bluuebunny/crossref_metadata_2025 huggingface dataset:
GLOBAL ANALYSIS OF A HARVESTED PREDATOR–PREY MODEL INCORPORATING A CONSTANT PREY REFUGE
Authors: CHEN LIUJUAN , CHEN FENGDE
Published: June 2010

A predator–prey model with Holling type II functional response incorporating a constant prey refuge and independent harvesting in either species is investigated. Some sufficient conditions of the instability and stability properties to the equilibria and the existence and uniqueness to limit cycles for the model are obtained. We also show that influence of prey refuge and harvesting efforts on equilibrium density values. One of the surprising finding is that for fixed prey refuge, harvesting has no influence on the final density of the prey species, while the density of predator species is decreasing with the increasing of harvesting effort on prey species and the fixation of harvesting effort on predator species. Numerical simulations are carried out to illustrate the obtained results and the dependence of the dynamic behavior on the harvesting efforts or prey refuge.
DIFFUSIVE INSTABILITY AND TRAVELING WAVES IN A MANGROVE ECOSYSTEM FOOD-CHAIN MODEL WITH DELAYED DETRITUS RECYCLING
Authors: MUKHOPADHYAY B. , BERA ASHOKE , BHATTACHARYYA R.
Published: June 2010

In this paper, a food-chain model in a mangrove ecosystem with detritus recycling is analyzed. From the stability analysis of the delayed homogeneous system, an interval for the parameter representing detritus-detritivores interaction rate is obtained that imparts stability to the system around the coexistent state. Next, we have studied the model in a nonhomogeneous environment. The analysis revealed the existence of a subinterval of the above mentioned interval such that when the above interaction-rate lies within this interval, the system will undergo diffusion driven instability. Finally, we show the existence of travelling wave solutions for the said ecosystem. Numerical simulations are carried out to augment analytical results.
PERISTALTIC FLOW OF A COMPRESSIBLE NON-NEWTONIAN MAXWELLIAN FLUID THROUGH POROUS MEDIUM IN A TUBE
Authors: ELDESOKY ISLAM M. , MOUSA A. A.
Published: June 2010

This work is concerned with the peristaltic transport of a Newtonian and non-Newtonian Maxwellian fluid in an axisymetric cylindrical tube filled with a homogenous porous medium, in which the flow is induced by traveling transversal waves on the tube wall. Like in peristaltic pumping, the traveling transversal waves induce a net flow of the liquid inside the tube. The viscosity as well as the compressibility of the liquid is taken into account. Modified Darcy's law has been used to model the governing equation. The present theoretical model may be considered as mathematical representation to the case of gall bladder and bile duct with stones and dynamics of blood flow in living creatures. The Navier–Stokes equations for an axisymmetric cylindrical tube are solved by means of a perturbation analysis, in which the ratio of the wave amplitude to the radius of the tube is small parameter. In the second order approximation, a net flow induced by the traveling wave is calculated for various values of the compressibility of the liquid, relaxation time and the permeability parameter of porous medium. The calculations disclose that the compressibility of the liquid, the permeability parameter of porous medium and non-Newtonian effects in presence of peristaltic transport have a strong influence of the net flow rate. Finally, the graphical results are reported and discussed for various values of the physical parameters of interest.
Absence of macrophage inflammatory protein-1α does not impact macrophage accumulation in adipose tissue of diet-induced obese mice
Authors: Surmi Bonnie K. , Webb Corey D. , Ristau Alexander C. , Hasty Alyssa H.
Published: June 2010

Macrophages and T-lymphocytes are known to accumulate in the white adipose tissue (WAT) of obese mice and humans, but the factors that cause this infiltration are not yet determined. Chemokines, which attract leukocytes to inflammatory sites, are candidates for this process. Macrophage inflammatory protein-1α (MIP-1α) expression is significantly elevated in WAT of obese mice and humans and positively correlates with fasting plasma insulin, but its potential role in leukocyte recruitment to WAT is unknown. MIP-1α-deficient, heterozygous, and wild-type mice were fed a Western diet (WD) for 16 wk. Plasma lipids, adipose tissue mass, energy expenditure, food intake, liver triglyceride content, and inflammatory cytokine expression were not different among genotypes. Gene expression of macrophage markers F4/80 and CD68, as well as T-lymphocyte marker CD3ε was increased in perigonadal WAT of obese WD-fed mice but was not influenced by MIP-1α expression level. Immunohistochemical analysis of WAT also showed no effect of MIP-1α on macrophage content. Two related chemokines, MIP-1β and RANTES, had reduced expression in obese male MIP-1α-deficient mice compared with wild-type controls ( P ≤ 0.05). In mice fed the WD for 6 wk, WAT macrophage content was unchanged; however, CD8+ T-lymphocytes accumulated to a lesser extent in the MIP-1α-null mice. Therefore, expression of MIP-1α, as well as that of MIP-1β and RANTES, increases as a consequence of weight gain, but these chemokines may not be required for the recruitment of monocytes to WAT during diet-induced obesity in mice and may impact T-lymphocyte recruitment only at early time points after WD feeding.
Skeletal muscle inflammation is not responsible for the rapid impairment in adiponectin response with high-fat feeding in rats
Authors: Mullen Kerry L. , Tishinsky Justine M. , Robinson Lindsay E. , Dyck David J.
Published: June 2010

Adiponectin (Ad) is an insulin-sensitizing adipokine known to stimulate fatty acid (FA) oxidation in skeletal muscle. Skeletal muscle can become resistant to Ad very rapidly, after only 3 days of high saturated fat feeding in rats. Whether the same occurs following a high polyunsaturated fat diet is unknown. Obesity, insulin resistance, and hyperlipidemia are recognized as low-grade inflammatory diseases; therefore, we hypothesized that high-fat feeding induces inflammation, which interferes with Ad action at skeletal muscle. To this end, rats were placed into one of three dietary groups, control (CON, 10% kcal from fat), high saturated (SAT), or high polyunsaturated (PUFA) fat (60% kcal from fat) for 3 days to determine whether Ad resistance develops. Half of the animals from each group were further supplemented with aspirin, a common anti-inflammatory drug. Ad stimulated FA metabolism, Ad signaling intermediates [AdipoR1, APPL1, LKB1, AMPK, and acetyl-CoA carboxylase (ACC)], and inflammatory proteins [Toll-like receptor (TLR4), IKKα/β, IκBα, NF-κB, suppressor of cytokine signaling-3 (SOCS3), and JNK] were measured in soleus muscle. Three days of SAT feeding induced Ad resistance in soleus muscle, assessed as an inability of Ad to phosphorylate ACC and increase FA oxidation. In PUFA-fed animals, Ad-stimulated FA oxidation and ACC phosphorylation to the same degree as CON animals (FA oxidation: +35%, +41%; pACC +29%, +19%; CON, PUFA, P < 0.05). However, neither SAT nor PUFA feeding for 3 days induced skeletal muscle inflammation. Surprisingly, aspirin prevented Ad-stimulated increases in FA oxidation. In conclusion, FA type is critical in the development of Ad resistance, but this does not appear to be mediated by inflammation.
Transcriptome responses to heat stress in the nucleated red blood cells of the rainbow trout (Oncorhynchus mykiss)
Authors: Lewis Johanne M. , Hori Tiago S. , Rise Matthew L. , Walsh Patrick J. , Currie Suzanne
Published: June 2010

The retention of a nucleus in the mature state of fish red blood cells (RBCs) and the ability to easily collect and manipulate blood in nonterminal experiments make blood an ideal tissue on which to study the cellular stress response in fish. Through the use of the cGRASP 16K salmonid microarray, we investigated differences in RBC global gene transcription in fish held under control conditions (11°C) and exposed to heat stress (1 h at 25°C followed by recovery at 11°C). Repeated blood sampling (via a dorsal aorta cannula) enables us to examine the individual stress response over time. Samples were taken preheat stress (representing individual control) and at 4 and 24 h postheat stress (representing early and late transcriptional regulation). Approximately 3,000 microarray features had signal above threshold when hybridized with RBC RNA-derived targets, and cannulation did not have a detectable effect on RBC mRNA expression at the investigated time points. Genes involved in the stress response, immune response, and apoptosis were among those showing the highest dysregulation during both early and late transcriptional regulation. Additionally, genes related to the differentiation and development of blood cells were transcriptionally upregulated at the 24 h time point. This study provides a broader understanding of the mechanisms underpinning the stress response in fish and the discovery of novel genes that are regulated in a stress specific manner. Moreover, salmonid transcripts that are consistently dysregulated in blood in response to heat stress are potential candidates of nonlethal biomarkers of exposure to this particular stressor.
Histochemical Evaluation of Human Prostatic Tissues withCratylia mollisSeed Lectin
Authors: de Lima Amanda L. R. , Cavalcanti Carmelita C. B. , Silva Mariana C. C. , Paiva Patrícia M. G. , Coelho Luana C. B. B. , Beltrão Eduardo I. C. , Correia Maria T. dos S.
Published: June 2010

Lectins, proteins which selectively recognize carbohydrates, have been used in histochemistry for the evaluation of changes in glycosylation in processes of cellular differentiation and/or dedifferentiation.Cratylia mollisseed lectins (Cramoll 1,4 and Cramoll 3), conjugated to horseradish peroxidase, were used as histochemical probes in human prostate tissues: normal (NP), hyperplasia (BPH), and prostate carcinoma (PCa). The staining pattern of Con-A and Cramoll 1,4 in BPH was more intense than in NP. These lectins also showed staining differences between BPH and PCa; the latter showing decreased staining intensity with an increased degree of malignancy. PNA and Cramoll 3 stained epithelial cells similarly in all diagnoses although they did present intense staining of PCa glands lumen. Corpora amylacea were not differentially recognized by any of the lectins. Cramoll 1,4 and Cramoll 3 seed lectins present themselves as candidates for histochemical probes for prostate pathologies when compared to commercial lectins such as Con-A and PNA.
The Battle between Virus and Host: Modulation of Toll-Like Receptor Signaling Pathways by Virus Infection
Authors: Yokota Shin-ichi , Okabayashi Tamaki , Fujii Nobuhiro
Published: June 2010

In order to establish an infection, viruses need to either suppress or escape from host immune defense systems. Recent immunological research has focused on innate immunity as the first line of host defense, especially pattern recognition molecules such as Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), and NOD-like receptors (NLRs). Various microbial components are recognized by their vague and common molecular shapes so-called, pathogen-associated molecular patterns (PAMPs). PAMPs induce inflammatory reactions mediated by the activation of the transcription factor, NF-κB, and by interferons, which lead to an antiviral immune response. Viruses have the capacity to suppress or escape from this pattern recognition molecule-mediated antimicrobial response in various ways. In this paper, we review the various strategies used by viruses to modulate the pattern recognition molecule-mediated innate immune response.
Circulating Endothelial Microparticles in Diabetes Mellitus
Authors: Tramontano A. F. , Lyubarova R. , Tsiakos J. , Palaia T. , DeLeon J. R. , Ragolia L.
Published: June 2010

Background. Endothelial Microparticles (EMPs) are small vesicles shed from activated or apoptotic endothelial cells and involved in cellular cross-talk. Whether EMP immunophenotypes vary according to stimulus in Diabetes Mellitus (DM) is not known. We studied the cellular adhesion molecule (CAM) profile of circulating EMPs in patients with and without Diabetes Mellitus type 2, who were undergoing elective cardiac catheterization.Methods and Results. EMPs were analyzed by flow cytometry. The absolute median number of EMPs (EMPs/L) specific for CD31, CD105, and CD106 was significantly increased in the DM population. The ratio of CD62E/CD31 EMP populations reflected an apoptotic process.Conclusion. Circulating CD31+, CD105+, and CD106+ EMPs were significantly elevated in patients with DM. EMPs were the only independent predictors of DM in our study cohort. In addition, the EMP immunophenotype reflected an apoptotic process. Circulating EMPs may provide new options for risk assessment.
Removal of Organic Matter from Landfill Leachate by Advanced Oxidation Processes: A Review
Authors: Li Wei , Zhou Qixing , Hua Tao
Published: June 2010

In most countries, sanitary landfill is nowadays the most common way to eliminate municipal solid wastes (MSWs). However, sanitary landfill generates large quantity of heavily polluted leachate, which can induce ecological risk and potential hazards towards public health and ecosystems. The application of advanced oxidation processes (AOPs) including ozone-based oxidation, Fenton oxidation, electrochemical oxidation, and other AOPs to treatment of landfill leachate was reviewed. The treatment efficiency in term of chemical oxygen demand (COD) of various AOPs was presented. Advantages and drawbacks of various AOPs were discussed. Among the AOPs reviewed, Fenton process should be the best choice, not only because it can achieve about 49~89% of COD removal with COD ranging from 837 to 8894 mg/L, but also because the process is cost-effective and simple in technological aspect, there is no mass transfer limitation (homogeneous nature) and both iron and hydrogen peroxide are nontoxic.
Stabilization of Curcumin by Complexation with Divalent Cations in Glycerol/Water System
Authors: Zebib Bachar , Mouloungui Zéphirin , Noirot Virginie
Published: June 2010

The purpose of present study was to stabilize curcumin food pigment by its complexation with divalent ions like (Zn2+, Cu2+, Mg2+, Se2+), in “green media” and evaluate its stability in vitro compared to curcumin alone. The curcumin complexes were prepared by mechanical mixture of curcumin and sulfate salts of each metal (metal : curcumin 1/1mol) into unconventional and nontoxic glycerol/water solvent. Two stoichiometry of complex were obtained, 1 : 1 and 1 : 2 (metal/curcumin), respectively. On evaluation of in vitro stability, all complexes were found to provide a higher stability from curcumin alone.
Corneal Deposit of Ciprofloxacin after Laser Assisted Subepithelial Keratomileusis Procedure: A Case Report
Authors: De Benedetti Giacomo , Brancaccio Andrea
Published: June 2010

Purpose. To report one case of corneal antibiotic deposition after ciprofloxacin administration in Laser Assisted Subepithelial Keratomileusis (LASEK).Methods. One case of post-LASEK treatment resulted in corneal precipitates and poor wound healing. Debris was analyzed with dark field microscopy and placed on a blood-agar plate seeded with a susceptible stain ofStaphylococcus aureus(ATCC 29213).Results. The alterations resolved with discontinuation of ciprofloxacin treatment, although some residual deposits persisted subepithelially for 6 months. Analysis of precipitates revealed polydisperse crystalline needles of 183 m average length ( m) and the excised precipitate demonstrated a zone of inhibition.Conclusions. Fluoroquinolone antibiotic drops have been used extensively in postsurgical treatment of refractive surgery. Corneal precipitates have been previously reported in the literature, but up to now nothing has been documented after LASEK. Polypharmacy during refractive surgery may impair epithelialisation, and clinical management should reduce toxic environment and promote ocular surface stability when performing surface ablations.
Antarctic 20th Century Accumulation Changes Based on Regional Climate Model Simulations
Authors: Dethloff Klaus , Glushak Ksenia , Rinke Annette , Handorf Dörthe
Published: June 2010

The regional climate model HIRHAM has been applied to Antarctica driven at the lateral and lower boundaries by European Reanalysis data ERA‐40 for the period 1958–1998. Simulations over 4 decades, carried out with a horizontal resolution of 50 km, deliver a realistic simulation of the Antarctic atmospheric circulation, synoptic‐scale pressure systems, and the spatial distribution of precipitation minus sublimation (P‐E) structures. The simulated P‐E pattern is in qualitative agreement with glaciological estimates. The estimated (P‐E) trends demonstrate surfacemass accumulation increase at the West Antarctic coasts and reductions in parts of East Antarctica.
The influence of the Antarctic Oscillation (AAO) on the near‐surface climate and the surface mass accumulation over Antarctica have been
investigated on the basis of ERA‐40 data and HIRHAM simulations. It is shown that the regional accumulation changes are largely driven by changes in the transient activity around the Antarctic coasts due to the varying AAO phases. During positive AAO, more transient pressure systems travelling towards the continent, and Western Antarctica and parts of South‐Eastern Antarctica gain more precipitation and mass. Over central Antarctica the prevailing anticyclone causes a strengthening of polar desertification connected with a reduced surface mass balance in the northern part of East Antarctica.
Enhancement of Human Prolactin Synthesis by Sodium Butyrate Addition to Serum-Free CHO Cell Culture
Authors: Rodrigues Goulart Herbert , Arthuso Fernanda dos Santos , Capone Marcos Vinicius Nucci , Oliveira Taís Lima de , Bartolini Paolo , Soares Carlos Roberto Jorge
Published: June 2010

Sodium butyrate (NaBu) has been used as a productivity enhancer for the synthesis of recombinant proteins in Chinese hamster ovary (CHO) cells. Thus, the influence of NaBu on the production of recombinant human prolactin (hPRL) from CHO cells was investigated for the first time. CHO cell cultures were submitted to a treatment with different concentrations of NaBu (0.25 to 4 mM). Quantitative and qualitative analyses by reverse-phase high-performance liquid chromatography (RP-HPLC) and Western blot or SDS-PAGE, carried out directly on CHO-conditioned medium, showed that the highest hPRL expression was obtained with 1 mM NaBu. In vitro biological assays based on noble rat lymphoma (Nb2) and mouse pro-B lymphoma (Ba/F3-LLP) cells were carried out on purified hPRL. Its bioactivity in the presence of NaBu was not apparently different from that of the First International Reference Reagent of recombinant hPRL (WHO 97/714). Our results show that NaBu increased the synthesis of recombinant hPRL in CHO cells, apparently without compromising either its structure or function.
New Components for Dye-Sensitized Solar Cells
Authors: Caramori Stefano , Cristino Vito , Boaretto Rita , Argazzi Roberto , Bignozzi Carlo Alberto , Di Carlo Aldo
Published: June 2010

Dye-Sensitized Solar Cells (DSSCs) are among the most promising solar energy conversion devices of new generation, since coupling ease of fabrication and low cost offer the possibility of building integration in photovoltaic windows and facades. Although in their earliest configuration these systems are close to commercialization, fundamental studies are still required for developing new molecules and materials with more desirable properties as well as improving our understanding of the fundamental processes at the basis of the functioning of photoactive heterogeneous interfaces. In this contribution, some recent advances, made in the effort of improving DSSC devices by finding alternative materials and configurations, are reviewed.
The Relationship between Mold Exposure and Allergic Response in Post-Katrina New Orleans
Authors: Rabito Felicia A. , Perry Sara , Davis W. Edward , Yau C. Lillian , Levetin Estelle
Published: June 2010

Objectives. The study's objective was to examine the relation between mold/dampness exposure and mold sensitization among residents of Greater New Orleans following Hurricane Katrina. Methods. Patients were recruited from the Allergy Clinic of a major medical facility. Any patient receiving a skin prick test for one of 24 molds between December 1, 2005 and December 31, 2008 was eligible for the study. Exposure was assessed using standardized questionnaires. Positive mold reactivity was defined as a wheal diameter >3 mm to any mold genera. Results. Approximately 57% of participants tested positive to any indoor allergen, 10% to any mold. Over half of respondents had significant home damage, 34% reported dampness/mold in their home, half engaged in renovation, and one-third lived in a home undergoing renovation. Despite extensive exposure, and multiple measures of exposure, we found no relationship between mold/dampness exposure and sensitivity to mold allergens. Conclusions. These results along with results of earlier research indicate no excess risk of adverse respiratory effects for residents living in New Orleans after the devastation of Hurricane Katrina.
Life-Threatening Adenovirus Infections in the Setting of the Immunocompromised Allogeneic Stem Cell Transplant Patients
Authors: Fowler Cedar J. , Dunlap Jennifer , Troyer Darcy , Stenzel Peter , Epner Elliot , Maziarz Richard T.
Published: June 2010

A single institution case series of adenovirus infections after allogeneic hematopoietic stem cell transplantation is presented to highlight the consideration for adenovirus infections as an etiology in patients with rapid hepatic or other sudden organ deterioration in the setting of apparent GVHD stabilization. The series also highlights that survival is limited with these infections often due in part to concomitant opportunistic infections. In addition, the pathophysiological events, such as GVHD and hepatic dysfunction, may complicate the clinical picture and delay therapy of an opportunistic infection. This is particularly true for adenoviral infections as they also have a distinct clinical picture in immunocompromised patients when compared to immune competent patients. Adenovirus infections also have the additional challenge that its treatment, cidofovir, has associated toxicities that can delay its administration. Recent developments has yielded an assay that can be used in the early detection and for serial determinations of adenovirus in patients with advanced GVHD, as well as a new therapeutic agent currently undergoing clinical trials.
The Resistance to Freeze-Drying and to Storage Was Determined as the Cellular Ability to Recover Its Survival Rate and Acidification Activity
Authors: Coulibaly Ibourahema , Dubois-Dauphin Robin , Destain Jacqueline , Fauconnier Marie-Laure , Lognay Georges , Thonart Philippe
Published: June 2010

The protective effects of the fatty acid composition and membrane action of the acidification activity of two strains ofLactobacilluskept at20∘Cwere studied. The addition of sorbitol, monosodium glutamate and glycerol during storage is causing the decline of acidification and increased concentrations of unsaturated fatty acids observed in both strains. The addition of sorbitol and monosodium glutamate does not alter the fatty acid composition, whatever the strain, but increases the resistance to freeze-drying ofL. plantarumCWBI-B1419 and improves survival during storage. The addition of these preservatives and decreased activity of acidification improves the ratio unsaturated. These results indicate that the survival during storage and freeze-drying resistance are closely related to the composition of membrane fatty acids. This behaviour can be interpreted as an adaptation ofL. plantarumB1419-CWBI supplemented by cryoprotectant additives such as sorbitol or monosodium glutamate sorbitol and monosodium glutamate as an additive.L. plantarumCWBI-B1419 presents a greater adaptation to culture conditions thanL. paracaseissp.paracaseiLMG9192T.
Erythrocyte and Reticulocyte Indices on the LH 750 as Potential Markers of Functional Iron Deficiency
Authors: Urrechaga Eloísa , Borque Luís , Escanero Jesús F.
Published: June 2010

Reticulocyte hemoglobin content (CHr) and percentage of hypochromic cells (%Hypo) are restricted to the Siemens analysers. The aims of the study were to investigate the correlation of Red cells size factor (RSf) and low Hemoglobin density (LHD%), reported by Beckman-Coulter analysers, with CHr and %Hypo in the assessment of iron status in the presence of inflammation. 381 samples were run on both LH 750 (Beckman-Coulter) and Advia 2120 (Siemens) analysers. Correlation between parameters were calculated and the diagnostic performance of the new parameters was assessed.Results. Correlation between RSf and CHr, . ROC curve analysis for RSf in the diagnosis of iron restricted erythropoiesis defined as CH pg: AUC 0.983; Cutoff 91.1%; Sensitivity 98.8%; Specificity 89.6% Correlation between LHD% and %Hypo, . ROC curve analysis for LHD% in the diagnosis of iron deficiency defined by %Hypo %: AUC 0.954; Cut off 6.0%; Sensitivity 96.6%; Specificity 83.2%Conclusions. RSf and LHD% could be reliable parameters for the study of iron metabolism status.
Perinatal S100B Protein Assessment in Human Unconventional Biological Fluids: A Minireview and New Perspectives
Authors: Gazzolo Diego , Michetti Fabrizio
Published: June 2010

Growing evidence is now available on the use of S100B protein as a valuable marker of brain damage and its role as a neurotrophic factor. Bearing in mind, among different S100B protein properties that are still being investigated, the possibility of measuring this protein in different biological fluids renders it suitable for use in several disciplines. This is the case with perinatal medicine where even more noninvasive techniques are particularly desirable in order to ensure the minimal handling diagnostic and therapeutic strategies. In this setting, the present minireview reports data on the presence and the usefulness of S100B protein as brain damage marker and as a neurotrophic factor in the so-called unconventional biological fluids such as saliva and human milk, respectively. Results offer new possibilities for the use of S100B in perinatal medicine as a key-protein for the investigations focusing on central nervous system development and damage.
PIK3CA Mutations in In situ and Invasive Breast Carcinomas
Authors: Miron Alexander , Varadi Maria , Carrasco Daniel , Li Hailun , Luongo Lauren , Kim Hee Jung , Park So Yeon , Cho Eun Yoon , Lewis Gretchen , Kehoe Sarah , Iglehart J. Dirk , Dillon Deborah , Allred D. Craig , Macconaill Laura , Gelman Rebecca , Polyak Kornelia
Published: June 2010

The PIK3 signaling pathway has been identified as one of the most important and most frequently mutated pathways in breast cancer. Somatic mutations in the catalytic subunit of PIK3CA have been found in a significant fraction of breast carcinomas, and it has been proposed that mutant PIK3CA plays a role in tumor initiation. However, the majority of primary human tumors analyzed for genetic alterations in PIK3CA have been invasive breast carcinomas and the frequency of PIK3CA mutations in preinvasive lesions has not been explored. To investigate this, we sequenced exons 9 and 20 of PIK3CA in pure ductal carcinoma in situ (DCIS), DCIS adjacent to invasive carcinoma, and invasive ductal breast carcinomas. In a subset of cases, both in situ and invasive areas were analyzed from the same tumor. We found that the frequency of PIK3CA mutations was essentially the same (∼30%) in all three histologic groups. In some cases, in situ and invasive areas of the same tumor were discordant for PIK3CA status, and in two cases in which multiple invasive and adjacent in situ areas within the same tumor were analyzed independently, we detected intratumor heterogeneity for PIK3CA mutations. Our results suggest that mutation of PIK3CA is an early event in breast cancer that is more likely to play a role in breast tumor initiation than in invasive progression, although a potential role for exon 9 mutations in the progression of a subset of DCIS cases cannot be excluded. Cancer Res; 70(14); 5674–8. ©2010 AACR.
Hypoxic Conversion of SMAD7 Function from an Inhibitor into a Promoter of Cell Invasion
Authors: Heikkinen Pekka T. , Nummela Marika , Jokilehto Terhi , Grenman Reidar , Kähäri Veli-Matti , Jaakkola Panu M.
Published: June 2010

Smad7 is an inhibitor of the transforming growth factor-β–activated signaling pathway. Under well-oxygenated conditions, Smad7 is a potent inhibitor of carcinoma cell invasion. Paradoxically, however, the expression of Smad7 is upregulated across several cancers and may promote cancer progression. Hypoxia, which is frequently met in solid tumors, is an enhancer of carcinoma cell invasion and cancer progression. Here, we report that hypoxia activates the expression of Smad7 in a hypoxia-inducible factor– and von Hippel-Lindau protein–dependent manner. As expected, in normoxia, the forced expression of Smad7 inhibited carcinoma cell invasion. In contrast with the normoxic condition, the inhibitory effect of Smad7 was lost under hypoxia. The block in carcinoma cell invasion by forced expression of Smad7 was released by hypoxia in two invasive carcinoma cell lines. Moreover, the noninvasive HaCaT keratinocytes become invasive upon simultaneous hypoxia and transforming growth factor-β stimulus. The hypoxia-activated invasion was attenuated by inhibiting Smad7 expression by short interfering RNA. Finally, the increased Smad7 expression in human carcinomas correlated with hypoxic gene expression. The data provide evidence that hypoxia could convert Smad7 function from an invasion inhibitor into an activator of invasion. Furthermore, they might shed light as to why increased Smad7 expression is detected in cancers. Cancer Res; 70(14); 5984–93. ©2010 AACR.
Phosphorylation and Activation of Cell Division Cycle Associated 5 by Mitogen-Activated Protein Kinase Play a Crucial Role in Human Lung Carcinogenesis
Authors: Nguyen Minh-Hue , Koinuma Junkichi , Ueda Koji , Ito Tomoo , Tsuchiya Eiju , Nakamura Yusuke , Daigo Yataro
Published: June 2010

We analyzed the gene expression profiles of clinical lung carcinomas using a cDNA microarray containing 27,648 genes or expressed sequence tags, and identified CDCA5 (cell division cycle associated 5) to be upregulated in the majority of lung cancers. Tumor tissue microarray analysis of 262 non–small cell lung cancer patients revealed that CDCA5 positivity was an independent prognostic factor for lung cancer patients. Suppression of CDCA5 expression with siRNAs inhibited the growth of lung cancer cells; concordantly, induction of exogenous expression of CDCA5 conferred growth-promoting activity in mammalian cells. We also found that extracellular signal-regulated kinase (ERK) kinase phosphorylated CDCA5 at Ser79 and Ser209 in vivo. Exogenous expression of phospho-mimicking CDCA5 protein whose Ser209 residue was replaced with glutamine acid further enhanced the growth of cancer cells. In addition, functional inhibition of the interaction between CDCA5 and ERK kinase by a cell-permeable peptide corresponding to a 20-amino-acid sequence part of CDCA5, which included the Ser209 phosphorylation site by ERK, significantly reduced phosphorylation of CDCA5 and resulted in growth suppression of lung cancer cells. Our data suggest that transactivation of CDCA5 and its phosphorylation at Ser209 by ERK play an important role in lung cancer proliferation, and that the selective suppression of the ERK-CDCA5 pathway could be a promising strategy for cancer therapy. Cancer Res; 70(13); 5337–47. ©2010 AACR.
The Phosphoinositide 3-Kinase Inhibitor PI-103 Downregulates Choline Kinase α Leading to Phosphocholine and Total Choline Decrease Detected by Magnetic Resonance Spectroscopy
Authors: Al-Saffar Nada M.S. , Jackson L. Elizabeth , Raynaud Florence I. , Clarke Paul A. , Ramírez de Molina Ana , Lacal Juan C. , Workman Paul , Leach Martin O.
Published: June 2010

The phosphoinositide 3-kinase (PI3K) pathway is a major target for cancer drug development. PI-103 is an isoform-selective class I PI3K and mammalian target of rapamycin inhibitor. The aims of this work were as follows: first, to use magnetic resonance spectroscopy (MRS) to identify and develop a robust pharmacodynamic (PD) biomarker for target inhibition and potentially tumor response following PI3K inhibition; second, to evaluate mechanisms underlying the MRS-detected changes. Treatment of human PTEN null PC3 prostate and PIK3CA mutant HCT116 colon carcinoma cells with PI-103 resulted in a concentration- and time-dependent decrease in phosphocholine (PC) and total choline (tCho) levels (P < 0.05) detected by phosphorus (31P)- and proton (1H)-MRS. In contrast, the cytotoxic microtubule inhibitor docetaxel increased glycerophosphocholine and tCho levels in PC3 cells. PI-103–induced MRS changes were associated with alterations in the protein expression levels of regulatory enzymes involved in lipid metabolism, including choline kinase α (ChoKα), fatty acid synthase (FAS), and phosphorylated ATP-citrate lyase (pACL). However, a strong correlation (r2 = 0.9, P = 0.009) was found only between PC concentrations and ChoKα expression but not with FAS or pACL. This study identified inhibition of ChoKα as a major cause of the observed change in PC levels following PI-103 treatment. We also showed the capacity of 1H-MRS, a clinically well-established technique with higher sensitivity and wider applicability compared with 31P-MRS, to assess response to PI-103. Our results show that monitoring the effects of PI3K inhibitors by MRS may provide a noninvasive PD biomarker for PI3K inhibition and potentially of tumor response during early-stage clinical trials with PI3K inhibitors. Cancer Res; 70(13); 5507–17. ©2010 AACR.
Selective BRAFV600E Inhibition Enhances T-Cell Recognition of Melanoma without Affecting Lymphocyte Function
Authors: Boni Andrea , Cogdill Alexandria P. , Dang Ping , Udayakumar Durga , Njauw Ching-Ni Jenny , Sloss Callum M. , Ferrone Cristina R. , Flaherty Keith T. , Lawrence Donald P. , Fisher David E. , Tsao Hensin , Wargo Jennifer A.
Published: June 2010

Targeted therapy against the BRAF/mitogen-activated protein kinase (MAPK) pathway is a promising new therapeutic approach for the treatment of melanoma. Treatment with selective BRAF inhibitors results in a high initial response rate but limited duration of response. To counter this, investigators propose combining this therapy with other targeted agents, addressing the issue of redundancy and signaling through different oncogenic pathways. An alternative approach is combining BRAF/MAPK-targeted agents with immunotherapy. Preliminary evidence suggests that oncogenic BRAF (BRAFV600E) contributes to immune escape and that blocking its activity via MAPK pathway inhibition leads to increased expression of melanocyte differentiation antigens (MDA). Recognition of MDAs is a critical component of the immunologic response to melanoma, and several forms of immunotherapy capitalize on this recognition. Among the various approaches to inhibiting BRAF/MAPK, broad MAPK pathway inhibition may have deleterious effects on T lymphocyte function. Here, we corroborate the role of oncogenic BRAF in immune evasion by melanoma cells through suppression of MDAs. We show that inhibition of the MAPK pathway with MAPK/extracellular signal-regulated kinase kinase (MEK) inhibitors or a specific inhibitor of BRAFV600E in melanoma cell lines and tumor digests results in increased levels of MDAs, which is associated with improved recognition by antigen-specific T lymphocytes. However, treatment with MEK inhibitors impairs T lymphocyte function, whereas T-cell function is preserved after treatment with a specific inhibitor of BRAFV600E. These findings suggest that immune evasion of melanomas mediated by oncogenic BRAF may be reversed by targeted BRAF inhibition without compromising T-cell function. These findings have important implications for combined kinase-targeted therapy plus immunotherapy for melanoma. Cancer Res; 70(13); 5213–9. ©2010 AACR.
Confirmation of Linkage to and Localization of Familial Colon Cancer Risk Haplotype on Chromosome 9q22
Authors: Gray-McGuire Courtney , Guda Kishore , Adrianto Indra , Lin Chee Paul , Natale Leanna , Potter John D. , Newcomb Polly , Poole Elizabeth M. , Ulrich Cornelia M. , Lindor Noralane , Goode Ellen L. , Fridley Brooke L. , Jenkins Robert , Le Marchand Loic , Casey Graham , Haile Robert , Hopper John , Jenkins Mark , Young Joanne , Buchanan Daniel , Gallinger Steve , Adams Mark , Lewis Susan , Willis Joseph , Elston Robert , Markowitz Sanford D. , Wiesner Georgia L.
Published: June 2010

Genetic risk factors are important contributors to the development of colorectal cancer. Following the definition of a linkage signal at 9q22-31, we fine mapped this region in an independent collection of colon cancer families. We used a custom array of single-nucleotide polymorphisms (SNP) densely spaced across the candidate region, performing both single-SNP and moving-window association analyses to identify a colon neoplasia risk haplotype. Through this approach, we isolated the association effect to a five-SNP haplotype centered at 98.15 Mb on chromosome 9q. This haplotype is in strong linkage disequilibrium with the haplotype block containing HABP4 and may be a surrogate for the effect of this CD30 Ki-1 antigen. It is also in close proximity to GALNT12, also recently shown to be altered in colon tumors. We used a predictive modeling algorithm to show the contribution of this risk haplotype and surrounding candidate genes in distinguishing between colon cancer cases and healthy controls. The ability to replicate this finding, the strength of the haplotype association (odds ratio, 3.68), and the accuracy of our prediction model (∼60%) all strongly support the presence of a locus for familial colon cancer on chromosome 9q. Cancer Res; 70(13); 5409–18. ©2010 AACR.
Nuclear Alternate Estrogen Receptor GPR30 Mediates 17β-Estradiol–Induced Gene Expression and Migration in Breast Cancer–Associated Fibroblasts
Authors: Madeo Antonio , Maggiolini Marcello
Published: June 2010

Fibroblasts are the principal cellular component of connective tissue and are associated with cancer cells at all stages of tumor progression. Structural and functional contributions of fibroblasts to the growth, survival, and invasive capacity of cancer cells are beginning to emerge. In breast carcinoma, ∼80% of stromal fibroblasts termed cancer-associated fibroblasts (CAF) are thought to manifest an activated phenotype that promotes cancer cell proliferation tumor growth at metastatic sites similar to the primary tumor. In this report, we show that CAFs respond to physiologic concentrations of 17β-estradiol (E2) by rapidly inducing extracellular signal-regulated kinase phosphorylation and immediate early gene expression, including c-fos and connective tissue growth factor, and cyclin D1. Notably, the E2 response is mediated by the alternate estrogen receptor GPR30, which interfaces with the epidermal growth factor receptor (EGFR) signaling pathway. In particular, E2 stimulates a physical interaction between GPR30 and phosphorylated EGFR, recruiting them to the cyclin D1 gene promoter. Nuclear localization induced by E2 was confirmed by cellular immunofluorescence methods. GPR30 was required for CAF proliferation and migration induced by E2. Our results provide important new mechanistic insights into how CAFs are stimulated by estrogen through a GPR30-mediated nuclear signaling pathway. More generally, they define estrogenic GPR30 signaling as a functionally important component of the tumor microenvironment. Cancer Res; 70(14); 6036–46. ©2010 AACR.
RG7204 (PLX4032), a Selective BRAFV600E Inhibitor, Displays Potent Antitumor Activity in Preclinical Melanoma Models
Authors: Yang Hong , Higgins Brian , Kolinsky Kenneth , Packman Kathryn , Go Zenaida , Iyer Raman , Kolis Stanley , Zhao Sylvia , Lee Richard , Grippo Joseph F. , Schostack Kathleen , Simcox Mary Ellen , Heimbrook David , Bollag Gideon , Su Fei
Published: June 2010

The BRAFV600E mutation is common in several human cancers, especially melanoma. RG7204 (PLX4032) is a small-molecule inhibitor of BRAFV600E kinase activity that is in phase II and phase III clinical testing. Here, we report a preclinical characterization of the antitumor activity of RG7204 using established in vitro and in vivo models of malignant melanoma. RG7204 potently inhibited proliferation and mitogen-activated protein/extracellular signal-regulated kinase (ERK) kinase and ERK phosphorylation in a panel of tumor cell lines, including melanoma cell lines expressing BRAFV600E or other mutant BRAF proteins altered at codon 600. In contrast, RG7204 lacked activity in cell lines that express wild-type BRAF or non-V600 mutations. In several tumor xenograft models of BRAFV600E-expressing melanoma, we found that RG7204 treatment caused partial or complete tumor regressions and improved animal survival, in a dose-dependent manner. There was no toxicity observed in any dose group in any of the in vivo models tested. Our findings offer evidence of the potent antitumor activity of RG7204 against melanomas harboring the mutant BRAFV600E gene. Cancer Res; 70(13); 5518–27. ©2010 AACR.
Targeted Activation of RNA Helicase Retinoic Acid–Inducible Gene-I Induces Proimmunogenic Apoptosis of Human Ovarian Cancer Cells
Authors: Kübler Kirsten , Gehrke Nadine , Riemann Soheila , Böhnert Volker , Zillinger Thomas , Hartmann Evelyn , Pölcher Martin , Rudlowski Christian , Kuhn Walther , Hartmann Gunther , Barchet Winfried
Published: June 2010

Most malignant cells are poorly immunogenic and fail to elicit an effective antitumor immune response. In contrast, viral infections of cells are promptly detected and eliminated by the immune system. Viral recognition critically hinges on cytosolic nucleic acid receptors that include the proinflammatory RNA helicase retinoic acid–inducible gene-I (RIG-I). Here, we show that targeted delivery of RIG-I agonists induced ovarian cancer cells to upregulate HLA class I and to secrete the proinflammatory cytokines CXCL10, CCL5, interleukin-6, tumor necrosis factor-α, and IFN-β. Ovarian cancer cells stimulated via RIG-I became apoptotic and were readily phagocytosed by monocytes and monocyte-derived dendritic cells, which in turn upregulated HLA class I/II and costimulatory molecules and released CXCL10 and IFN-α. Our findings offer proof of principle that mimicking viral infection in ovarian cancer cells triggers an immunogenic form of tumor cell apoptosis that may enhance immunotherapy of ovarian cancer. Cancer Res; 70(13); 5293–304. ©2010 AACR.
Clinical Implications of Gene Dosage and Gene Expression Patterns in Diploid Breast Carcinoma
Authors: Parris Toshima Z. , Danielsson Anna , Nemes Szilárd , Kovács Anikó , Delle Ulla , Fallenius Ghita , Möllerström Elin , Karlsson Per , Helou Khalil
Published: June 2010

Purpose: Deregulation of key cellular pathways is fundamental for the survival and expansion of neoplastic cells. In cancer, regulation of gene transcription can be mediated in a variety of ways. The purpose of this study was to assess the impact of gene dosage on gene expression patterns and the effect of other mechanisms on transcriptional levels, and to associate these genomic changes with clinicopathologic parameters.

Experimental Design: We screened 97 invasive diploid breast tumors for DNA copy number alterations and changes in transcriptional levels using array comparative genomic hybridization and expression microarrays, respectively.

Results: The integrative analysis identified an increase in the overall number of genetic alterations during tumor progression and 15 specific genomic regions with aberrant DNA copy numbers in at least 25% of the patient population, i.e., 1q22, 1q22-q23.1, 1q25.3, 1q32.1, 1q32.1-q32.2, 8q21.2-q21.3, 8q22.3, 8q24.3, and 16p11.2 were recurrently gained, whereas 11q25, 16q21, 16q23.3, and 17p12 were frequently lost (P < 0.01). An examination of the expression patterns of genes mapping within the detected genetic aberrations identified 47 unique genes and 1 Unigene cluster significantly correlated between the DNA and relative mRNA levels. In addition, more malignant tumors with normal gene dosage levels displayed a recurrent overexpression of UBE2C, S100A8, and CBX2, and downregulation of LOC389033, STC2, DNALI1, SCUBE2, NME5, SUSD3, SERPINA11, AZGP1, and PIP.

Conclusions: Taken together, our findings suggest that the dysregulated genes identified here are critical for breast cancer initiation and progression, and could be used as novel therapeutic targets for drug development to complement classical clinicopathologic features. Clin Cancer Res; 16(15); 3860–74. ©2010 AACR.
Validation of the FOUR Score (Spanish Version) in Acute Stroke: An Interobserver Variability Study
Authors: Idrovo Luis , Fuentes Blanca , Medina Josmarlin , Gabaldón Laura , Ruiz-Ares Gerardo , Abenza María José , Aguilar-Amat María José , Martínez-Sánchez Patricia , Rodríguez Luis , Cazorla Rubén , Martínez Marta , Tafur Alfonso , Wijdicks Eelco F.M. , Diez-Tejedor Exuperio
Published: June 2010

Background: Methods to assess impaired consciousness in acute stroke typically include the Glasgow Coma Scale (GCS), but the verbal component has limitations in aphasic or intubated patients. The FOUR (Full Outline of UnResponsiveness) score, a new coma scale, evaluates 4 components: eye and motor responses, brainstem reflexes and respiration. We aimed to study the interobserver variability of the FOUR score in acute stroke patients. Methods: We prospectively enrolled consecutive patients with acute stroke admitted from February to July 2008 to the stroke unit of our Neurology Department. Patients were evaluated by neurology residents and nurses using the FOUR score and the GCS. For both scales, we obtained paired and total weighted kappa values (Kw) and intraclass correlation coefficients (ICC). NIH stroke scale was also recorded on admission. Results: We obtained a total of 75 paired evaluations in 60 patients (41 cerebral infarctions, 15 cerebral hemorrhages and 4 transient ischemic attacks). Thirty-three (55%) patients were alert, 17 (28.3%) drowsy and 10 (16.7%) stuporous or comatose. The overall rater agreement was excellent in the FOUR score (Kw 0.93; 95% CI 0.89–0.97) with an ICC of 0.94 (95% CI 0.91–0.96) and in the GCS (Kw 0.96; 95% CI 0.94–0.98) with an ICC of 0.96 (95% CI 0.93–0.97). A good correlation was found between the FOUR score and the GCS (ρ 0.83; p < 0.01) and between the FOUR score and the NIH stroke scale (ρ –0.78; p < 0.001). Conclusions: The FOUR score is a reliable scale for evaluating the level of consciousness in acute stroke patients, showing a good correlation with the GCS and the NIH stroke scale.