the small intestine, whereas no intestinal neoplasms were observed in control mice. These findings support the use of a single dose of MNU as a positive control chemical in six-month p53+/− mouse carcinogenicity studies and suggest that examination of the thymus alone is sufficient to evaluate the validity of the model system.