bluuebunny/crossref_metadata_2025 huggingface dataset:
Construction of a Recombinant Eukaryotic Expression Plasmid Containing Human Calcitonin Gene and Its Expression in NIH3T3 Cells
Authors: Li Xiaolin , Jiang Guozhong , Wu Dan , Wang Xiuli , Zeng Bingfang
Published: August 2009

Aim. To construct a recombinant eukaryotic expression plasmid containing human calcitonin (hCT) gene and express the gene in murine fibroblast NIH3T3 cells. Materials and Methods. A murine Igκ‐chain leader sequence and hCT gene were synthesized and cloned into pCDNA3.0 to form the pCDNA3.0‐Igκ‐hCT eukaryotic expression vector, which was transfected into NIH3T3 cells. The mRNA and protein expressions and secretion of hCT were detected. Primarily cultured osteoclasts were incubated with the supernatant of pCDNA3.0‐Igk‐hCT‐transfected NIH3T3 cells, and their numbers were counted and morphology observed. Results. The expression and secretion of hCT were successfully detected in pCDNA3.0‐Igk‐hCT‐transfected NIH3T3 cells. The number of osteoclasts was decreased and the cells became crumpled when they were incubated with the supernatant of pCDNA3.0‐Igk‐hCT‐transfected NIH3T3 cells. Conclusion. A recombinant eukaryotic expression vector containing hCT gene was successfully constructed and expressed in NIH3T3 cells. The secreted recombinant hCT inhibited the growth and morphology of osteoclasts.
Modelling of HTR Confinement Behaviour during Accidents Involving Breach of the Helium Pressure Boundary
Authors: Fontanet Joan , Herranz Luis E. , Ramlakan Alastair , Naicker Lolan
Published: August 2009

Development of HTRs requires the performance of a thorough safety study, which includes accident analyses. Confinement building performance is a key element of the system since the behaviour of aerosol and attached fission products within the building is of an utmost relevance in terms of the potential source term to the environment. This paper explores the available simulation capabilities (ASTEC and CONTAIN codes) and illustrates the performance of a postulated HTR vented confinement under prototypical accident conditions by a scoping study based on two accident sequences characterized by Helium Pressure Boundary breaches, a small and a large break. The results obtained indicate that both codes predict very similar thermal‐hydraulic responses of the confinement both in magnitude and timing. As for the aerosol behaviour, both codes predict that most of the inventory coming into the confinement is eventually depleted on the walls and only about 1% of the aerosol dust is released to the environment. The crosscomparison of codes states that largest differences are in the intercompartmental flows and the in‐compartment gas composition.
The Role of Computational Fluid Dynamics in the Management of Unruptured Intracranial Aneurysms: A Clinicians′ View
Authors: Singh Pankaj K. , Marzo Alberto , Coley Stuart C. , Berti Guntram , Bijlenga Philippe , Lawford Patricia V. , Villa-Uriol Mari-Cruz , Rufenacht Daniel A. , McCormack Keith M. , Frangi Alejandro , Patel Umang J. , Hose D. Rodney
Published: August 2009

Objective. The importance of hemodynamics in the etiopathogenesis of intracranial aneurysms (IAs) is widely accepted. Computational fluid dynamics (CFD) is being used increasingly for hemodynamic predictions. However, alogn with the continuing development and validation of these tools, it is imperative to collect the opinion of the clinicians.Methods. A workshop on CFD was conducted during the European Society of Minimally Invasive Neurological Therapy (ESMINT) Teaching Course, Lisbon, Portugal. 36 delegates, mostly clinicians, performed supervised CFD analysis for an IA, using the @neuFuse software developed within the European project @neurIST. Feedback on the workshop was collected and analyzed. The performance was assessed on a scale of 1 to 4 and, compared with experts′ performance.Results. Current dilemmas in the management of unruptured IAs remained the most important motivating factor to attend the workshop and majority of participants showed interest in participating in a multicentric trial. The participants achieved an average score of 2.52 (range 0–4) which was 63% (range 0–100%) of an expert user.Conclusions. Although participants showed a manifest interest in CFD, there was a clear lack of awareness concerning the role of hemodynamics in the etiopathogenesis of IAs and the use of CFD in this context. More efforts therefore are required to enhance understanding of the clinicians in the subject.
Valproic Acid-Induced Hyperammonemia in the Elderly: A Review of the Literature
Authors: Mittal Vikrant , Muralee Sunanda , Tampi Rajesh R.
Published: August 2009

Valproic acid and its derivatives are commonly used to treat many psychiatric conditions in the elderly. Hyperammonemia is a less common but important side effect of these drugs. The elderly patient appears highly vulnerable to this side effect of this group of medications. In this paper, we systematically review the published literature for hyperammonemia induced by valproic acid and its derivatives. We describe the three reported cases and review possible treatment strategies for this condition.
A New XOR Structure Based on Resonant-Tunneling High Electron Mobility Transistor
Authors: Sharifi Mohammad Javad , Bahrepour Davoud
Published: August 2009

A new structure for an exclusive-OR (XOR) gate based on the resonant-tunneling high electron mobility transistor (RTHEMT) is introduced which comprises only an RTHEMT and two FETs. Calculations are done by utilizing a new subcircuit model for simulating the RTHEMT in the SPICE simulator. Details of the design, input, and output values and margins, delay of each transition, maximum operating frequency, static and dynamic power dissipations of the new structure are discussed and calculated and the performance is compared with other XOR gates which confirm that the presented structure has a high performance. Furthermore, to the best of authors' knowledge, it has the least component count in comparison to the existing structures.
Absorption Line Profile of the Transition of Atomic Oxygen and Its Application to Plasma Monitoring
Authors: Sasaki K. , Okumura Y. , Asaoka R.
Published: August 2009

The line profile of the transition of atomic oxygen was measured by diode laser absorption spectroscopy. As a result, it was found that the absorption line profile had a wing component in the wavelength range detuned from the line center and was not fitted with a Gaussian function. The wing component was considered to be originated from dissociative excitation of
molecular oxygen. We fitted the absorption line profile with the superposition of two Gaussian
functions corresponding to high and low translational temperatures. We propose that the ratio of
the high-temperature to low-temperature components is useful for monitoring the relative degree of
dissociation of molecular oxygen in oxygen-containing plasmas. The ratio of the high-temperature
to low-temperature components was compared with the survival ratio of molecular oxygen, which
was evaluated from the lifetime of in the afterglow of pulsed discharges.
Sprouty2 Association with B-Raf Is Regulated by Phosphorylation and Kinase Conformation
Authors: Brady Suzanne C. , Coleman Mathew L. , Munro June , Feller Stephan M. , Morrice Nicolas A. , Olson Michael F.
Published: August 2009

Sprouty2 is a feedback regulator that controls the Ras/Raf/MEK/extracellular signal-regulated kinase mitogen-activated protein kinase (MAPK) pathway at multiple levels, one way being through direct interaction with Raf kinases. Consistent with a role as a tumor suppressor, Sprouty2 expression is often down-regulated in human cancers. However, Sprouty2 is up-regulated in some cancers, suggesting the existence of posttranscriptional mechanisms that permit evasion of Sprouty2-mediated antitumorigenic properties. We report that MAPK activation induces Sprouty2 phosphorylation on six serine residues, which reduced Sprouty2 association with wild-type B-Raf. Mutation of these six serines to nonphosphorylatable alanines increased the ability of Sprouty2 to inhibit growth factor–induced MAPK activation. Oncogenic B-Raf mutants such as B-Raf V600E did not associate with Sprouty2, but this resistance to Sprouty2 binding was not due to phosphorylation. Instead, the active kinase conformation induced by oncogenic mutation prevents Sprouty2 binding. These results reveal a dual mechanism that affects the Sprouty2/B-Raf interaction: Sprouty phosphorylation and B-Raf conformation. [Cancer Res 2009;69(17):6773–81]
Heat Shock Protein 90 Inhibitor 17-Dimethylaminoethylamino-17-Demethoxygeldanamycin Enhances EphA2+ Tumor Cell Recognition by Specific CD8+ T Cells
Authors: Kawabe Mayumi , Mandic Maja , Taylor Jennifer L. , Vasquez Cecilia A. , Wesa Amy K. , Neckers Leonard M. , Storkus Walter J.
Published: August 2009

EphA2, a member of the receptor tyrosine kinase family, is commonly expressed by a broad range of cancer types, where its level of (over)expression correlates with poor clinical outcome. Because tumor cell expressed EphA2 is a nonmutated “self” protein, specific CD8+ T cells are subject to self-tolerance mechanisms and typically exhibit only moderate-to-low functional avidity, rendering them marginally competent to recognize EphA2+ tumor cells in vitro or in vivo. We have recently reported that the ability of specific CD8+ T cells to recognize EphA2+ tumor cells can be augmented after the cancer cells are pretreated with EphA2 agonists that promote proteasomal degradation and up-regulated expression of EphA2/class I complexes on the tumor cell membrane. In the current study, we show that treatment of EphA2+ tumor cells with the irreversible heat shock protein 90 inhibitor, 17-dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG), similarly enhances their recognition by EphA2-specific CD8+ T-cell lines and clones in vitro via a mechanism that is dependent on proteasome and transporter-associated protein function as well as the retrotranslocation of EphA2 into the tumor cytoplasm. When 17-DMAG and agonist anti-EphA2 monoclonal antibodies are coapplied, T-cell recognition of tumor cells is further increased over that observed for either agent alone. These studies suggest that EphA2 represents a novel heat shock protein 90 client protein and that the treatment of cancer patients with 17-DMAG–based “pulse” therapy may improve the antitumor efficacy of CD8+ T effector cells reactive against EphA2-derived epitopes. [Cancer Res 2009;69(17):6995–7003]
Blockade of Wnt Signaling Inhibits Angiogenesis and Tumor Growth in Hepatocellular Carcinoma
Authors: Hu Jie , Dong Aiwen , Fernandez-Ruiz Veronica , Shan Juanjuan , Kawa Milosz , Martínez-Ansó Eduardo , Prieto Jesus , Qian Cheng
Published: August 2009

Aberrant activation of Wnt signaling plays an important role in hepatocarcinogenesis. In addition to direct effects on tumor cells, Wnt signaling might be involved in the organization of tumor microenvironment. In this study, we have explored whether Wnt signaling blockade by exogenous expression of Wnt antagonists could inhibit tumor angiogenesis and control tumor growth. Human Wnt inhibitory factor 1 (WIF1) and secreted frizzled-related protein 1 (sFRP1) were each fused with Fc fragment of human IgG1 to construct fusion proteins WIF1-Fc and sFRP1-Fc. The recombinant adenoviral vectors carrying WIF1-Fc and sFRP1-Fc driven by cytomegalovirus promoter were constructed. Ad-WIF1-Fc or Ad-sFRP1-Fc induced the expression and correct conformation of WIF1-Fc and sFRP1-Fc fusion proteins. These molecules caused down-regulation of E2F1, cyclin D1, and c-myc and promoted cell apoptosis in hepatocellular carcinoma cells. Treatment of established hepatocellular carcinoma tumors with Ad-WIF1-Fc and/or Ad-sFRP1-Fc resulted in significant inhibition of tumor growth and prolonged animal survival. The antineoplastic effect was associated with increased apoptosis of tumor cells, reduced microvessel density, and decreased expression of vascular endothelial growth factor and stromal cell–derived factor-1. Tube formation and migration of human microvascular endothelial cells and mouse endothelial progenitor cells (EPC) were significantly inhibited by both WIF1-Fc and sFRP1-Fc. In addition, these molecules blocked EPC differentiation and caused EPC apoptosis. Our data indicate that Wnt antagonists WIF1-Fc and sFRP1-Fc inhibit Wnt signaling and exert potent antitumor activity by increasing the apoptosis rate in tumor cells and by impairing tumor vascularization. [Cancer Res 2009;69(17):6951–59]
Proteolytic Cleavage of Protein Tyrosine Phosphatase μ Regulates Glioblastoma Cell Migration
Authors: Burgoyne Adam M. , Phillips-Mason Polly J. , Burden-Gulley Susan M. , Robinson Shenandoah , Sloan Andrew E. , Miller Robert H. , Brady-Kalnay Susann M.
Published: August 2009

Glioblastoma multiforme (GBM), the most common malignant primary brain tumor, represents a significant disease burden. GBM tumor cells disperse extensively throughout the brain parenchyma, and the need for tumor-specific drug targets and pharmacologic agents to inhibit cell migration and dispersal is great. The receptor protein tyrosine phosphatase μ (PTPμ) is a homophilic cell adhesion molecule. The full-length form of PTPμ is down-regulated in human glioblastoma. In this article, overexpression of full-length PTPμ is shown to suppress migration and survival of glioblastoma cells. Additionally, proteolytic cleavage is shown to be the mechanism of PTPμ down-regulation in glioblastoma cells. Proteolysis of PTPμ generates a series of proteolytic fragments, including a soluble catalytic intracellular domain fragment that translocates to the nucleus. Only proteolyzed PTPμ fragments are detected in human glioblastomas. Short hairpin RNA–mediated down-regulation of PTPμ fragments decreases glioblastoma cell migration and survival. A peptide inhibitor of PTPμ function blocks fragment-induced glioblastoma cell migration, which may prove to be of therapeutic value in GBM treatment. These data suggest that loss of cell surface PTPμ by proteolysis generates catalytically active PTPμ fragments that contribute to migration and survival of glioblastoma cells. [Cancer Res 2009;69(17):6960–8]
Distinct Concentration-Dependent Effects of the Polo-like Kinase 1–Specific Inhibitor GSK461364A, Including Differential Effect on Apoptosis
Authors: Gilmartin Aidan G. , Bleam Maureen R. , Richter Mark C. , Erskine Symon G. , Kruger Ryan G. , Madden Lenore , Hassler Daniel F. , Smith Gary K. , Gontarek Richard R. , Courtney Mary P. , Sutton David , Diamond Melody A. , Jackson Jeffrey R. , Laquerre Sylvie G.
Published: August 2009

Polo-like kinase 1 (Plk1) is a conserved serine/threonine kinase that plays an essential role in regulating the many processes involved in mitotic entry and progression. In humans, Plk1 is expressed primarily during late G2 and M phases and, in conjunction with Cdk1/cyclin B1, acts as master regulatory kinases for the myriad protein substrates involved in mitosis. Plk1 overexpression is strongly associated with cancer and has been correlated with poor prognosis in a broad range of human tumor types. We have identified a potent, selective, reversible, ATP-competitive inhibitor of Plk1, GSK461364A, capable of inhibiting cell growth of most proliferating cancer cell lines tested. We observe distinct cell cycle effects of GSK461364A depending on the dose used. The predominant phenotype for cells treated with GSK461364A is prometaphase arrest with characteristic collapsed polar polo spindle. At high concentrations, GSK461364A delays mitotic entry in G2 followed by gradual progression into terminal mitosis; in some cell lines, this correlates with decreased apoptosis. Cell culture growth inhibition by GSK461364A can be cytostatic or cytotoxic but leads to tumor regression in xenograft tumor models under proper dose scheduling. Finally, we describe pharmacodynamic biomarkers of GSK461364A activity (pHH3 and Plk1) that are currently being evaluated in human cancer clinical trials. [Cancer Res 2009;69(17):6969–77]
Functional Significance of CytochromeP450 1B1 in Endometrial Carcinogenesis
Authors: Saini Sharanjot , Hirata Hiroshi , Majid Shahana , Dahiya Rajvir
Published: August 2009

Cytochrome P450 1B1 (CYP1B1) catalyzes estrogen hydroxylation and activation of potential carcinogens. Here we explored the role of CYP1B1 in endometrial carcinogenesis. Immunohistochemical staining of endometrial carcinomas showed that CYP1B1 is up-regulated in endometrial cancers. To understand the functional significance of CYP1B1 up-regulation in endometrial cancers with regard to tumorigenesis, we used small interfering RNA–mediated approach to knockdown CYP1B1 in endometrial carcinoma cell line followed by functional assays. Further, to understand the molecular basis of the role of CYP1B1 in endometrial carcinomas, we profiled the expression of key pathway-specific genes and identified several components of cell cycle, apoptosis, and cell adhesion pathways that are potentially regulated by CYP1B1. CYP1B1 depletion in endometrial carcinoma cells leads to decreased cellular proliferation and induces G0-G1 cell cycle arrest. Significantly, CYP1B1 knockdown leads to down-regulated expression of cyclin E1, S-phase kinase-associated protein 2 (SKP2), minichromosome maintenance complex component 4 (MCM4), and RAD51 and up-regulation of p27Kip1. Also, we identified cyclin E–binding protein (CEBP1) as a novel CYP1B1 target. Attenuation of CYP1B1 expression in endometrial carcinoma cells induces apoptosis and increases expression of IFN-β (IFNβ), granzyme A (GRZA), and tumor necrosis factor–related apoptosis-inducing ligand (TRAIL). Importantly, CYP1B1 depletion decreased the invasive potential of the endometrial cancer cells and expression of melanoma cell adhesion molecule (MCAM). In conclusion, our data suggest that CYP1B1 up-regulation plays a crucial role in endometrial carcinogenesis by targeting multiple pathways. We speculate that CYP1B1 inhibition in endometrial carcinomas could be a useful therapeutic approach as it regulates several potential anticancer targets like cyclin E1, Skp2, and TRAIL. [Cancer Res 2009;69(17):7038–45]
Imaging Mass Spectrometry of a Specific Fragment of Mitogen-Activated Protein Kinase/Extracellular Signal-Regulated Kinase Kinase Kinase 2 Discriminates Cancer from Uninvolved Prostate Tissue
Authors: Cazares Lisa H. , Troyer Dean , Mendrinos Savvas , Lance Raymond A. , Nyalwidhe Julius O. , Beydoun Hind A. , Clements Mary Ann , Drake Richard R. , Semmes O. John
Published: August 2009

Purpose: Histopathology is the standard approach for tissue diagnostics and centerpiece of pathology. Although the current system provides prognostic information, there is need for molecular markers that enhance diagnosis and better predict clinical prognosis. The ability to localize disease-specific molecular changes in biopsy tissue would help improve critical pathology decision making. Direct profiling of proteins from tissue using matrix-assisted laser desorption/ionization imaging mass spectrometry has the potential to supplement morphology with underlying molecular detail.

Experimental Design: A discovery set of 11 prostate cancer (PCa)–containing and 10 benign prostate tissue sections was evaluated for protein expression differences. A separate validation set of 54 tissue sections (23 PCa and 31 benign) was used to verify the results. Cryosectioning was done to yield tissue sections analyzed by a pathologist to determine tissue morphology and mirror sections for imaging mass spectrometry. Spectra were acquired and the intensity of signals was plotted as a function of the location within the tissue.

Results: An expression profile was found that discriminates between PCa and normal tissue. The overexpression of a single ion at m/z 4,355 was able to discriminate cancer from uninvolved tissue. Tandem mass spectrometry identified this marker as a fragment of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase kinase 2 (MEKK2). The ability of MEKK2 to discriminate tumor from normal cells was orthogonally confirmed.

Conclusions: This study highlights the potential of this approach to uncover molecular detail that can be correlated with pathology decision making. In addition, the identification of MEKK2 shows the ability to discover proteins of relevance to PCa biology. (Clin Cancer Res 2009;15(17):5541–51)
15d-PGJ2 Induces Apoptosis by Reactive Oxygen Species–mediated Inactivation of Akt in Leukemia and Colorectal Cancer Cells and Shows In vivo Antitumor Activity
Authors: Shin Sung-Won , Seo Chi-Yeon , Han Hoon , Han Jin-Yeong , Jeong Jin-Sook , Kwak Jong-Young , Park Joo-In
Published: August 2009

Purpose: Recent studies have shown that 15-deoxy-Δ12, 14-prostaglandin J2 (15d-PGJ2), a natural ligand for peroxisome proliferator–activated receptor-γ (PPARγ), inhibits cell proliferation and induces apoptosis. The specific molecular mechanisms underlying this effect remain to be elucidated. We examined whether 15d-PGJ2 has antitumor activity in vitro and in vivo, and investigated the underlying mechanism.

Experimental Design: We examined 15d-PGJ2–induced apoptosis in human leukemia cells in the context of mitochondrial injury, oxidative damage, and signaling pathway disturbances. In addition, we investigated the antitumor effect of 15d-PGJ2 in a mouse CT-26 s.c. tumor model and HL-60 leukemia xenograft model.

Results: 15d-PGJ2 induced apoptosis in leukemia and colorectal cancer cells in a dose-dependent manner and led to generation of reactive oxygen species (ROS) through mitochondria and NADPH oxidase activation, activation of JNK, and inactivation of Akt, a serine/threonine-specific protein kinase. Constitutive activation of Akt for an engineered myristoylated protein prevented 15d-PGJ2–mediated apoptosis but not ROS generation. Collectively, these findings suggest a hierarchical model of apoptosis induced by 15d-PGJ2 in human leukemia cells: oxidative injury represents a primary event resulting in Akt inactivation, which in turn leads to mitochondrial injury and apoptosis. Moreover, 15d-PGJ2 markedly reduced growth of mouse CT-26 s.c. tumors and HL-60 xenograft tumors and down-regulated p-Akt and Akt expression in vivo.

Conclusions: These results suggest that Akt inactivation through ROS production may contribute to 15d-PGJ2–induced apoptosis in leukemia and colorectal cancer cell lines and that 15d-PGJ2 may have therapeutic relevance in the treatment of human leukemia and colorectal cancer. (Clin Cancer Res 2009;15(17):5414–25)
Differences in Repair of DNA Cross-links between Lymphocytes and Epithelial Tumor Cells from Colon Cancer Patients Measured In vitro with the Comet Assay
Authors: Herrera Mercedes , Dominguez Gemma , Garcia Jose M. , Peña Cristina , Jimenez Carmen , Silva Javier , Garcia Vanesa , Gomez Irene , Diaz Raquel , Martin Paloma , Bonilla Felix
Published: August 2009

Purpose: The more common approach to comet assay studies with cancer patients involves indirect measurement of the effect of antineoplastic drug or radiation regimen by assessing DNA damage in surrogate cells, such as peripheral blood lymphocytes of cancer patients, to predict how tumor cells may be affected. The aim of the present study was to compare the capability of different cells isolated from a series of 23 colon cancer patients to repair the damage induced by a cancer drug.

Experimental Design: We adapted the in vitro comet repair assay for nucleotide excision repair to measure the ability of lymphocytes and normal and tumor epithelial colon cells to remove DNA cross-links induced by oxaliplatin. The excision repair rate was measured quantitatively by the tail parameters: tail DNA, tail length, extent tail moment, and olive tail moment.

Results: Kruskal-Wallis analysis revealed significant differences in recognition and excision activity between different cell types (P < 0.001) for all the comet parameters studied. Hence, colon cells showed higher recognition and excision activity than lymphocytes and tumor cells displayed the highest repair capability. We found no significant correlation between the repair activity of tumor colon cells and lymphocytes in any of the comet parameters considered.

Conclusions: Our data support the view that lymphocyte repair activity is not predictive of the repair ability of the tumor and that lymphocytes cannot act as surrogate cells. (Clin Cancer Res 2009;15(17):5466–72)
Herbal Extract ofWedelia chinensisAttenuates Androgen Receptor Activity and Orthotopic Growth of Prostate Cancer in Nude Mice
Authors: Tsai Chin-Hsien , Lin Feng-Min , Yang Yu-Chih , Lee Ming-Ting , Cha Tai-Lung , Wu Guan-James , Hsieh Shih-Chuan , Hsiao Pei-Wen
Published: August 2009

Purpose: Wedelia chinensis is a common ingredient of anti-inflammatory herbal medicines in Taiwan and southern China. Inflammation is involved in promoting tumor growth, invasion, and metastasis. This study aims to test the biological effects in vivo of W. chinensis extract on prostate cancer.

Experimental Design: The in vivo efficacy and mechanisms of action of oral administration of a standardized extract of W. chinensis were analyzed in animals bearing a subcutaneous or orthotopic prostate cancer xenograft.

Results: Exposure of prostate cancer cells to W. chinensis extract induced apoptosis selectively in androgen receptor (AR)–positive prostate cancer cells and shifted the proportion in each phase of cell cycle toward G2-M phase in AR-negative prostate cancer cells. Oral herbal extract (4 or 40 mg/kg/d for 24-28 days) attenuated the growth of prostate tumors in nude mice implanted at both subcutaneous (31% and 44%, respectively) and orthotopic (49% and 49%, respectively) sites. The tumor suppression effects were associated with increased apoptosis and lower proliferation in tumor cells as well as reduced tumor angiogenesis. The antitumor effect of W. chinensis extract was correlated with accumulation of the principle active compounds wedelolactone, luteolin, and apigenin in vivo.

Conclusion: Anticancer action of W. chinensis extract was due to three active compounds that inhibit the AR signaling pathway. Oral administration of W. chinensis extract impeded prostate cancer tumorigenesis. Future studies of W. chinensis for chemoprevention or complementary medicine against prostate cancer in humans are thus warranted. (Clin Cancer Res 2009;15(17):5435–44)
mTOR Is a Promising Therapeutic Target Both in Cisplatin-Sensitive and Cisplatin-Resistant Clear Cell Carcinoma of the Ovary
Authors: Mabuchi Seiji , Kawase Chiaki , Altomare Deborah A. , Morishige Kenichirou , Sawada Kenjiro , Hayashi Masami , Tsujimoto Masahiko , Yamoto Mareo , Klein-Szanto Andres J. , Schilder Russell J. , Ohmichi Masahide , Testa Joseph R. , Kimura Tadashi
Published: August 2009

Purpose: Mammalian target of rapamycin (mTOR) plays a central role in cell proliferation and is regarded as a promising target in cancer therapy, including for ovarian cancer. This study aimed to examine the role of mTOR as a therapeutic target in clear cell carcinoma of the ovary, which is regarded as an aggressive, chemoresistant histologic subtype.

Experimental Design: Using tissue microarrays of 98 primary ovarian cancers (52 clear cell carcinomas and 46 serous adenocarcinomas), the expression of phospho-mTOR was assessed by immunohistochemistry. Then, the growth-inhibitory effect of mTOR inhibition by RAD001 (everolimus) was examined using two pairs of cisplatin-sensitive parental (RMG1 and KOC7C) and cisplatin-resistant human clear cell carcinoma cell lines (RMG1-CR and KOC7C-CR) both in vitro and in vivo.

Results: Immunohistochemical analysis showed that mTOR was more frequently activated in clear cell carcinomas than in serous adenocarcinomas (86.6% versus 50%). Treatment with RAD001 markedly inhibited the growth of both RMG1 and KOC7C cells both in vitro and in vivo. Increased expression of phospho-mTOR was observed in cisplatin-resistant RMG1-CR and KOC7C-CR cells, compared with the respective parental cells. This increased expression of phospho-mTOR in cisplatin-resistant cells was associated with increased activation of AKT. RMG1-CR and KOC7C-CR cells showed greater sensitivity to RAD001 than did parental RMG1 and KOC7C cells, respectively, in vitro and in vivo.

Conclusion: mTOR is frequently activated in clear cell carcinoma and can be a promising therapeutic target in the management of clear cell carcinoma. Moreover, mTOR inhibition by RAD001 may be efficacious as a second-line treatment of recurrent disease in patients previously treated with cisplatin. (Clin Cancer Res 2009;15(17):5404–13)
Mapatumumab, a Fully Human Agonistic Monoclonal Antibody That Targets TRAIL-R1, in Combination with Gemcitabine and Cisplatin: a Phase I Study
Authors: Mom Constantijne H. , Verweij Jaap , Oldenhuis Corina N.A.M. , Gietema Jourik A. , Fox Norma Lynn , Miceli Renée , Eskens Ferry A.L.M. , Loos Walter J. , de Vries Elisabeth G.E. , Sleijfer Stefan
Published: August 2009

Purpose: To evaluate the safety, tolerability, pharmacokinetics, and antitumor activity of mapatumumab, a fully human monoclonal antibody targeting tumor necrosis factor–related apoptosis–inducing ligand receptor 1 (TRAIL-R1), in combination with gemcitabine and cisplatin.

Experimental Design: Patients with advanced solid tumors received gemcitabine 1,250 mg/m2 i.v. on days 1 and 8 and cisplatin 80 mg/m2 i.v. on day 1 of each 21-day cycle. Escalating mapatumumab doses were administered i.v. every 21 days. Toxicity was evaluated and pharmacokinetic analysis of plasma mapatumumab, gemcitabine, 2-difluoro-2-deoxyuridine, and unbound and total platinum was done. TRAIL-R1 tumor expression was determined immunohistochemically.

Results: Forty-nine patients received mapatumumab (1 mg/kg, n = 4; 3 mg/kg, n = 7; 10 mg/kg, n = 12; 20 mg/kg, n = 13; or 30 mg/kg, n = 13). A median of six cycles (range, 1-48) was administered. The adverse events most commonly observed reflect the toxicity profile of gemcitabine and cisplatin. Dose-limiting toxicities were seen in 3 of 12 patients at 10 mg/kg, consisting of grade 3 transaminitis, neutropenic fever, and grade 4 thrombocytopenia. At 20 mg/kg, 2 of 12 patients had dose-limiting toxicities, including grade 4 thrombocytopenia and grade 4 fatigue. The maximum tolerated dose was not reached. Pharmacokinetic interactions have not been observed. Twelve patients had a partial response, and 25 patients showed stable disease with a median duration of 6 months.

Conclusions: Mapatumumab in combination with gemcitabine and cisplatin is safe and well tolerated at doses up to 30 mg/kg. Further studies on this combination are warranted. (Clin Cancer Res 2009;15(17):5584–90)
Is the Response to Growth Hormone in Short Children Born Small for Gestational Age Dependent on Genetic or Maternal Factors?
Authors: Mehls O. , Lindberg A. , Bettendorf M. , Doerr H.-G. , Hauffa B.P. , Kaspers S. , Rohrer T. , Stahnke N. , Ranke M.B.
Published: August 2009

Background/Aims: We investigated whether genetic or maternal/environmental risk factors for being born small for gestational age (SGA), e.g. Silver-Russell syndrome, congenital heart defects, infections of mothers or smoking during pregnancy, explain the variation in the first-year growth response to GH therapy. Methods: Secondary analysis was made of growth response in 135 short prepubertal German children (66% males) enrolled in a SGA phase III trial. Initial mean patient age was 6.8 ± 2.6 years; mean patient height SDS –3.8 ± 1.2, and GH treatment dose was 0.066 mg/kg body weight per day. Results: Growth velocity increased by 4.5 ± 2.0 cm/year and height SDS by 1.0 ± 0.5 SDS. Although patient number was limited and variation was high, both growth response (cm/year) and change in height SDS did not appear to differ between subgroups which also did not differ in terms of Studentized residuals set up in the KIGS growth prediction model for SGA. Likewise, in a step-forward multivariate analysis, the variables Silver-Russell syndrome, congenital heart defects, infections of mothers and smoking were not identified as independent factors influencing growth velocity. Conclusion: The retrospectively analyzed genetic and maternal/environmental risk factors for SGA do not appear to explain the observed patient variance in response to GH. Larger prospective studies are needed, however, to substantiate these preliminary findings.
Cohort Studies: Prospective versus Retrospective
Authors: Euser Anne M. , Zoccali Carmine , Jager Kitty J. , Dekker Friedo W.
Published: August 2009

Cohort studies form a suitable study design to assess associations between multiple exposures on the one hand and multiple outcomes on the other hand. They are especially appropriate to study rare exposures or exposures for which randomization is not possible for practical or ethical reasons. Prospective and retrospective cohort studies have higher accuracy and higher efficiency as their respective main advantages. In addition to possible confounding by indication, cohort studies may suffer from selection bias. Confounding and bias should be prevented whenever possible, but still can exert unknown effects in unknown directions. If one is aware of this, cohort studies can form a potent study design in nephrology producing, in general, highly generalizable results.
Expression of Tumor Necrosis Factor Receptors on Granulocytes in Patients with Myeloperoxidase Anti-Neutrophil Cytoplasmic Autoantibody-Associated Vasculitis
Authors: Hasegawa Midori , Nishii Chikako , Ohashi Atsushi , Tomita Makoto , Nakai Shigeru , Murakami Kazutaka , Nabeshima Kunihiro , Fujita Yoshirou , Ishii Junichi , Hiki Yoshiyuki , Sugiyama Satoshi
Published: August 2009

Background/Aims: To clarify the clinical significance of tumor necrosis factor (TNF) receptors in patients with myeloperoxidase (MPO)-anti-neutrophil cytoplasmic autoantibody (ANCA)-associated vasculitis, we evaluated the cell surface expression of TNF receptor 1 (TNFR1) and TNF receptor 2 (TNFR2). Patients and Methods: 43 patients with MPO-ANCA-associated vasculitis, 16 patients with chronic renal failure, 10 patients with sepsis, 15 patients with systemic lupus erythematosus, and 18 healthy controls were enrolled in this study, and the surface expression levels of TNFR1, TNFR2, CD63, and CD64 on granulocytes were assessed. In 21 patients with MPO-ANCA-associated vasculitis, soluble TNFR1 (sTNFR1), soluble TNFR2(sTNFR2), and TNF-α in the serum were also measured. Results: The surface expression levels of TNFR1 and TNFR2 on granulocytes were significantly higher in patients with MPO-ANCA-associated vasculitis than in the healthy controls, and positively correlated with the Birmingham Vasculitis Activity Score (BVAS). The levels of sTNFR1, sTNFR2, and TNF-α in the serum were also significantly higher in patients with MPO-ANCA-associated vasculitis than in the healthy controls. Serum levels of sTNFR1 and sTNFR2 correlated with serum creatinine, while the surface expression of TNFR1 and TNFR2 on the granulocytes did not. There was no significant correlation between the BVAS and CD63 or BVAS and CD64. Conclusion: The surface expression levels of TNFR1 and TNFR2 on granulocytes were upregulated in patients with MPO-ANCA-associated vasculitis and reflected disease activity.
Oxidative Modification of Albumin in Predialysis, Hemodialysis, and Peritoneal Dialysis Patients
Authors: Mitrogianni Zoi , Barbouti Alexandra , Galaris Dimitrios , Siamopoulos Kostas C.
Published: August 2009

Background/Aims: Oxidative damage has been reported to be involved in the pathophysiology of chronic kidney disease (CKD) as well as in the pathogenesis of cardiovascular complications of CKD patients. The aim of the present investigation was to evaluate the levels of plasma carbonyl formation, a sensitive marker of enhanced oxidative stress in predialysis, hemodialysis (HD) and peritoneal dialysis (PD) patients. Methods: Plasma samples from 20 apparently healthy control individuals and 127 CKD (stages 2, 3, 4, HD and PD) patients were evaluated by Western blot analysis for the estimation of the levels of protein carbonyl formation. Results: Albumin represented the main plasma carbonylated protein. Increasing carbonylation of albumin was detected along with the severity of CKD, reaching significance at stages 3 and 4 (p < 0.01, compared to healthy controls). The carbonylation of albumin was even higher in the plasma of HD patients (p < 0.001), while in PD patients it was not statistically significant compared to controls (p = 0.224). Conclusions: The data presented in this work indicate that oxidative stress in CKD patients gradually increased during the development of the disease. This stress is probably intensified during HD, but not in PD subjects.
Disturbances of Gastric Electrical Control Activity after Laparotomic Cholecystectomy Are Related to Interleukin-6 Concentrations
Authors: Maruna P. , Frasko R. , Lindner J.
Published: August 2009

Background: The aim of the present study was to characterize the disturbances of gastric electrical control activity after cholecystectomy and to correlate electrogastrographic (EGG) findings with inflammatory markers. Patients and Methods: 52 adult patients were examined in conjunction with planned laparotomic or laparoscopic cholecystectomy. Gastric myoelectrical activity was recorded with a MicroDigitrapper device using three Ag-AgCl disposable skin electrodes. The dominant frequency was calculated using computerized algorithms. Repeated EGG records were correlated with inflammatory parameters – interleukin-6 (IL-6), tumor necrosis factor-α, procalcitonin, and C-reactive protein. Results: Both basal and stimulated gastric electrical frequency decreased significantly in the postoperative phase measured 6 h after the start of surgery (p < 0.01). Postoperative bradygastria (electrical control activity frequency <2.4 cpm) was relatively more expressed after open cholecystectomy. Among the patients with a laparotomic cholecystectomy, IL-6 maximal postoperative concentrations correlated positively with a prolonged restitution of normal gastric activity. Conclusions: EGG examination showed a high sensitivity in the evaluation of gastric electrical activity after both laparotomic and laparoscopic cholecystectomy. A significant correlation of prolonged bradygastria and IL-6 plasma levels supports the role of the inflammatory milieu in the pathogenesis of impaired gastric electrical activity in the postoperative period.
Leisure Activities in Late Life in Relation to Dementia Risk: Principal Component Analysis
Authors: Paillard-Borg Stéphanie , Fratiglioni Laura , Winblad Bengt , Wang Hui-Xin
Published: August 2009

Aims: To explore the underlying dimensions of a set of interrelated lifestyle factors and test the hypothesis that an active lifestyle may protect against dementia. Methods: The study population consisted of 776 participants aged ≥75 years who were dementia-free at both baseline and the first follow-up examinations. Participation in leisure activities was assessed at baseline of the survey, and principal component analysis was used to identify their underlying factors. Results: During the 9-year follow-up from exposure assessment, 212 subjects developed dementia. Higher factor scores of physical, mental, and social dimensions of an active lifestyle was each related to a lower dementia risk. A significant dose-response association between participation in the diverse dimensions and lower dementia risk was observed. Conclusion: An active lifestyle may protect older people against dementia, and the higher level of engagement the stronger protection.
Effect of Long-Term Consumption of Milk Supplemented with Probiotic Lactobacilli and Fluoride on Dental Caries and General Health in Preschool Children: A Cluster-Randomized Study
Authors: Stecksén-Blicks C. , Sjöström I. , Twetman S.
Published: August 2009

The aim of this study was to evaluate the effect of milk supplemented with probiotic bacteria and fluoride on caries development and general health in preschool children. Children 1–5 years of age (n = 248) attending 14 day care centres with 27 units in northern Sweden entered the study. The centres were randomly assigned to two parallel groups: children in the intervention group were served 150 ml milk supplemented with Lactobacillus rhamnosus LB21 (107 CFU/ml) and 2.5 mg fluoride per litre for lunch while the control group received standard milk. The double-blind intervention lasted for 21 months (weekdays) and data were collected through clinical examinations and questionnaires. The primary outcome was caries increment and secondary outcomes were measures of general health. The dropout rate was 25%. The mean baseline caries experience was 0.5 dmfs in the intervention units and 0.6 in the control units and after 21 months 0.9 and 2.2 (p < 0.05). The number of days with sick leave was similar in both groups but the children of the intervention units displayed 60% fewer days with antibiotic therapy (mean 1.9 vs. 4.7 days) and 50% less days with otitis media (0.5 vs. 1.0) (p > 0.05). In children who had participated during the whole 21-month intervention, fewer days with otitis media were reported (0.4 vs. 1.3 days, p < 0.05). No serious side effects were reported. It is concluded that daily consumption of milk containing probiotic bacteria and fluoride reduced caries in preschool children with a prevented fraction of 75%. Additional beneficial health effects were evident.
2′-Ethoxy-5′-Methoxy-2-(5-Methylthienyl)Chalcone Inhibits Collagen-Induced Protein Tyrosine Phosphorylation and Thromboxane Formation during Platelet Aggregation and Adhesion
Authors: Lo Huey-Ming , Huang Tur-Fu , Lin Chun-Nan , Peng Hui-Chin , Wu Wen-Bin
Published: August 2009

Subendothelial collagen supports platelet adhesion, activation, and thrombus growth at sites of vascular damage. Previous studies have shown that chalcones possess antiplatelet activity, but their mechanism of action is not fully understood. In this study, a recently synthesized chalcone, 2′-ethoxy-5′-methoxy-2-(5-methylthienyl)chalcone (EMMTC), was used to investigate chalcone effects on platelet aggregation and adhesion. We found that EMMTC potently inhibited collagen-induced platelet aggregation with an IC50 of 1.01 μmol/l. In contrast, it did not inhibit thrombin- and ADP-induced platelet aggregation. EMMTC could inhibit arachidonic acid (AA)-induced platelet aggregation and collagen- and AA-induced thromboxane B2 (TXB2) formation, suggesting that cyclooxygenase and/or thromboxane synthase were affected during this process. Moreover, EMMTC suppressed collagen-induced protein tyrosine phosphorylation, including phospholipase C-γ2 (PLC-γ2), phosphatidylinositol-3 kinase (PI-3K), Src, Fyn and LAT. Strikingly, EMMTC also blocked platelet adhesion to immobilized collagen and convulxin (a snake venom-derived protein that activates platelet glycoprotein VI receptor). The attenuation of phosphorylation of focal adhesion kinase (FAK) was observed during adhesion. Taken together, our results presented here demonstrate that the chalcone derivative EMMTC affects collagen-induced protein tyrosine phosphorylation and TXB2 formation and functionally blocks collagen-induced platelet aggregation and adhesion.
Regional Cerebral Blood Volume Ratio on Perfusion MRI on the Growth of Infarct Size in Acute Ischemic Stroke
Authors: Lee Su-Yun , Cha Jae-Kwan , Kang Myong-Jin
Published: August 2009

We hypothesized that the relative regional cerebral blood volume (rCBV) ratio could help predict the risk of infarct growth on follow-up magnetic resonance imaging (MRI) in patients with diffusion perfusion mismatch (DPM) on the time-to-peak (TTP) map. We recruited 60 patients with acute middle cerebral arterial (MCA) infarction who had been evaluated by perfusion MRI within 24 h of initial ischemic events, and assessed the predictive role of the rCBV ratio on infarct growth in patients with DPM. Among 60 patients with acute MCA ischemic stroke, 41 (68.3%) patients had DPM on the initial MRI. Follow-up MRI revealed ischemic lesion enlargement in 19 (31.7%) of these 41 patients. The presence of DPM had no effect on the rate of lesion enlargement. Patients with ischemic lesion growth in follow-up images had a significantly lower rCBV ratio than patients without (0.81 ± 0.22 vs. 1.08 ± 0.20, p < 0.01). In this study, the decreased rCBV ratio on perfusion MRI has a predictive value for the growth of ischemic lesions after acute ischemic stroke with DPM on the TTP map.
Intraspinal Dermoid and Epidermoid Tumors: Report of 18 Cases and Reappraisal of the Literature
Authors: van Aalst J. , Hoekstra F. , Beuls E.A.M. , Cornips E.M.J. , Weber J.W. , Sival D.A. , Creytens D.H.K.V. , Vles J.S.H.
Published: August 2009

Intraspinal dermoid and epidermoid tumors are two histopathological subtypes of cutaneous inclusion tumors of the spine. This classification is based on obsolete embryological knowledge. In fact, according to current embryology, both tumor types consist of ectodermal derivatives. Therefore, we hypothesized that dermoid and epidermoid tumors do not differ in clinical practice. To explore this hypothesis, we studied the clinical, radiological and intraoperative findings of 18 patients, and related these findings to the histopathological characteristics of the tumor. No differences were found between dermoid and epidermoid tumors regarding clinical presentation, radiological examination and outcome, while intraoperative diagnosis by the surgeon correlated with the histopathological diagnosis in only 8 of 18 cases. Therefore, the histopathological difference between intraspinal dermoid and epidermoid tumors is not important in clinical practice and should be avoided. A new nomenclature is proposed in which both tumor types are referred to as ‘spinal cutaneous inclusion tumors’.
Triple Lumbar Dermal Sinuses: Unusual Presentation of a Typically Solitary Midline Lesion
Authors: Ansari Saeed , Andrabi Yasir , El Khashab Mostafa , Bateni Farzad , Dadmehr Majid , Iskandar Bermans J. , Nejat Farideh
Published: August 2009

Background: Congenital dermal sinus is a type of closed spinal dysraphism caused by incomplete disjunction. The lesion occurs everywhere along neuraxis predominantly as a solitary midline sinus in the lumbosacral area. Case History: The authors report on a 10-month-old girl with triple congenital dermal sinuses associated with dermoid tumors and intradural extension of one of them. Discussion: Triple dermal sinuses of the lumbosacral area in a single patient have never been described previously. Incomplete disjunction is suggested to cause this abnormality thereafter the orifice may be divided to three sinuses and some mesenchymal tissues separated the two proximal sinuses to a paramedian location.
Completely Intraosseous Atretic Meningocele
Authors: Davidson Laurence , Gonzalez-Gomez Ignacio , McComb J. Gordon
Published: August 2009

Atretic meningoceles are infrequent congenital malformations resulting from the herniation of meninges through a skull defect. All prior reported cases have had a subcutaneous component with or without an underlying osseous defect. We report the unusual case of a completely intraosseous atretic meningocele.
Articulatory Agility in Cognitive Decline
Authors: Östberg Per , Bogdanović Nenad , Wahlund Lars-Olof
Published: August 2009

Objective: Motor speech disorders are believed to be uncommon in early Alzheimer’s disease (AD). However, data from maximum performance tests of motor speech function in AD and related disorders are virtually nonexistent. The aim of this study was to make such data available. Materials and Methods: Sequential speech motion rate was analyzed in 236 memory clinic patients with different levels of cognitive functioning. Results: Sequential speech motion rate was moderately but significantly decreased in mild dementia in AD. About 10% of AD and mild cognitive impairment cases had markedly decreased rates. Rates were strongly reduced in progressive nonfluent aphasia, whereas semantic dementia did not differ from subjective cognitive impairment. Frontotemporal dementia had lower rates than AD. Conclusions: A proportion of patients with cognitive decline has markedly reduced articulatory agility. The cause of this reduction in some patients with mild cognitive impairment and mild AD is unknown. Semantic dementia is not associated with impaired articulatory agility.