bluuebunny/crossref_metadata_2025 huggingface dataset:
Segmentation of Striatal Brain Structures from High Resolution PET Images
Authors: Farinha Ricardo J. P. C. , Ruotsalainen Ulla , Hirvonen Jussi , Tuominen Lauri , Hietala Jarmo , Fonseca José M. , Tohka Jussi
Published: November 2009

We propose and evaluate an automatic segmentation method for extracting striatal brain structures (caudate, putamen, and ventral striatum) from parametric 11C‐raclopride positron emission tomography (PET) brain images. We focus on the images acquired using a novel brain dedicated high‐resolution (HRRT) PET scanner. The segmentation method first extracts the striatum using a deformable surface model and then divides the striatum into its substructures based on a graph partitioning algorithm. The weighted kernelk‐means algorithm is used to partition the graph describing the voxel affinities within the striatum into the desired number of clusters. The method was experimentally validated with synthetic and real image data. The experiments showed that our method was able to automatically extract caudate, ventral striatum, and putamen from the images. Moreover, the putamen could be subdivided into anterior and posterior parts. An automatic method for the extraction of striatal structures from high‐resolution PET images allows for inexpensive and reproducible extraction of the quantitative information from these images necessary in brain research and drug development.
A Case of Dilated Cardiomyopathy Associated with 3-Hydroxy-3-Methylglutaryl-Coenzyme A (HMG CoA) Lyase Deficiency
Authors: Leung Alexander A. C. , Chan Alicia K. , Ezekowitz Justin A. , Leung Alexander K. C.
Published: November 2009

3-hydroxy-3-methylglutaryl-coenzyme A (HMG CoA) lyase deficiency is an inborn error of metabolism characterized by impairment of ketogenesis and leucine catabolism resulting in an organic acidopathy. In 1994, a case of dilated cardiomyopathy and fatal arrhythmia was reported in a 7-month-old infant. We report a case of dilated cardiomyopathy in association with HMG CoA lyase deficiency in a 23-year-old man with the acute presentation of heart failure. To our knowledge, this is the first case reported in an adult.
The Potential for Xanthine Oxidase Inhibition in the Prevention and Treatment of Cardiovascular and Cerebrovascular Disease
Authors: Higgins Peter , Dawson Jesse , Walters Matthew
Published: November 2009

There is a now a wealth of epidemiological, animal, and clinical data to suggest the benefits of uric acid reduction and xanthine oxidase inhibition in prevention of vascular disease. This review discusses the available epidemiological, preclinical, and clinical data and considers arguments for and against a role for serum uric acid in common cardiovascular disorders. It concludes that large scale trials with clinical endpoints are justified to address this important question and to define whether use of drugs such as allopurinol should be a routine part of preventative strategies.
Genetic Polymorphisms in Genes Related to Oxidative Stress (GSTP1, GSTM1, GSTT1, CAT, MnSOD, MPO, eNOS) and Survival of Rectal Cancer Patients after Radiotherapy
Authors: Funke Silvia , Risch Angela , Nieters Alexandra , Hoffmeister Michael , Stegmaier Christa , Seiler Christoph M. , Brenner Hermann , Chang-Claude Jenny
Published: November 2009

Radiotherapy exerts part of its antineoplastic effect by generating oxidative stress, therefore genetic variation in oxidative stress-related enzymes may influence survival of rectal cancer patients. We hypothesized that genetic polymorphisms associated with higher amounts of reactive oxygen species (ROS) that exaggerate cytotoxic activity could improve survival after radiotherapy. We followed 114 rectal cancer patients who received radiotherapy for an average of 42.5 months. Associations between genotypes (GSTP1,GSTM1,GSTT1,CAT,MnSOD,MPOandeNOS) and overall survival were assessed using Kaplan-Meier curves and Cox proportional hazards regression. As hypothesized, patients carrying low ROS producingeNOSGlu298Asp asparagine allele showed an increased hazard of death compared to homozygous carriers of the glutamine allele (hazard ratio (HR): 2.10, 95% confidence interval (CI): 1.01–4.38). However, carriers of low ROS producingMPOG463A A allele had a decreased hazard of death compared to patients homozygous for the G allele (HR: 0.44, 95% CI: 0.21–0.93) although patients homozygous for the A allele had a slightly increased hazard (HR: 1.12, 95% CI: 0.25–5.08). This explorative study provides first results and highlights the need for further, larger studies to investigate association between genetic variation in oxidative stress genes and survival of rectal cancer patients who received radiotherapy.
Influence of Gap Size and Position within Gaps on Light Levels
Authors: Gendreau-Berthiaume Benoit , Kneeshaw Daniel
Published: November 2009

The previous studies have reported maximum light levels at different positions within gaps but many of these studies are based on gaps of different size. The objective of this study is to evaluate the influence of gap size and position within gap on light distribution in gaps and under the canopy north of gaps in a mixedwood temperate forest. For three gap sizes, with gap widths ranging between 0.5 and 1.5 times the height of the surrounding stand, light was measured at different positions along the north-south axis in each gap using two different techniques (hemispherical photographs and instantaneous measurements). In small gaps, the position with the most light was close to the northern edge although not under the canopy north of the gap. For both methods, the position with the highest light level shifted from the north towards the center of gaps as gap size increased which clarifies some of the variability in light measurements previously observed in gap studies. Higher light levels in the northern part of small and medium gaps compared to the southern portion could allow management of a mixture of species with intolerant species in the northern portions of gaps and tolerant species to the south.
Methyl‐CpG‐Binding PCR of Bloodspots for Confirmation of Fragile X Syndrome in Males
Authors: Tzeng Ching-Cherng , Liou Chiou-Ping , Li Chien-Feng , Lai Ming-Chi , Tsai Li-Ping , Cho Wei-Chen , Chang Hui-Ting
Published: November 2009

This study demonstrates that methyl‐CpG‐binding PCR (MB‐PCR) is a rapid and
simple method for detecting fragile X syndrome (FXS) in males, which is
performed by verifying the methylation status of the FMR1 promoter in
bloodspots. Proteins containing methyl‐CpG‐binding (MB) domains can be
freeze‐stored and used as stocks, and the entire test requires only a few hours.
The minimum amount of DNA required for the test is 0.5 ng. At this amount,
detection sensitivity is not hampered, even mixing with excess unmethylated
alleles up to 320 folds. We examined bloodspots from 100 males, including 24
with FXS, in a blinded manner. The results revealed that the ability of MB‐PCR to
detect FMR1 promoter methylation was the same as that of Southern blot
hybridization. Since individuals with 2 or more X chromosomes generally have
methylated FMR1 alleles, MB‐PCR cannot be used to detect FXS in females.
Advancements in Development of Chemical-Looping Combustion: A Review
Authors: Fang He , Haibin Li , Zengli Zhao
Published: November 2009

Chemical-looping combustion (CLC) is a novel combustion technology with inherent separation of greenhouseCO2. Extensive research has been performed on CLC in the last decade with respect to oxygen carrier development, reaction kinetics, reactor design, system efficiencies, and prototype testing. Transition metal oxides, such as Ni, Fe, Cu, and Mn oxides, were reported as reactive species in the oxygen carrier particles. Ni-based oxygen carriers exhibited the best reactivity and stability during multiredox cycles. The performance of the oxygen carriers can be improved by changing preparation method or by making mixedoxides. The CLC has been demonstrated successfully in continuously operated prototype reactors based on interconnected fluidized-bed system in the size range of 0.3–50 kW. High fuel conversion rates and almost 100%  CO2capture efficiencies were obtained. The CLC system with two interconnected fluidized-bed reactors was considered the most suitable reactor design. Development of oxygen carriers with excellent reactivity and stability is still one of the challenges for CLC in the near future. Experiences of building and operating the large-scale CLC systems are needed before this technology is used commercially. Chemical-looping reforming (CLR) and chemical-looping hydrogen (CLH) are novel chemical-looping techniques to produce synthesis gas and hydrogen deserving more attention and research.
An Inverse Eigenvalue Problem for Damped Gyroscopic Second‐Order Systems
Authors: Yuan Yongxin
Published: November 2009

The inverse eigenvalue problem of constructing symmetric positive semidefinite matrix
D (written as D ≥ 0) and real‐valued skew‐symmetric matrix G (i.e., GT = −G) of order n for the quadratic pencil Q(λ): = λ2Ma + λ(D + G) + Ka, where Ma > 0, Ka ≥ 0 are given
analytical mass and stiffness matrices, so that Q(λ) has a prescribed subset of eigenvalues and eigenvectors, is considered. Necessary and sufficient conditions under which this quadratic inverse eigenvalue problem is solvable are specified.
Isometries of a Bergman‐Privalov‐Type Space on the Unit Ball
Authors: Stević Stevo , Ueki Sei-Ichiro
Published: November 2009

We introduce a new space ANlog ,α(𝔹) consisting of all holomorphic functions on the unit ball 𝔹 ⊂ ℂn such that , where α > −1, (dV(z) is the normalized Lebesgue volume measure on 𝔹, and cα,n is a normalization constant, that is, Vα(𝔹) = 1), and φe(t) = tln (e + t) for t ∈ [0, ∞). Some basic properties of this space are presented. Among other results we proved that ANlog ,α(𝔹) with the metric is an F‐algebra with respect to pointwise addition and multiplication. We also prove that every linear isometry T of ANlog ,α(𝔹) into itself has the form Tf = c(f∘ψ) for some c ∈ ℂ such that |c| = 1 and some ψ which is a holomorphic self‐map of 𝔹 satisfying a measure‐preserving property with respect to the measure dVα. As a consequence of this result we obtain a complete characterization of all linear bijective isometries of ANlog ,α(𝔹).
Late Thrombosis of Sirolimus-Eluting Stent: A Multifactorial Problem
Authors: Kranjec Igor , Cerne Andreja
Published: November 2009

We report a case of a young patient in whom a sirolimus-eluting stent was implanted on the culprit left anterior descending coronary artery at primary percutaneous coronary intervention (PCI) for acute myocardial infarction. Nine months later she suffered from a reinfarction due to the late stent thrombosis despite a continuous antiplatelet therapy with aspirin and clopidogrel. A cluster of factors that might have contributed to the development of the stent thrombosis were identified: suboptimal PCI technique, complete stent fracture, and clopidogrel resistance. The obstructed stent was successfully reopened by repeat PCI, while the clopidogrel maintenance dosage was doubled to 150 mg daily for the following year. The further long-term clinical course was uneventful.
Urocortin in Second Trimester Amniotic Fluid: Its Role as Predictor of Preterm Labor
Authors: Iavazzo C. , Tassis K. , Gourgiotis D. , Boutsikou M. , Baka S. , Hassiakos D. , Vogiatzi C. , Florentin-Arar L. , Malamitsi-Puchner A.
Published: November 2009

Backgound. The existence of a “placental clock” which determines the duration of gestation has been previously proposed. It is related to placental CRH secretion and is active from an early phase in human pregnancy. Urocortin is a specific ligand for the corticotropin-releasing factor (CRF) receptor expressed by human trophoblast and fetal membranes. The purpose of this study was to evaluate whether urocortin concentrations in the early second trimester amniotic fluid might serve to predict preterm delivery.Method. The urocortin concentrations in early second trimester amniotic fluid were measured in 41 pregnancies with term delivery and in 41 pregnancies with preterm delivery by using an immunoradiometric assay. Conditional logistic regression analysis was used for statistical analysis.Results. Mean amniotic fluid urocortin concentrations in women with preterm labor were1.55±0.63 ng/mL while those in women with term labor were1.6±0.49 ng/mL (p: NS). No statistical significant results were found when comparing amniotic fluid urocortin concentrations in women with preterm premature rupture of membranes leading to preterm labor (n=19) to women with term delivery without premature rupture of membranes.Conclusion. These results suggest that urocortin concentrations in the amniotic fluid of genetic amniocentesis are not predictive of preterm labor and birth.
Specific Differential Equations for Generating Pulse Sequences
Authors: Toma Ghiocel
Published: November 2009

This study presents nonlinear differential equations capable to generate continuous functions similar to pulse sequences. First are studied some basic properties of second‐order differential equations with time‐dependent coefficients generating bounded oscillating functions F similar to test‐functions (the function F and its derivative F′ being equal to zero at the same time moments). The necessary intercorrelations between the phase of generated oscillations and the time‐dependent coefficients is presented, being shown also that the external command function should be set to a constant value at these time moments so as to determine the amplitude and the sign of generated oscillations. Then some possibilities of using previous differential equation for generating positive‐definite functions with null values for the function and its derivative at the same time moments and with constant slope for its amplitude are presented, being shown that the corresponding external command function should present also alternating components. Finally all previous results are used for determining a set of second‐order differential equations with time dependent coefficients and a set of external command and corrective functions for generating a pulse sequence useful for modelling time series.
Lin−Sca-1+CD49fhigh Stem/Progenitors Are Tumor-Initiating Cells in the Pten-Null Prostate Cancer Model
Authors: Mulholland David J. , Xin Li , Morim Ashkan , Lawson Devon , Witte Owen , Wu Hong
Published: November 2009

We have shown previously that Pten deletion leads to the expansion of subset of prostate cancer cells positive for CK5 and p63. Although this subpopulation may be involved in tumor initiation or progression, studies to date have not functionally validated this hypothesis. Using in vitro sphere-forming assay and in vivo prostate reconstitution assay, we show here the presence of a tumor-initiating subpopulation in the Pten prostate cancer mouse model. Specifically, we show that the Lin−Sca-1+CD49fhigh (LSC) subpopulation overlaps with CK5+;p63+ cells and is significantly increased during prostate cancer initiation and progression and after castration. Mutant spheres mimic the structural organization of the epithelial compartment in the Pten-null primary tumor. Sorted LSC cells from either Pten-null spheres or primary tumors are able to regenerate prostate epithelial structure with cancerous morphology, closely mimicking that of primary cancers. Therefore, the LSC subpopulation is capable of initiating a cancerous phenotype that recapitulates the pathology seen in the primary lesions of the Pten mutant prostate model. [Cancer Res 2009;69(22):8555–62]
Microenvironmental Independence Associated with Tumor Progression
Authors: Anderson Alexander R.A. , Hassanein Mohamed , Branch Kevin M. , Lu Jenny , Lobdell Nichole A. , Maier Julie , Basanta David , Weidow Brandy , Narasanna Archana , Arteaga Carlos L. , Reynolds Albert B. , Quaranta Vito , Estrada Lourdes , Weaver Alissa M.
Published: November 2009

Tumor-microenvironment interactions are increasingly recognized to influence tumor progression. To understand the competitive dynamics of tumor cells in diverse microenvironments, we experimentally parameterized a hybrid discrete-continuum mathematical model with phenotypic trait data from a set of related mammary cell lines with normal, transformed, or tumorigenic properties. Surprisingly, in a resource-rich microenvironment, with few limitations on proliferation or migration, transformed (but not tumorigenic) cells were most successful and outcompeted other cell types in heterogeneous tumor simulations. Conversely, constrained microenvironments with limitations on space and/or growth factors gave a selective advantage to phenotypes derived from tumorigenic cell lines. Analysis of the relative performance of each phenotype in constrained versus unconstrained microenvironments revealed that, although all cell types grew more slowly in resource-constrained microenvironments, the most aggressive cells were least affected by microenvironmental constraints. A game theory model testing the relationship between microenvironment resource availability and competitive cellular dynamics supports the concept that microenvironmental independence is an advantageous cellular trait in resource-limited microenvironments. [Cancer Res 2009;69(22):8797–806]
Retinoblastoma Family Proteins Have Distinct Functions in Pulmonary Epithelial Cells In vivo Critical for Suppressing Cell Growth and Tumorigenesis
Authors: Simpson David S. , Mason-Richie Nicole A. , Gettler Caitlin A. , Wikenheiser-Brokamp Kathryn A.
Published: November 2009

Lung cancer is the leading cause of cancer deaths, accounting for more deaths than breast, colon, and prostate cancer combined. The retinoblastoma (Rb)/p16 tumor suppressive pathway is deregulated in most cancers. Loss of p16 occurs more frequently than Rb loss, suggesting that p16 suppresses cancer by regulating Rb as well as the related proteins p107 and p130. However, direct evidence demonstrating that p130 or p107 cooperate with Rb to suppress epithelial cancers associated with p16 loss is currently lacking. Moreover, the roles of p130 and p107 in lung cancer are not clear. In the present studies, Rb ablation was targeted to the lung epithelium in wild-type, p107, or p130 null mice to determine unique and overlapping Rb family functions critical in tumor suppression. Rb ablation during development resulted in marked epithelial abnormalities despite p107 upregulation. In contrast, p130 and p107 were not required during development but had distinct functions in the Rb-deficient epithelium: p107 was required to suppress proliferation, whereas a novel proapoptotic function was identified for p130. Adult Rb-ablated lungs lacked the epithelial phenotype seen at birth and showed compensatory p107 upregulation and p16 induction in epithelial cell lineages that share phenotypic characteristics with human non–small cell lung cancers (NSCLC) that frequently show p16 loss. Importantly, Rb/p107-deficient, but not Rb/p130-deficient, lungs developed tumors resembling NSCLC. Taken together, these studies identify distinct Rb family functions critical in controlling epithelial cell growth, and provide direct evidence that p107 cooperates with Rb to protect against a common adult cancer. [Cancer Res 2009;69(22):8733–41]
Inhibition of Rho-Associated Kinase Signaling Prevents Breast Cancer Metastasis to Human Bone
Authors: Liu Sijin , Goldstein Robert H. , Scepansky Ellen M. , Rosenblatt Michael
Published: November 2009

Rho-associated kinase (ROCK) signaling plays a fundamental role in regulating cell morphology, adhesion, and motility. Aberrant expression of ROCK is related to tumor metastases and poor clinical outcome. Here, we show that ROCK expression is increased in metastatic human mammary tumors and breast cancer cell lines compared with nonmetastatic tumors and cell lines. Overexpression of ROCK confers a metastatic phenotype on the nonmetastatic MCF-7 cell line. Inhibition of ROCK activity, by either a specific ROCK inhibitor (Y27632) or ROCK-targeted small interfering RNAs, reduces cell migration and proliferation in vitro and metastasis to bone in vivo using a novel “human breast cancer metastasis to human bone” mouse model. Expression of the c-Myc–regulated miR-17-92 cluster is shown to be elevated in metastatic breast cancer cells compared with nonmetastatic cells and diminished by Y27632 treatment. Furthermore, blockade of miR-17 is shown to decrease breast cancer cell invasion/migration in vitro and metastasis in vivo. Together, these findings suggest that augmented ROCK signaling contributes to breast cancer metastasis. The effects of ROCK on tumor cell invasion/motility and growth may derive from regulating cytoskeletal actin-myosin contraction and modulating the c-Myc pathway, including c-Myc–dependent microRNAs. Inhibition of ROCK or the pathway it stimulates, therefore, may represent a novel approach for treatment of breast cancer metastases. [Cancer Res 2009;69(22):8742–51]
Aurora B–Mediated Phosphorylation of RASSF1A Maintains Proper Cytokinesis by Recruiting Syntaxin16 to the Midzone and Midbody
Authors: Song Su Jung , Kim Soon Jung , Song Min Sup , Lim Dae-Sik
Published: November 2009

Aurora B is critically involved in ensuring proper cytokinesis and maintaining genomic stability. The tumor suppressor RASSF1A regulates cell cycle progression by regulating mitotic progression, G1-S transition, and microtubule stability. We previously reported that both Aurora A and Aurora B phosphorylate RASSF1A, and showed that phosphorylation of RASSF1A by Aurora A blocks the inhibitory function of RASSF1A toward anaphase-promoting complex-Cdc20. However, the role of Aurora B–mediated RASSF1A phosphorylation remains unknown. Here, we show that phosphorylation of RASSF1A on Ser203 by Aurora B during late mitosis has a critical role in regulating cytokinesis. Notably, RASSF1A interacts with Syntaxin16, a member of the t-SNARE family, at the midzone and midbody during late mitosis. Aurora B is required for this interaction and for the subsequent recruitment of Syntaxin16 to the midzone and midbody, a prerequisite for the successful completion of cytokinesis. Furthermore, Aurora B depletion results in a failure of Syntaxin16 to properly localize to the midzone and midbody, a mislocalization that was prevented by overexpression of the phosphomimetic RASSF1A (S203D) mutant. Finally, either depletion of Syntaxin16 or expression of the nonphosphorylatable RASSF1A (S203A) mutant results in cytokinesis defects. Our findings implicate Aurora B–mediated phosphorylation of RASSF1A in the regulation of cytokinesis. [Cancer Res 2009;69(22):8540–4]
Pattern of Antioxidant and DNA Repair Gene Expression in Normal Airway Epithelium Associated with Lung Cancer Diagnosis
Authors: Blomquist Thomas , Crawford Erin L. , Mullins D'Anna , Yoon Youngsook , Hernandez Dawn-Alita , Khuder Sadik , Ruppel Patricia L. , Peters Elizabeth , Oldfield David J. , Austermiller Brad , Anders John C. , Willey James C.
Published: November 2009

In previous studies, we reported that key antioxidant and DNA repair genes are regulated differently in normal bronchial epithelial cells of lung cancer cases compared with non–lung cancer controls. In an effort to develop a biomarker for lung cancer risk, we evaluated the transcript expressions of 14 antioxidant, DNA repair, and transcription factor genes in normal bronchial epithelial cells (HUGO names CAT, CEBPG, E2F1, ERCC4, ERCC5, GPX1, GPX3, GSTM3, GSTP1, GSTT1, GSTZ1, MGST1, SOD1, and XRCC1). A test comprising these 14 genes accurately identified the lung cancer cases in two case-control studies. The receiver operating characteristic–area under the curve was 0.82 (95% confidence intervals, 0.68–0.91) for the first case-control set (25 lung cancer cases and 24 controls), and 0.87 (95% confidence intervals, 0.73–0.96) for the second set (18 cases and 22 controls). For each gene included in the test, the key difference between cases and controls was altered distribution of transcript expression among cancer cases compared with controls, with more lung cancer cases expressing at both extremes among all genes (Kolmorogov-Smirnov test, D = 0.0795; P = 0.041). A novel statistical approach was used to identify the lower and upper boundaries of transcript expression that optimally classifies cases and controls for each gene. Based on the data presented here, there is an increased prevalence of lung cancer diagnosis among individuals that express a threshold number of key antioxidant, DNA repair, and transcription factor genes at either very high or very low levels in the normal airway epithelium. [Cancer Res 2009;69(22):8629–35]
TP53 Cancerous Mutations Exhibit Selection for Translation Efficiency
Authors: Waldman Yedael Y. , Tuller Tamir , Sharan Roded , Ruppin Eytan
Published: November 2009

The tumor suppressor gene TP53 is known to be a key regulator in cancer, and more than half of human cancers exhibit mutations in this gene. Recent evidence shows that point mutations in TP53 not only disrupt its function but also possess gain-of-function and dominant-negative effects on wild-type copies, thus making the mutated gene an oncogene. Hence, this brings about the possibility that TP53 mutations may be under selection for increasing the overall translation efficiency (TE) of defected TP53 in cancerous cells. Here, we perform the first large-scale analysis of TE in human cancer mutated TP53 variants, identifying a significant increase in TE that is correlated with the frequency of TP53 mutations. Furthermore, mutations with a known oncogenic effect significantly increase their TE compared with the other TP53 mutations. Further analysis shows that TE may have influence both on selecting the location of the mutation and on its outcome: codons with lower TE show stronger selection toward nonsynonymous mutations and, for each codon, frequent mutations show stronger increase in TE compared with less frequent mutations. Additionally, we find that TP53 mutations have significantly higher TE increase in progressive versus primary tumors. Finally, an analysis of TP53 NCI-60 cell lines points to a coadaptation between the mutations and the tRNA pool, increasing the overall TP53 TE. Taken together, these results show that TE plays an important role in the selection of TP53 cancerous mutations. [Cancer Res 2009;69(22):8807–13]
A Novel Role of Interleukin-13 Receptor α2 in Pancreatic Cancer Invasion and Metastasis
Authors: Fujisawa Toshio , Joshi Bharat , Nakajima Atsushi , Puri Raj K.
Published: November 2009

Whereas interleukin-13 receptor α2 chain (IL-13Rα2) is overexpressed in a variety of human solid cancers including pancreatic cancer, we investigated its significance in cancer invasion and metastasis. We used two pancreatic cancer cell lines, IL-13Rα2–negative HPAF-II and IL-13Rα2–positive HS766T, and generated IL-13Rα2 stably transfected HPAF-II as well as IL-13Rα2 RNA interference knocked-down HS766T cells. Ability of invasion and signal transduction was compared between IL-13Rα2–negative and IL-13Rα2–positive cells and tumor metastasis was assessed in murine model for human pancreatic cancer with orthotopic implantation of tumors. IL-13 treatment enhanced cell invasion in IL-13Rα2–positive cancer cell lines but not in IL-13Rα2–negative cell lines. Furthermore, gene transfer of IL-13Rα2 in negative cell lines enhanced invasion, whereas its silencing downmodulated invasion of pancreatic cell lines in a Matrigel invasion assay. In vivo study revealed that IL-13Rα2–positive cancer metastasized to lymph nodes, liver, and peritoneum at a significantly higher rate compared with IL-13Rα2–negative tumors. The expression of IL-13Rα2 in metastatic lesions was found to be increased compared with primary tumors, and mice with IL-13Rα2–positive cancer displayed cachexia and poor prognosis. Invasion and metastasis also correlated with increased matrix metalloproteinase protease activity in these cells. Mechanistically, IL-13 activated extracellular signal-regulated kinase 1/2 and activator protein-1 nuclear factors in IL-13Rα2–positive pancreatic cancer cell lines but not in IL-13Rα2–negative cell lines. Taken together, our results show for the first time that IL-13 can signal through IL-13Rα2 in pancreatic cancer cells and IL-13Rα2 may serve as a prognostic biomarker of invasion and metastasis in pancreatic cancer. [Cancer Res 2009;69(22):8678–85]
Phase I/II Trial of Bevacizumab and Radiotherapy for Locally Advanced Inoperable Colorectal Cancer: Vasculature-Independent Radiosensitizing Effect of Bevacizumab
Authors: Koukourakis Michael I. , Giatromanolaki Alexandra , Sheldon Helen , Buffa Francesca M. , Kouklakis George , Ragoussis Ioannis , Sivridis Efthimios , Harris Adrian L. ,   for the Tumour and Angiogenesis Research Group
Published: November 2009

Purpose: Antivascular endothelial growth factor therapy enhances the activity of radiotherapy in experimental models, and bevacizumab has therapeutic activity in patients with metastatic colorectal cancer.

Experimental Design: Twenty-two patients with locally advanced inoperable colorectal carcinomas (LA/I-CRC) were treated with conformal hypofractionated (3.4 Gy/fraction 15) split-course accelerated radiotherapy (biological equivalent dose, 67.2 Gy) supported with amifostine, capecitabine (600 mg/m2 daily, 5 days/week), and bevacizumab (5 mg/kg every 2 weeks, five cycles). Biopsies from nine patients, performed before and 1 week after bevacizumab administration, were analyzed for changes in mRNA expression with Illumina gene arrays.

Results: No serious grade 3 chemotherapy-related side effects were recorded. There was low acute toxicity, with moist perineal desquamation noted in 2 of 22 patients, diarrhea grade 2 to 3 in 5 of 22 patients, and severe proctalgia in 2 of 22 patients. One patient died from Fournier's gangrene before treatment completion. Within a median follow-up of 18 months, two patients with preradiotheraphy direct involvement of adjacent organs expressed recto-vaginal/perineal fistula. Out of 19 evaluable cases, 13 (68.5) showed complete response and 4 showed (21.1) partial response. Fourteen patients are alive with no evidence of loco-regional relapse. In the gene array analysis, 30 known genes associated with transcription factors, DNA repair, and proliferation were downregulated by bevacizumab. DUSP1 gene was the most consistently downregulated transcript.

Conclusions: The combination of radiotherapy with bevacizumab is feasible and results in a high rate of durable complete responses in patients with LA/I-CRC. Radiosensitization may occur through a direct effect on tumor cells followed by a wide scale suppression of transcription factors and genes involved in DNA repair and proliferation. (Clin Cancer Res 2009;15(22):706976)
Production of Interferon-Inducible Protein-10 and Its Role as an Autocrine Invasion Factor in Nasal Natural Killer/T-Cell Lymphoma Cells
Authors: Moriai Shigetaka , Takahara Miki , Ogino Takeshi , Nagato Toshihiro , Kishibe Kan , Ishii Hideyuki , Katayama Akihiro , Shimizu Norio , Harabuchi Yasuaki
Published: November 2009

Purpose: Nasal natural killer (NK)/T-cell lymphoma is associated with Epstein-Barr virus and has poor prognosis because of local invasion and/or multiple dissemination. Recently, the role of chemokines/chemokine receptors in tumor proliferation and invasion has been shown. In this study, we examined whether the specific chemokines were related to the tumor behaviors in nasal NK/T-cell lymphoma.

Experimental Design: A chemokine protein array was used to examine specific chemokines produced by SNK-6 and SNT-8 (Epstein-Barr viruspositive nasal NK/T-cell lymphoma lines). The expression of interferon inducible protein 10 (IP-10) and the IP-10 receptor CXCR3 was investigated by ELISA and flow cytometry. Cell growth and invasion were assessed by the MTT and Matrigel invasion assays, respectively. Immunohistologic staining and ELISA were used to examine IP-10 expression in biopsies and sera from patients, respectively.

Results: IP-10 was specifically produced by SNK-6 and SNT-8. Moreover, CXCR3 was expressed on the NK cell lines. Functionally, IP-10 did not affect cell proliferation but enhanced cell invasion. In biopsy samples, IP-10 and CXCR3 expressions were detected in the lymphoma cells. Serum IP-10 levels in the patients were much higher than those of healthy controls and the levels were decreased during the complete remission phase after treatments.

Conclusions: These results suggest that IP-10 may play an important role in cell invasion in nasal NK/T-cell lymphoma through an autocrine mechanism. (Clin Cancer Res 2009;15(22):67719)
Circulating Tumor Cell as a Diagnostic Marker in Primary Lung Cancer
Authors: Tanaka Fumihiro , Yoneda Kazue , Kondo Nobuyuki , Hashimoto Masaki , Takuwa Teruhisa , Matsumoto Seiji , Okumura Yoshitomo , Rahman Shakibur , Tsubota Noriaki , Tsujimura Tohru , Kuribayashi Kozo , Fukuoka Kazuya , Nakano Takashi , Hasegawa Seiki
Published: November 2009

Purpose: To investigate the diagnostic performance of circulating tumor cells (CTC) in discrimination between primary lung cancer and nonmalignant diseases as well as in prediction of distant metastasis.

Patients and Methods: We prospectively evaluated CTCs in 7.5-mL samples of peripheral blood sampled from patients with a suspicion or a diagnosis of primary lung cancer. A semiautomated system was used to capture CTCs with an antibody against epithelial cell adhesion molecule.

Results: Of 150 eligible patients, 25 were finally diagnosed as having nonmalignant disease, and 125 were diagnosed as having primary lung cancer with (n = 31) or without (n = 94) distant metastasis. CTCs were detected in 30.6 of lung cancer patients and in 12.0 of nonmalignant patients. CTC count was significantly higher in lung cancer patients than in nonmalignant patients, but a receiver operating characteristic (ROC) curve analysis showed an insufficient capability of the CTC test in discrimination between lung cancer and nonmalignant diseases with an area under ROC curve of 0.598 (95 confidence interval, 0.488-0.708; P = 0.122). Among lung cancer patients, CTC count significantly increased along with tumor progression, especially with development of distant metastasis. The area under ROC curve for CTC count in prediction of distant metastasis was 0.783 (95 confidence interval, 0.679-0.886; P < 0.001). When patients with one or more CTCs were judged as having metastatic disease, sensitivity and specificity of the CTC test were 71.0 and 83.0, respectively.

Conclusions: CTC is a useful surrogate marker of distant metastasis in primary lung cancer. (Clin Cancer Res 2009;15(22):69806)
A Single Supratherapeutic Dose of Vorinostat Does Not Prolong the QTc Interval in Patients with Advanced Cancer
Authors: Munster Pamela N. , Rubin Eric H. , Van Belle Simon , Friedman Evan , Patterson Jaclyn K. , Van Dyck Kristien , Li Xiadong , Comisar Wendy , Chodakewitz Jeffery A. , Wagner John A. , Iwamoto Marian
Published: November 2009

Purpose: This dedicated QTc phase I study, conducted in advanced-stage cancer patients, assessed the effect of a single supratherapeutic dose (800 mg) of vorinostat on the QTc interval.

Experimental Design: A randomized, partially blind, placebo-controlled, two-period, crossover study was conducted. Patients (n = 25) received single doses of 800 mg vorinostat and placebo in the fasted state. Holter electrocardiogram monitoring was done before each treatment and for 24 h postdose. Blood samples for vorinostat concentration were collected through 24 h postdose following vorinostat treatment only. Prescribed electrocardiogram and blood sampling times were designed to capture the expected Cmax of vorinostat.

Results: Twenty-four of the 25 patients enrolled in the study were included in the QTc analysis. The upper bound of the two-sided 90 confidence interval for the QTcF interval for the placebo-adjusted mean change from baseline of vorinostat was <10 ms at every time point. No patient had a QTcF change from baseline value >30 ms. One patient had QTcF values >450 ms (seen after both vorinostat and placebo administration) and none had values >480 ms. Mean AUC0- and Cmax values attained were on the order of 1.93- and 1.41-fold higher, respectively, compared with the 400 mg clinical dose. Based on assessment of clinical and laboratory adverse experiences, single doses of 800 mg vorinostat were generally well tolerated.

Conclusions: Administration of a single supratherapeutic dose of the histone deacetylase inhibitor vorinostat is not associated with prolongation of the QTc interval. A dedicated QTc study in advanced cancer patients is a robust means for assessing risk for ventricular repolarization prolongation. (Clin Cancer Res 2009;15(22):707784)
Randomized Biomarker Trial of Anastrozole or Low-Dose Tamoxifen or Their Combination in Subjects with Breast Intraepithelial Neoplasia
Authors: Bonanni Bernardo , Serrano Davide , Gandini Sara , Guerrieri-Gonzaga Aliana , Johansson Harriet , Macis Debora , Cazzaniga Massimiliano , Luini Alberto , Cassano Enrico , Oldani Sabina , Lien Ernst A. , Pelosi Giuseppe , Decensi Andrea
Published: November 2009

Purpose: In the Anastrozole, Tamoxifen Alone or in Combination trial, the combination arm was inferior to anastrozole alone in terms of disease-free survival possibly due to an adverse pharmacokinetic interaction or a predominant estrogenic effect of tamoxifen under estrogen deprivation. We assessed whether the addition of a lower dose of tamoxifen influenced anastrozole bioavailability and favorably modulated biomarkers of bone fracture, breast cancer, cardiovascular disease, and endometrial cancer risk. The influence of CYP2D6 genotype on tamoxifen effects was also determined.

Experimental Design: Seventy-five postmenopausal women with breast intraepithelial neoplasia were randomly allocated to either 1 mg/d anastrozole or 10 mg/wk tamoxifen or their combination for 12 months. Study endpoints were plasma drug concentrations and changes of C-telopeptide, osteocalcin, estradiol/sex hormone binding globulin (SHBG) ratio, estrone sulfate, insulin-like growth factor-I (IGF-I)/insulin-like growth factor binding protein-3 (IGFBP-3), C-reactive protein, antithrombin-III, endometrial Ki-67 expression, and thickness.

Results: Anastrozole concentrations were not affected by the combination with low-dose tamoxifen, whereas endoxifen levels were lower in poor CYP2D6 metabolizers. C-telopeptide increased by 20 with anastrozole and decreased by 16 with tamoxifen and by 7 with their combination (P < 0.001); osteocalcin showed similar changes. Compared with anastrozole, the combination arm showed lower IGF-I/IGFBP-3 levels (17 versus 9; P = 0.004) and lower estradiol/SHBG and estrone sulfate reductions (15 versus 29 and 30 versus 38, respectively). However, IGF-I/IGFBP-3 and estradiol/SHBG did not decrease in poor CYP2D6 metabolizers. Endometrial thickness was not greater in the combination than in the anastrozole arm.

Conclusions: The addition of a weekly tamoxifen administration did not impair anastrozole bioavailability and modulated favorably its safety profile, providing the rationale for further studies. (Clin Cancer Res 2009;15(22):705360)
Role of Class 3 Semaphorins and Their Receptors in Tumor Growth and Angiogenesis
Authors: Gaur Puja , Bielenberg Diane R. , Samuel Shaija , Bose Debashish , Zhou Yunfei , Gray Michael J. , Dallas Nikolaos A. , Fan Fan , Xia Ling , Lu Jia , Ellis Lee M.
Published: November 2009

Class 3 semaphorins (SEMA3) were first identified as glycoproteins that negatively mediate neuronal guidance by binding to neuropilin and repelling neurons away from the source of SEMA3. However, studies have shown that SEMA3s are also secreted by other cell types, including tumor cells, where they play an inhibitory role in tumor growth and angiogenesis (specifically SEMA3B and SEMA3F). SEMA3s primarily inhibit the cell motility and migration of tumor and endothelial cells by inducing collapse of the actin cytoskeleton via neuropilins and plexins. Besides binding to SEMA3s, neuropilin also binds the protumorigenic and proangiogenic ligand vascular endothelial growth factor (VEGF). Although some studies attribute the antitumorigenic and antiangiogenic properties of SEMA3s to competition between SEMA3s and VEGF for binding to neuropilin receptors, several others have shown that SEMA3s display growth-inhibitory activity independent of competition with VEGF. A better understanding of these molecular interactions and the role and signaling of SEMA3s in tumor biology will help determine whether SEMA3s represent potential therapeutic agents. Herein, we briefly review (a) the role of SEMA3s in mediating tumor growth, (b) the SEMA3 receptors neuropilins and plexins, and (c) the potential competition between SEMA3s and VEGF family members for neuropilin binding. (Clin Cancer Res 2009;15(22):676370)
Synergistic antitumor effect between vorinostat and topotecan in small cell lung cancer cells is mediated by generation of reactive oxygen species and DNA damage-induced apoptosis
Authors: Bruzzese Francesca , Rocco Monia , Castelli Silvia , Di Gennaro Elena , Desideri Alessandro , Budillon Alfredo
Published: November 2009

The topoisomerase-I (topo-I) inhibitor topotecan, derivative of camptothecin, is the only registered drug for relapsed small cell lung cancer (SCLC). The histone deacetylase inhibitor vorinostat has shown preclinical and clinical antitumor activities in hematologic malignancies and solid tumors, including SCLC, and has recently been approved for the treatment of cutaneous T-cell lymphomas. In this study, we analyzed the antitumor effect of vorinostat combined with topotecan or camptothecin in topo-I inhibitor-sensitive H209 and inhibitor-resistant H526 SCLC cells. Simultaneous or sequential exposure (24 h delay) to either agent resulted in strong synergistic cytotoxic effect in both cell lines, as shown by calculating combination index, and confirmed by growth in soft agar. Combination treatments increased S-phase cell cycle arrest paralleled by apoptosis as measured by hypodiploid peak formation, Annexin V binding, DNA fragmentation, and mitochondria destruction. The apoptotic process was triggered by a caspase-dependent mechanism and can be ascribed to the phosphorylation of H2AX, a reporter of DNA double-strand breaks. These effects were paralleled by an increase of topo-I/DNA covalent complexes induced by combination treatment and suggest a potentiation by vorinostat of topotecan-induced DNA damage. Finally, oxidative injury played a significant functional role in the observed enhanced lethality because coadministration of the antioxidant N-acetyl-l-cysteine blocked reactive oxygen species generation, apoptosis, and mitochondria destruction induced by the vorinostat/topotecan combination. To our knowledge, this is the first demonstration of a synergistic antitumor effect between topotecan and vorinostat in SCLC. Because no well-established treatment is available for recurrent SCLC patients, our results indicate that this drug combination should be explored clinically. [Mol Cancer Ther 2009;8(11):3075–87]
Experimental cancer therapy using restoration of NAD+-linked 15-hydroxyprostaglandin dehydrogenase expression
Authors: Kaliberova Lyudmila N. , Kusmartsev Sergei A. , Krendelchtchikova Valentina , Stockard Cecil R. , Grizzle William E. , Buchsbaum Donald J. , Kaliberov Sergey A.
Published: November 2009

Preclinical and clinical evidence shows that cyclooxygenase-2 (Cox-2)-mediated prostaglandin E2 (PGE2) overexpression plays an important role in tumor growth, metastasis, and immunosuppression. It has been shown that expression of NAD+-linked 15-hydroxyprostaglandin dehydrogenase (15-PGDH), a key enzyme responsible for PGE2 inactivation, is suppressed in the majority of cancers, including breast and colon carcinoma. We have developed adenoviral vectors (Ad) encoding the 15-PGDH gene under control of the vascular endothelial growth factor receptor 1 (VEGFR1/flt-1; Adflt-PGDH) and the Cox-2 (Adcox-PGDH) promoters. The purpose of this study was to investigate cytotoxicity in vitro and therapeutic efficacy in vivo of 15-PGDH–mediated cancer therapy. The levels of PGE2 and VEGF expression were correlated with PGE2 receptor and Cox-2 and flt-1 expression in cancer cells. The in vitro study showed that Ad-mediated 15-PGDH expression significantly decreased proliferation and migration of cancer cells. Animal breast and colon tumor therapy studies showed that 15-PGDH gene therapy produced a significant delay in 2LMP and LS174T tumor growth. Combined therapy using 15-PGDH and anti-VEGF antibody (bevacizumab) significantly increased inhibition of growth of LS174T tumor xenografts in comparison with agents alone. These results suggest that 15-PGDH–mediated regulation of PGE2 catabolism in the tumor microenvironment represents a novel approach for therapy of human breast and colon cancer. [Mol Cancer Ther 2009;8(11):3130–9]
Positive selection of gene-modified cells increases the efficacy of pancreatic cancer suicide gene therapy
Authors: Martinez-Quintanilla Jordi , Cascallo Manel , Gros Alena , Fillat Cristina , Alemany Ramon
Published: November 2009

Thymidine kinase (TK)-mediated suicide gene therapy has been considered for the treatment of pancreatic cancer. However, despite a bystander effect, the proportion of transduced tumor cells has proven too low to result in efficacy. We propose the use of a drug-selectable marker (MDR1) to enrich TK-expressing cells using chemotherapy. This enrichment or positive selection phase may increase the efficacy of suicide gene therapy. To test this strategy, we generated stable NP18MDR/TK-GFP transfectants and showed docetaxel resistance in vivo. Mixed tumors of MDR/TK-expressing cells and parental NP18 cells were established and docetaxel was used to increase the proportion of TK-expressing cells. After this positive selection phase, suicide gene therapy with ganciclovir was applied. Upon positive selection, the proportion of TK-expressing cells increased from 4% to 22%. Subsequent suicide gene therapy was more effective compared with a control group without positive selection. Starting with 10% of TK-expressing cells the positive-negative selection strategy completely inhibited tumor growth. Taken together, these results suggest that a positive-negative selection strategy based on MDR and TK genes represents an efficient way to increase the proportion of TK-expressing cells in the tumor and the efficacy of TK-mediated suicide gene therapy. [Mol Cancer Ther 2009;8(11):3098–107]
Characterization of caspase-dependent and caspase-independent deaths in glioblastoma cells treated with inhibitors of the ubiquitin-proteasome system
Authors: Foti Carmela , Florean Cristina , Pezzutto Antonio , Roncaglia Paola , Tomasella Andrea , Gustincich Stefano , Brancolini Claudio
Published: November 2009

The regulation of the necrotic death and its relevance in anticancer therapy are largely unknown. Here, we have investigated the proapoptotic and pronecrotic activities of two ubiquitin-proteasome system inhibitors: bortezomib and G5. The present study points out that the glioblastoma cell lines U87MG and T98G are useful models to study the susceptibility to apoptosis and necrosis in response to ubiquitin-proteasome system inhibitors. U87MG cells show resistance to apoptosis induced by bortezomib and G5, but they are more susceptible to necrosis induced by G5. Conversely, T98G cells are more susceptible to apoptosis induced by both inhibitors but show some resistance to G5-induced necrosis. No overt differences in the induction of Noxa and Mcl-1 or in the expression levels of other components of the apoptotic machinery were observed between U87MG and T98G cells. Instead, by comparing the transcriptional profiles of the two cell lines, we have found that the resistance to G5-induced necrosis could arise from differences in glutathione synthesis/utilization and in the microenvironment. In particular, collagen IV, which is highly expressed in T98G cells, and fibronectin, whose adhesive function is counteracted by tenascin-C in U87MG cells, can restrain the necrotic response to G5. Collectively, our results provide an initial characterization of the molecular signals governing cell death by necrosis in glioblastoma cell lines. [Mol Cancer Ther 2009;8(11):3140–50]