bluuebunny/crossref_metadata_2025 huggingface dataset:
Liquid Cell Transmission Electron Microscopy
Authors: Liao Hong-Gang , Zheng Haimei
Published: May 2016

Liquid cell transmission electron microscopy (TEM) has attracted significant interest in recent years. With nanofabricated liquid cells, it has been possible to image through liquids using TEM with subnanometer resolution, and many previously unseen materials dynamics have been revealed. Liquid cell TEM has been applied to many areas of research, ranging from chemistry to physics, materials science, and biology. So far, topics of study include nanoparticle growth and assembly, electrochemical deposition and lithiation for batteries, tracking and manipulation of nanoparticles, catalysis, and imaging of biological materials. In this article, we first review the development of liquid cell TEM and then highlight progress in various areas of research. In the study of nanoparticle growth, the electron beam can serve both as the illumination source for imaging and as the input energy for reactions. However, many other research topics require the control of electron beam effects to minimize electron beam damage. We discuss efforts to understand electron beam–liquid matter interactions. Finally, we provide a perspective on future challenges and opportunities in liquid cell TEM.
Cladosporium fulvum Effectors: Weapons in the Arms Race with Tomato
Authors: de Wit Pierre J.G.M.
Published: May 2016

In this review, I recount my personal history. My drive to study host-pathogen interactions was to find alternatives for agrochemicals, which was triggered after reading the book “Silent Spring” by Rachel Carson. I reflect on my research at the Laboratory of Phytopathology at Wageningen University, where I have worked for my entire career on the interaction between Cladosporium fulvum and tomato, and related gene-for-gene pathosystems. I describe different methods used to identify and sequence avirulence (Avr) genes from the pathogen and resistance (R) genes from the host. The major genes involved in classical gene-for-gene interactions have now been identified, and breeders can produce plants with multiple R genes providing durable and environmentally safe protection against pathogens. In some cases, this might require the use of genetically modified plants when R genes cannot be introduced by classical breeding.
Cross-Border Migration and Social Inequalities
Authors: Faist Thomas
Published: May 2016

Cross-border migration is a visible reflection of global inequalities. Much literature deals with the link between migration and inequalities indirectly, often through topics such as migration and development or the integration of migrants. Surprisingly, little research addresses directly the role of social inequalities. This gap raises at least two major questions: First, how do social inequalities affect opportunities for cross-border migration for different socioeconomic groups? Second, conversely, how do the outcomes of migration affect social inequalities in global patterns of distribution and in life chances in the countries of emigration and of immigration? Of ultimate interest is whether migration buttresses the dominant forms of social stratification or transforms the distribution of valued goods in a fundamental way. Overall, this review suggests that cross-border migration both constitutes a path to upward social mobility for migrants and tends to reinforce durable inequalities on a deeper level.
Learning Beyond the School Walls: Trends and Implications
Authors: Park Hyunjoon , Buchmann Claudia , Choi Jaesung , Merry Joseph J.
Published: May 2016

Academically-focused learning activities beyond formal schooling are expanding in myriad forms throughout the world. This diverse realm of learning activities includes private supplementary education purchased by families such as private tutoring, online courses, cram schools, and learning center franchises. Some public schools also provide academically oriented after-school programs beyond their formal curricula. This review identifies factors relating to students, families, schools, and educational systems that affect participation in supplementary education. Macro forces are also related to the proliferation of learning activities outside of formal schooling. We discuss implications of this trend for educational stratification as well as challenges it creates for families and formal educational systems. Finally, we suggest promising new avenues for data collection and empirical research.
β-Cell deletion of Nr4a1 and Nr4a3 nuclear receptors impedes mitochondrial respiration and insulin secretion
Authors: Reynolds Merrick S. , Hancock Chad R. , Ray Jason D. , Kener Kyle B. , Draney Carrie , Garland Kevin , Hardman Jeremy , Bikman Benjamin T. , Tessem Jeffery S.
Published: May 2016

β-Cell insulin secretion is dependent on proper mitochondrial function. Various studies have clearly shown that the Nr4a family of orphan nuclear receptors is essential for fuel utilization and mitochondrial function in liver, muscle, and adipose. Previously, we have demonstrated that overexpression of Nr4a1 or Nr4a3 is sufficient to induce proliferation of pancreatic β-cells. In this study, we examined whether Nr4a expression impacts pancreatic β-cell mitochondrial function. Here, we show that β-cell mitochondrial respiration is dependent on the nuclear receptors Nr4a1 and Nr4a3. Mitochondrial respiration in permeabilized cells was significantly decreased in β-cells lacking Nr4a1 or Nr4a3. Furthermore, respiration rates of intact cells deficient for Nr4a1 or Nr4a3 in the presence of 16 mM glucose resulted in decreased glucose mediated oxygen consumption. Consistent with this reduction in respiration, a significant decrease in glucose-stimulated insulin secretion rates is observed with deletion of Nr4a1 or Nr4a3. Interestingly, the changes in respiration and insulin secretion occur without a reduction in mitochondrial content, suggesting decreased mitochondrial function. We establish that knockdown of Nr4a1 and Nr4a3 results in decreased expression of the mitochondrial dehydrogenase subunits Idh3g and Sdhb. We demonstrate that loss of Nr4a1 and Nr4a3 impedes production of ATP and ultimately inhibits glucose-stimulated insulin secretion. These data demonstrate for the first time that the orphan nuclear receptors Nr4a1 and Nr4a3 are critical for β-cell mitochondrial function and insulin secretion.
Quantitative Assessment of the Effects of Oxidants on Antigen‐Antibody Binding In Vitro
Authors: Han Shuang , Wang Guanyu , Xu Naijin , Liu Hui
Published: May 2016

Objective. We quantitatively assessed the influence of oxidants on antigen‐antibody‐binding activity. Methods. We used several immunological detection methods, including precipitation reactions, agglutination reactions, and enzyme immunoassays, to determine antibody activity. The oxidation‐reduction potential was measured in order to determine total serum antioxidant capacity. Results. Certain concentrations of oxidants resulted in significant inhibition of antibody activity but had little influence on total serum antioxidant capacity. Conclusions. Oxidants had a significant influence on interactions between antigen and antibody, but minimal effect on the peptide of the antibody molecule.
Hataedock Treatment Has Preventive Therapeutic Effects in Atopic Dermatitis‐Induced NC/Nga Mice under High‐Fat Diet Conditions
Authors: Cha Ho-Yeol , Ahn Sang-hyun , Cheon Jin-Hong , Park In-Sik , Kim Jin-Tack , Kim Kibong
Published: May 2016

This study investigated the preventive therapeutic effects of Hataedock (HTD) treatment on inflammatory regulation and skin protection in AD‐induced NC/Nga mice under high‐fat diet conditions. Before inducing AD, the extract of Coptidis Rhizoma and Glycyrrhiza uralensis was administered orally to the 3‐week‐old mice. After that, AD‐like skin lesions were induced by applying DNFB. All groups except the control group were fed a high‐fat diet freely. We identified the effects of HTD on morphological changes, cytokine release and the induction of apoptosis through histochemistry, immunohistochemistry, and TUNEL assay. HTD downregulated the levels of IL‐4 and PKC but increased the levels of LXR. HTD also suppressed the mast cell degranulation and release of MMP‐9, Substance P. The levels of TNF‐α, p‐IκB, iNOS, and COX‐2 were also decreased. The upregulation of inflammatory cell’s apoptosis is confirmed by our results as increase of apoptotic body and cleaved caspase‐3 and decrease of Bcl‐2. HTD also reduced edema, angiogenesis, and skin lesion inflammation. Our results indicate HTD suppresses various inflammatory response on AD‐induced mice with obesity through the regulation of Th2 differentiation and the protection of lipid barrier. Therefore, HTD could be used as an alternative and preventive therapeutic approach in the management of AD.
miRNA-375 a Sensor of Glucotoxicity Is Altered in the Serum of Children with Newly Diagnosed Type 1 Diabetes
Authors: Marchand Lucien , Jalabert Audrey , Meugnier Emmanuelle , Van den Hende Kathleen , Fabien Nicole , Nicolino Marc , Madec Anne-Marie , Thivolet Charles , Rome Sophie
Published: May 2016

Background.The use of miRNAs as biomarkers for Type 1 Diabetes (T1D) risk is attractive as T1D is usually diagnosed in front of acute symptoms. As miR-375 is highly expressed in the endocrine pancreas, we postulated that its circulating level might reflect beta cell alterations and might be altered in the blood of T1D patients recently diagnosed.Methods.Sera were obtained from 22 T1D children at onset of the disease, before subcutaneous insulin treatment, and from 10 nondiabetic pediatric controls. MiR-375 seric level was quantified by stem-loop RT-PCR-based assay. MiRNAs regulations in isolated human islets in response to high glucose concentrations were determined by TaqMan Low-Density Array.Results.The abundance of miR-375, among the 410 miRNAs detected in human islets, mirrored its well-established role in rodent islet biology. Upregulated miRNAs targeted genes involved in islet homeostasis and regulation of beta cell mass. Downregulated miRNAs, including miR-375, were involved in pancreas secretion and protein turnover. Seric level of miR-375 was lower in T1D children versus age-matched controls, without any correlations with HbA1c, glycaemia, and number of autoantibodies.Conclusion.Altered circulating level of miR-375 at onset of T1D might be a general biomarker of metabolic alterations and inflammation associated with the disease.
Adsorption of Polyvinylpyrrolidone over the Silica Surface: As Affected by Pretreatment of Adsorbent and Molar Mass of Polymer Adsorbate
Authors: Al-Harbi Laila M. , Kosa Samia A. , Baloch Musa K. , Bhatti Qaisar A. , El-Mossalamy El-Sayed El-Badawey H.
Published: May 2016

The adsorption of polyvinylpyrrolidone over the surface of silica has been investigated. The impact of molar mass of the polymer, pH, and pretreatment temperature of silica particles have been evaluated by means of FTIR spectroscopy and electrophoretic measurements. The silica particles used have narrow particle size distribution. The zeta potential of the aqueous silica suspension was decreased with the increase in pH. The amount of polymer adsorbed was increased with the increase in pretreatment temperature, time, concentration, pH, zeta potential, and molar mass of the polymer. The addition of polymer to the system increased the zeta potential due to adsorption of polymer on the surface of the particles. However, the impact increased with the increase in molecular mass of the polymer. The IR spectra obtained before and after adsorption of polymer concluded that, mostly, hydrogen bonding is responsible for the adsorption phenomena; however, hydrophobic interactions also play a significant role. The mechanism has been investigated and established through FTIR spectroscopy.
Prolactin Rescues Immature B-Cells from Apoptosis Induced by B-Cell Receptor Cross-Linking
Authors: Flores-Fernández Rocio , Blanco-Favela Francisco , Fuentes-Pananá Ezequiel M. , Chávez-Sánchez Luis , Gorocica-Rosete Patricia , Pizaña-Venegas Alberto , Chávez-Rueda Adriana Karina
Published: May 2016

Prolactin has an immunomodulatory effect and has been associated with B-cell-triggered autoimmune diseases, such as systemic lupus erythematosus (SLE). In mice that develop SLE, the PRL receptor is expressed in early bone marrow B-cells, and increased levels of PRL hasten disease manifestations, which are correlated with a reduction in the absolute number of immature B-cells. The aim of this work was to determine the effect of PRL in anin vitrosystem of B-cell tolerance using WEHI-231 cells and immature B-cells from lupus prone MRL/lpr mice. WEHI-231 cells express the long isoform of the PRL receptor, and PRL rescued the cells from cell death by decreasing the apoptosis induced by the cross-linking of the B-cell antigen receptor (BCR) as measured by Annexin V and active caspase-3. This decrease in apoptosis may have been due to the PRL and receptor interaction, which increased the relative expression of antiapoptotic Bcl-xL and decreased the relative expression of proapoptotic Bad. In immature B-cells from MRL/lpr mice, PRL increased the viability and decreased the apoptosis induced by the cross-linking of BCR, which may favor the maturation of self-reactive B-cells and contribute to the onset of disease.
Neurologic Adverse Events Associated with Voriconazole Therapy: Report of Two Pediatric Cases
Authors: Demir Sevliya Öcal , Atici Serkan , Akkoç Gülşen , Yakut Nurhayat , İkizoğlu Nilay Baş , Eralp Ela Erdem , Soysal Ahmet , Bakir Mustafa
Published: May 2016

Although voriconazole, a triazole antifungal, is a safe drug, treatment with this agent is associated with certain adverse events such as hepatic, neurologic, and visual disturbances. The current report presents two cases, one a 9-year-old boy and the other a 17-year-old girl, who experienced neurologic side effects associated with voriconazole therapy. Our aim is to remind readers of the side effects of voriconazole therapy in order to prevent unnecessary investigations especially for psychological and ophthalmologic problems. The first case was a 9-year-old boy with cystic fibrosis and invasive aspergillosis that developed photophobia, altered color sensation, and fearful visual hallucination. The second case was a 17-year-old girl with cystic fibrosis and allergic bronchopulmonary aspergillosis, and she experienced photophobia, fatigue, impaired concentration, and insomnia, when the dose of voriconazole therapy was increased from 12 mg/kg/day to 16 mg/kg/day. The complaints of the two patients disappeared after discontinuation of voriconazole therapy. Our experience in these patients reminded us of the importance of being aware of the neurologic adverse events associated with voriconazole therapy in establishing early diagnosis and initiating prompt treatment. In addition, although serum voriconazole concentration was not measured in the present cases, therapeutic drug monitoring for voriconazole seems to be critically important in preventing neurologic side effects in pediatric patients.
Generalized ESO and Predictive Control Based Robust Autopilot Design
Authors: Panchal Bhavnesh , Talole S. E.
Published: May 2016

A novel continuous time predictive control and generalized extended state observer (GESO) based acceleration tracking pitch autopilot design is proposed for a tail controlled, skid-to-turn tactical missile. As the dynamics of missile are significantly uncertain with mismatched uncertainty, GESO is employed to estimate the state and uncertainty in an integrated manner. The estimates are used to meet the requirement of state and to robustify the output tracking predictive controller designed for nominal system. Closed loop stability for the controller-observer structure is established. An important feature of the proposed design is that it does not require any specific information about the uncertainty. Also the predictive control design yields the feedback control gain and disturbance compensation gain simultaneously. Effectiveness of GESO in estimation of the states and uncertainties and in robustifying the predictive controller in the presence of parametric uncertainties, external disturbances, unmodeled dynamics, and measurement noise is illustrated by simulation.
Phenotypic Plasticity and Selection: Nonexclusive Mechanisms of Adaptation
Authors: Grenier S. , Barre P. , Litrico I.
Published: May 2016

Selection and plasticity are two mechanisms that allow the adaptation of a population to a changing environment. Interaction between these nonexclusive mechanisms must be considered if we are to understand population survival. This review discusses the ways in which plasticity and selection can interact, based on a review of the literature on selection and phenotypic plasticity in the evolution of populations. The link between selection and phenotypic plasticity is analysed at the level of the individual. Plasticity can affect an individual’s response to selection and so may modify the end result of genetic diversity evolution at population level. Genetic diversity increases the ability of populations or communities to adapt to new environmental conditions. Adaptive plasticity increases individual fitness. However this effect must be viewed from the perspective of the costs of plasticity, although these are not easy to estimate. It is becoming necessary to engage in new experimental research to demonstrate the combined effects of selection and plasticity for adaptation and their consequences on the evolution of genetic diversity.
Modeling Alzheimer’s Disease with Induced Pluripotent Stem Cells: Current Challenges and Future Concerns
Authors: Zhang Weiwei , Jiao Bin , Zhou Miaojin , Zhou Tao , Shen Lu
Published: May 2016

Alzheimer’s disease (AD) is the most prevalent type of dementia and its pathology is characterized by deposition of extracellular β‐amyloid plaques, intracellular neurofibrillary tangles, and extensive neuron loss. While only a few familial AD cases are due to mutations in three causative genes (APP, PSEN1, and PSEN2), the ultimate cause behind the rest of the cases, called sporadic AD, remains unknown. Current animal and cellular models of human AD, which are based on the Aβ and tau hypotheses only, partially resemble the familial AD. As a result, there is a pressing need for the development of new models providing insights into the pathological mechanisms of AD and for the discovery of ways to treat or delay the onset of the disease. Recent preclinical research suggests that stem cells can be used to model AD. Indeed, human induced pluripotent stem cells can be differentiated into disease‐relevant cell types that recapitulate the unique genome of a sporadic AD patient or family member. In this review, we will first summarize the current research findings on the genetic and pathological mechanisms of AD. We will then highlight the existing induced pluripotent stem cell models of AD and, lastly, discuss the potential clinical applications in this field.
In VivoNear-Infrared Fluorescence Imaging of Aqueous Humor Outflow Structures
Authors: Zeppa L. , Ambrosone L. , Guerra G. , Fortunato M. , Costagliola C.
Published: May 2016

The aim of this study has been to visualize the aqueous outflow system in patients affected by primary open angle glaucoma. A solution of indocyanine green (ICG) plus high viscosity viscoelastic solution was injected into the Schlemm canal during surgery in 10 glaucomatous patients undergoing canaloplasty. Soon after injection of the dye the borders of the scleral flap were completely stained due to partial reflux caused by the intrachannel resistance; progression of the dye along the Schlemm canal starting from the site of injection was then visualized. The filling of the collector channels was observed only in the patent portions of the Schlemm canal. The only noticeable aqueous veins were located in correspondence of the quadrant in which both the Schlemm canal and the collectors were patent. Lastly, a retrograde filling, of glomerular-shaped structures, deepest to the Schlemm canal was observed in the quadrants where the pathway was functioning. Our findings show that injection of a mixture composed of ICG and viscoelastic solution into the Schlemm canal allows a clear visualization of the functioning portions of the conventional outflow pathway. In addition, a retrograde filling of structures presumably located into the iris was also recorded.Clinical Trial Registration. Our study is registered in ISRCTN registry, number54005880, DOI 10.1186/ISRCTN54005880.
Ambient Air and Hole Transport Layer Free Synthesis: Towards Low Cost CH3NH3PbI3Solar Cells
Authors: Ahmed Muhammad Imran , Saleem Hareema , Khan Ahmed Nawaz , Habib Amir
Published: May 2016

Perovskite absorbers have witnessed a remarkable efficiency increase in last couple of years. To meet the commercialization challenge, reduced cost and improved efficiency are the two critical factors. We report on a hole transport layer free device synthesized under ambient air conditions of high humidity of 50% using TiO2-graphene oxide nanocomposite as electron selective contact. The devices achieved a power conversion efficiency of 5.9%. We introduce a novel synthesis route for TiO2-graphene oxide (GO) composite allowing superior charge transport properties. Incorporation of GO in TiO2allows achieving higher power conversion efficiencies while working under ambient air conditions. Ambient air synthesis with hole transport free architecture has the potential to reduce the cost of this technology leading to commercial viability.
Early Epigenetic Downregulation of microRNA-192 Expression Promotes Pancreatic Cancer Progression
Authors: Botla Sandeep K. , Savant Soniya , Jandaghi Pouria , Bauer Andrea S. , Mücke Oliver , Moskalev Evgeny A. , Neoptolemos John P. , Costello Eithne , Greenhalf William , Scarpa Aldo , Gaida Matthias M. , Büchler Markus W. , Strobel Oliver , Hackert Thilo , Giese Nathalia A. , Augustin Hellmut G. , Hoheisel Jörg D.
Published: May 2016

Pancreatic ductal adenocarcinoma (PDAC) is characterized by very early metastasis, suggesting the hypothesis that metastasis-associated changes may occur prior to actual tumor formation. In this study, we identified miR-192 as an epigenetically regulated suppressor gene with predictive value in this disease. miR-192 was downregulated by promoter methylation in both PDAC and chronic pancreatitis, the latter of which is a major risk factor for the development of PDAC. Functional studies in vitro and in vivo in mouse models of PDAC showed that overexpression of miR-192 was sufficient to reduce cell proliferation and invasion. Mechanistic analyses correlated changes in miR-192 promoter methylation and expression with epithelial–mesenchymal transition. Cell proliferation and invasion were linked to altered expression of the miR-192 target gene SERPINE1 that is encoding the protein plasminogen activator inhibitor-1 (PAI-1), an established regulator of these properties in PDAC cells. Notably, our data suggested that invasive capacity was altered even before neoplastic transformation occurred, as triggered by miR-192 downregulation. Overall, our results highlighted a role for miR-192 in explaining the early metastatic behavior of PDAC and suggested its relevance as a target to develop for early diagnostics and therapy. Cancer Res; 76(14); 4149–59. ©2016 AACR.
Three-Dimensional Breast Cancer Models Mimic Hallmarks of Size-Induced Tumor Progression
Authors: Singh Manjulata , Mukundan Shilpaa , Jaramillo Maria , Oesterreich Steffi , Sant Shilpa
Published: May 2016

Tumor size is strongly correlated with breast cancer metastasis and patient survival. Increased tumor size contributes to hypoxic and metabolic gradients in the solid tumor and to an aggressive tumor phenotype. Thus, it is important to develop three-dimensional (3D) breast tumor models that recapitulate size-induced microenvironmental changes and, consequently, natural tumor progression in real time without the use of artificial culture conditions or gene manipulations. Here, we developed size-controlled multicellular aggregates (“microtumors”) of subtype-specific breast cancer cells by using non-adhesive polyethylene glycol dimethacrylate hydrogel microwells of defined sizes (150–600 μm). These 3D microtumor models faithfully represent size-induced microenvironmental changes, such as hypoxic gradients, cellular heterogeneity, and spatial distribution of necrotic/proliferating cells. These microtumors acquire hallmarks of tumor progression in the same cell lines within 6 days. Of note, large microtumors of hormone receptor–positive cells exhibited an aggressive phenotype characterized by collective cell migration and upregulation of mesenchymal markers at mRNA and protein level, which was not observed in small microtumors. Interestingly, triple-negative breast cancer (TNBC) cell lines did not show size-dependent upregulation of mesenchymal markers. In conclusion, size-controlled microtumor models successfully recapitulated clinically observed positive association between tumor size and aggressive phenotype in hormone receptor–positive breast cancer while maintaining clinically proven poor correlation of tumor size with aggressive phenotype in TNBC. Such clinically relevant 3D models generated under controlled experimental conditions can serve as precise preclinical models to study mechanisms involved in breast tumor progression as well as antitumor drug effects as a function of tumor progression. Cancer Res; 76(13); 3732–43. ©2016 AACR.
Copy Number Alterations in Enzyme-Coding and Cancer-Causing Genes Reprogram Tumor Metabolism
Authors: Sharma Ashwini Kumar , Eils Roland , König Rainer
Published: May 2016

Somatic copy number alterations frequently occur in the cancer genome affecting not only oncogenic or tumor suppressive genes, but also passenger and potential codriver genes. An intrinsic feature resulting from such genomic perturbations is the deregulation in the metabolism of tumor cells. In this study, we have shown that metabolic and cancer-causing genes are unexpectedly often proximally positioned in the chromosome and share loci with coaltered copy numbers across multiple cancers (19 cancer types from The Cancer Genome Atlas). We have developed an analysis pipeline, Identification of Metabolic Cancer Genes (iMetCG), to infer the functional impact on metabolic remodeling from such coamplifications and codeletions and delineate genes driving cancer metabolism from those that are neutral. Using our identified metabolic genes, we were able to classify tumors based on their tissue and developmental origins. These metabolic genes were similar to known cancer genes in terms of their network connectivity, isoform frequency, and evolutionary features. We further validated these identified metabolic genes by (i) using gene essentiality data from several tumor cell lines, (ii) showing that these identified metabolic genes are strong indicators for patient survival, and (iii) observing a significant overlap between our identified metabolic genes and known cancer-metabolic genes. Our analyses revealed a hitherto unknown generic mechanism for large-scale metabolic reprogramming in cancer cells based on linear gene proximities between cancer-causing and -metabolic genes. We have identified 119 new metabolic cancer genes likely to be involved in rewiring cancer cell metabolism. Cancer Res; 76(14); 4058–67. ©2016 AACR.
Mitochondria-Targeted Analogues of Metformin Exhibit Enhanced Antiproliferative and Radiosensitizing Effects in Pancreatic Cancer Cells
Authors: Cheng Gang , Zielonka Jacek , Ouari Olivier , Lopez Marcos , McAllister Donna , Boyle Kathleen , Barrios Christy S. , Weber James J. , Johnson Bryon D. , Hardy Micael , Dwinell Michael B. , Kalyanaraman Balaraman
Published: May 2016

Metformin (Met) is an approved antidiabetic drug currently being explored for repurposing in cancer treatment based on recent evidence of its apparent chemopreventive properties. Met is weakly cationic and targets the mitochondria to induce cytotoxic effects in tumor cells, albeit not very effectively. We hypothesized that increasing its mitochondria-targeting potential by attaching a positively charged lipophilic substituent would enhance the antitumor activity of Met. In pursuit of this question, we synthesized a set of mitochondria-targeted Met analogues (Mito-Mets) with varying alkyl chain lengths containing a triphenylphosphonium cation (TPP+). In particular, the analogue Mito-Met10, synthesized by attaching TPP+ to Met via a 10-carbon aliphatic side chain, was nearly 1,000 times more efficacious than Met at inhibiting cell proliferation in pancreatic ductal adenocarcinoma (PDAC). Notably, in PDAC cells, Mito-Met10 potently inhibited mitochondrial complex I, stimulating superoxide and AMPK activation, but had no effect in nontransformed control cells. Moreover, Mito-Met10 potently triggered G1 cell-cycle phase arrest in PDAC cells, enhanced their radiosensitivity, and more potently abrogated PDAC growth in preclinical mouse models, compared with Met. Collectively, our findings show how improving the mitochondrial targeting of Met enhances its anticancer activities, including aggressive cancers like PDAC in great need of more effective therapeutic options. Cancer Res; 76(13); 3904–15. ©2016 AACR.
Antagonizing Integrin β3 Increases Immunosuppression in Cancer
Authors: Su Xinming , Esser Alison K. , Amend Sarah R. , Xiang Jingyu , Xu Yalin , Ross Michael H. , Fox Gregory C. , Kobayashi Takayuki , Steri Veronica , Roomp Kirsten , Fontana Francesca , Hurchla Michelle A. , Knolhoff Brett L. , Meyer Melissa A. , Morgan Elizabeth A. , Tomasson Julia C. , Novack Joshua S. , Zou Wei , Faccio Roberta , Novack Deborah V. , Robinson Stephen D. , Teitelbaum Steven L. , DeNardo David G. , Schneider Jochen G. , Weilbaecher Katherine N.
Published: May 2016

Integrin β3 is critical for tumor invasion, neoangiogenesis, and inflammation, making it a promising cancer target. However, preclinical and clinical data of integrin β3 antagonists have demonstrated no benefit or worse outcomes. We hypothesized that integrin β3 could affect tumor immunity and evaluated tumors in mice with deletion of integrin β3 in macrophage lineage cells (β3KOM). β3KOM mice had increased melanoma and breast cancer growth with increased tumor-promoting M2 macrophages and decreased CD8+ T cells. Integrin β3 antagonist, cilengitide, also enhanced tumor growth and increased M2 function. We uncovered a negative feedback loop in M2 myeloid cells, wherein integrin β3 signaling favored STAT1 activation, an M1-polarizing signal, and suppressed M2-polarizing STAT6 activation. Finally, disruption of CD8+ T cells, macrophages, or macrophage integrin β3 signaling blocked the tumor-promoting effects of integrin β3 antagonism. These results suggest that effects of integrin β3 therapies on immune cells should be considered to improve outcomes. Cancer Res; 76(12); 3484–95. ©2016 AACR.
The Tumor-Associated Glycosyltransferase ST6Gal-I Regulates Stem Cell Transcription Factors and Confers a Cancer Stem Cell Phenotype
Authors: Schultz Matthew J. , Holdbrooks Andrew T. , Chakraborty Asmi , Grizzle William E. , Landen Charles N. , Buchsbaum Donald J. , Conner Michael G. , Arend Rebecca C. , Yoon Karina J. , Klug Christopher A. , Bullard Daniel C. , Kesterson Robert A. , Oliver Patsy G. , O'Connor Amber K. , Yoder Bradley K. , Bellis Susan L.
Published: May 2016

The glycosyltransferase ST6Gal-I, which adds α2-6–linked sialic acids to substrate glycoproteins, has been implicated in carcinogenesis; however, the nature of its pathogenic role remains poorly understood. Here we show that ST6Gal-I is upregulated in ovarian and pancreatic carcinomas, enriched in metastatic tumors, and associated with reduced patient survival. Notably, ST6Gal-I upregulation in cancer cells conferred hallmark cancer stem-like cell (CSC) characteristics. Modulating ST6Gal-I expression in pancreatic and ovarian cancer cells directly altered CSC spheroid growth, and clonal variants with high ST6Gal-I activity preferentially survived in CSC culture. Primary ovarian cancer cells from patient ascites or solid tumors sorted for α2-6 sialylation grew as spheroids, while cells lacking α2-6 sialylation remained as single cells and lost viability. ST6Gal-I also promoted resistance to gemcitabine and enabled the formation of stably resistant colonies. Gemcitabine treatment of patient-derived xenograft tumors enriched for ST6Gal-I–expressing cells relative to pair-matched untreated tumors. ST6Gal-I also augmented tumor-initiating potential. In limiting dilution assays, subcutaneous tumor formation was inhibited by ST6Gal-I knockdown, whereas in a chemically induced tumor initiation model, mice with conditional ST6Gal-I overexpression exhibited enhanced tumorigenesis. Finally, we found that ST6Gal-I induced expression of the key tumor-promoting transcription factors, Sox9 and Slug. Collectively, this work highlighted a previously unrecognized role for a specific glycosyltransferase in driving a CSC state. Cancer Res; 76(13); 3978–88. ©2016 AACR.
Rho GTPase Transcriptome Analysis Reveals Oncogenic Roles for Rho GTPase-Activating Proteins in Basal-like Breast Cancers
Authors: Lawson Campbell D. , Fan Cheng , Mitin Natalia , Baker Nicole M. , George Samuel D. , Graham David M. , Perou Charles M. , Burridge Keith , Der Channing J. , Rossman Kent L.
Published: May 2016

The basal-like breast cancer (BLBC) subtype accounts for a disproportionately high percentage of overall breast cancer mortality. The current therapeutic options for BLBC need improvement; hence, elucidating signaling pathways that drive BLBC growth may identify novel targets for the development of effective therapies. Rho GTPases have previously been implicated in promoting tumor cell proliferation and metastasis. These proteins are inactivated by Rho-selective GTPase-activating proteins (RhoGAP), which have generally been presumed to act as tumor suppressors. Surprisingly, RNA-Seq analysis of the Rho GTPase signaling transcriptome revealed high expression of several RhoGAP genes in BLBC tumors, raising the possibility that these genes may be oncogenic. To evaluate this, we examined the roles of two of these RhoGAPs, ArhGAP11A (also known as MP-GAP) and RacGAP1 (also known as MgcRacGAP), in promoting BLBC. Both proteins were highly expressed in human BLBC cell lines, and knockdown of either gene resulted in significant defects in the proliferation of these cells. Knockdown of ArhGAP11A caused CDKN1B/p27-mediated arrest in the G1 phase of the cell cycle, whereas depletion of RacGAP1 inhibited growth through the combined effects of cytokinesis failure, CDKN1A/p21-mediated RB1 inhibition, and the onset of senescence. Random migration was suppressed or enhanced by the knockdown of ArhGAP11A or RacGAP1, respectively. Cell spreading and levels of GTP-bound RhoA were increased upon depletion of either RhoGAP. We have established that, via the suppression of RhoA, ArhGAP11A and RacGAP1 are both critical drivers of BLBC growth, and propose that RhoGAPs can act as oncogenes in cancer. Cancer Res; 76(13); 3826–37. ©2016 AACR.
FAP Promotes Immunosuppression by Cancer-Associated Fibroblasts in the Tumor Microenvironment via STAT3–CCL2 Signaling
Authors: Yang Xuguang , Lin Yuli , Shi Yinghong , Li Bingji , Liu Weiren , Yin Wei , Dang Yongjun , Chu Yiwei , Fan Jia , He Rui
Published: May 2016

Cancer-associated fibroblasts (CAF) are components of the tumor microenvironment whose contributions to malignant progression are not fully understood. Here, we show that the fibroblast activation protein (FAP) triggers induction of a CAF subset with an inflammatory phenotype directed by STAT3 activation and inflammation-associated expression signature marked by CCL2 upregulation. Enforcing FAP expression in normal fibroblasts was sufficient to endow them with an inflammatory phenotype similar to FAP+CAFs. We identified FAP as a persistent activator of fibroblastic STAT3 through a uPAR-dependent FAK–Src–JAK2 signaling pathway. In a murine liver tumor model, we found that FAP+CAFs were a major source of CCL2 and that fibroblastic STAT3–CCL2 signaling in this setting promoted tumor growth by enhancing recruitment of myeloid-derived suppressor cells (MDSC). The CCL2 receptor CCR2 was expressed on circulating MDSCs in tumor-bearing subjects and FAP+CAF-mediated tumor promotion and MDSC recruitment was abrogated in Ccr2-deficient mice. Clinically, we observed a positive correlation between stromal expression of FAP, p-STAT3, and CCL2 in human intrahepatic cholangiocarcinoma, a highly aggressive liver cancer with dense desmoplastic stroma, where elevated levels of stromal FAP predicted a poor survival outcome. Taken together, our results showed how FAP–STAT3–CCL2 signaling in CAFs was sufficient to program an inflammatory component of the tumor microenvironment, which may have particular significance in desmoplasia-associated cancers. Cancer Res; 76(14); 4124–35. ©2016 AACR.
Clonotypic Diversification of Intratumoral T Cells Following Sipuleucel-T Treatment in Prostate Cancer Subjects
Authors: Sheikh Nadeem , Cham Jason , Zhang Li , DeVries Todd , Letarte Simon , Pufnock Jeff , Hamm David , Trager James , Fong Lawrence
Published: May 2016

Sipuleucel-T is an autologous cellular therapy for asymptomatic, or minimally symptomatic, metastatic castrate-resistant prostate cancer, designed to stimulate an immune response against prostate cancer. In a recent clinical trial (NCT00715104), we found that neoadjuvant sipuleucel-T increased the number of activated T cells within the tumor microenvironment. The current analysis examined whether sipuleucel-T altered adaptive T-cell responses by expanding pre-existing T cells or by recruiting new T cells to prostate tissue. Next-generation sequencing of the T-cell receptor (TCR) genes from blood or prostate tissue was used to quantitate and track T-cell clonotypes in these treated subjects with prostate cancer. At baseline, there was a significantly greater diversity of circulating TCR sequences in subjects with prostate cancer compared with healthy donors. Among healthy donors, circulating TCR sequence diversity remained unchanged over the same time interval. In contrast, sipuleucel-T treatment reduced circulating TCR sequence diversity versus baseline as measured by the Shannon index. Interestingly, sipuleucel-T treatment resulted in greater TCR sequence diversity in resected prostate tissue in sipuleucel-T–treated subjects versus tissue of nonsipuleucel-T–treated subjects with prostate cancer. Furthermore, sipuleucel-T increased TCR sequence commonality between blood and resected prostate tissue in treated versus untreated subjects with prostate cancer. The broadening of the TCR repertoire within the prostate tissue supports the hypothesis that sipuleucel-T treatment facilitates the recruitment of T cells into the prostate. Our results highlight the importance of assessing T-cell response to immunotherapy both in the periphery and in tumor tissue. Cancer Res; 76(13); 3711–8. ©2016 AACR.
Adipocyte Exosomes Promote Melanoma Aggressiveness through Fatty Acid Oxidation: A Novel Mechanism Linking Obesity and Cancer
Authors: Lazar Ikrame , Clement Emily , Dauvillier Stéphanie , Milhas Delphine , Ducoux-Petit Manuelle , LeGonidec Sophie , Moro Cédric , Soldan Vanessa , Dalle Stéphane , Balor Stéphanie , Golzio Muriel , Burlet-Schiltz Odile , Valet Philippe , Muller Catherine , Nieto Laurence
Published: May 2016

Malignant progression results from a dynamic cross-talk between stromal and cancer cells. Recent evidence suggests that this cross-talk is mediated to a significant extent by exosomes, nanovesicles secreted by most cell types and which allow the transfer of proteins, lipids, and nucleic acids between cells. Adipocytes are a major component of several tumor microenvironments, including that of invasive melanoma, where cells have migrated to the adipocyte-rich hypodermic layer of the skin. We show that adipocytes secrete exosomes in abundance, which are then taken up by tumor cells, leading to increased migration and invasion. Using mass spectrometry, we analyzed the proteome of adipocyte exosomes. Interestingly, these vesicles carry proteins implicated in fatty acid oxidation (FAO), a feature highly specific to adipocyte exosomes. We further show that, in the presence of adipocyte exosomes, FAO is increased in melanoma cells. Inhibition of this metabolic pathway completely abrogates the exosome-mediated increase in migration. Moreover, in obese mice and humans, both the number of exosomes secreted by adipocytes as well as their effect on FAO-dependent cell migration are amplified. These observations might in part explain why obese melanoma patients have a poorer prognosis than their nonobese counterparts. Cancer Res; 76(14); 4051–7. ©2016 AACR.
Anti-EGFR Targeted Monoclonal Antibody Isotype Influences Antitumor Cellular Immunity in Head and Neck Cancer Patients
Authors: Trivedi Sumita , Srivastava Raghvendra M. , Concha-Benavente Fernando , Ferrone Soldano , Garcia-Bates Tatiana M. , Li Jing , Ferris Robert L.
Published: May 2016

Purpose: EGF receptor (EGFR) is highly overexpressed on several cancers and two targeted anti-EGFR antibodies which differ by isotype are FDA-approved for clinical use. Cetuximab (IgG1 isotype) inhibits downstream signaling of EGFR and activates antitumor, cellular immune mechanisms. As panitumumab (IgG2 isotype) may inhibit downstream EGFR signaling similar to cetuximab, it might also induce adaptive immunity.

Experimental Design: We measured in vitro activation of cellular components of the innate and adaptive immune systems. We also studied the in vivo activation of components of the adaptive immune system in patient specimens from two recent clinical trials using cetuximab or panitumumab.

Results: Both monoclonal antibodies (mAb) primarily activate natural killer (NK) cells, although cetuximab is significantly more potent than panitumumab. Cetuximab-activated neutrophils mediate antibody-dependent cellular cytotoxicity (ADCC) against head and neck squamous cell carcinomas (HNSCC) tumor cells, and interestingly, this effect was FcγRIIa- and FcγRIIIa genotype–dependent. Panitumumab may activate monocytes through CD32 (FcγRIIa); however, monocytes activated by either mAb are not able to mediate ADCC. Cetuximab enhanced dendritic cell (DC) maturation to a greater extent than panitumumab, which was associated with improved tumor antigen cross-presentation by cetuximab compared with panitumumab. This correlated with increased EGFR-specific cytotoxic CD8+ T cells in patients treated with cetuximab compared with those treated with panitumumab.

Conclusions: Although panitumumab effectively inhibits EGFR signaling to a similar extent as cetuximab, it is less effective at triggering antitumor, cellular immune mechanisms which may be crucial for effective therapy of HNSCC. Clin Cancer Res; 22(21); 5229–37. ©2016 AACR.
Misoplegia
Authors: Pearce J.M.S.
Published: May 2016

Amongst the many fascinating abnormal perceptions of parts of the body which may follow strokes and other cerebral lesions is the rare phenomenon of misoplegia. Afflicted patients show dislike, amounting to hatred of the affected part, often accompanied by verbal or physical abuse. Most often the result of a right hemisphere lesion, its precise mechanisms are not well understood. This is a summary of misoplegia and its alleged physiological mechanisms.
Two Common Reasons of Malabsorption Syndromes: Celiac Disease and Whipple’s Disease
Authors: Sun Dan-Feng , Fang Jing-Yuan
Published: May 2016

Malabsorption syndromes are commonly caused by pathological interferences of normal digestive processes. In the last several years major advances in the diagnosis and treatment of these syndromes has emerged. This review will focus on diseases in which the mucosal phase of the digestive process is predominately disturbed, including celiac disease and Whipple’s disease. Since most diagnostic tests have a limited availability, it will also provide a diagnostic algorithm of malabsorption syndromes.
Diagnostic Value of the Reflux Disease Questionnaire in General Practice
Authors: Aanen M.C. , Numans M.E. , Weusten B.L.A.M. , Smout A.J.P.M.
Published: May 2016

Objectives: This study determined the diagnostic and therapeutic response of the Reflux Disease Questionnaire (RDQ) using the symptom association probability (SAP) as reference. In addition, the RDQ’s construct validity and its relationship to quality of life (QOL) were ascertained. Methods: Seventy-four patients with GORD symptoms (age 51 years (22–78); ♂62%) derived from primary care completed the RDQ, GSRS and QOLRAD before and after a 2 weeks’ course of esomeprazole 40 mg daily. The SAP was determined by a 24-hour pH recording before PPI treatment. The diagnostic abilities of the RDQ (total and 4 dimensions scores) were assessed with the area under the curve (AUC) of a receiver operating curve. RDQ scores before and after PPI treatment were compared with Wilcoxon tests. Multiple linear regressions assessed the RDQ’s construct validity (GSRS) and relationship to QOL (QOLRAD). Results: The AUCs were low for all RDQ dimensions (AUC <0.6). In the SAP-positive patients all RDQ dimensions improved (p < 0.0001) while the scores of the SAP negatives did not (heartburn p < 0.01; GORD and total score p < 0.05; regurgitation and dyspepsia n.s.). The RDQ was related to the total and reflux GSRS dimensions while the food and drink QOL dimension was linearly associated with the RDQ. Conclusions: The RDQ is a valid and reliable questionnaire with excellent construct validity and a good relationship to QOL. The diagnostic value of the RDQ in primary care is limited, but combination with an additional PPI treatment course might improve the RDQ’s ability to discriminate GORD patients according to their SAP outcome.
Cytotoxicity of Diospyrin and Its Derivatives in Relation to the Generation of Reactive Oxygen Species in Tumour Cells in vitroandin vivo
Authors: Hazra B. , Das Sarma M. , Kumar B. , Basu S. , Das K. , Pandey B.N. , Mishra K.P.
Published: May 2016

Background: Alkyl ethers (D2 and D7) synthesized from diospyrin (D1), a naphthoquinonoid isolated from Diospyros montana Roxb., were evaluated for cytotoxicity and capacity to generate reactive oxygen species (ROS) in tumour cells. Methods: The tumour inhibitory activity of the quinonoids was assessed in vivo against Ehrlich ascites carcinoma (EAC), while cytotoxicity was determined in vitro on EAC and MCF-7 cancer cells by MTT assay. ROS generated by quinonoids in MCF-7 cells was measured fluorimetrically. Results: The tumour inhibitory activity, cytotoxicity and ROS generation capacity of quinonoids were found to be D7 > D2 > D1. Conclusion:Alkyl ethers of D1 were more cytotoxic against tumour cells in vitro as well as in vivo.
Vivotif® – A ‘Magic Shield’ for Protection against Typhoid Fever and Delivery of Heterologous Antigens
Authors: Gentschev Ivaylo , Spreng Simone , Sieber Heike , Ures Jose , Mollet Fabian , Collioud Andre , Pearman Jon , Griot-Wenk Monika E. , Fensterle Joachim , Rapp Ulf R. , Goebel Werner , Rothen Simon A. , Dietrich Guido
Published: May 2016

The attenuated Salmonella typhi strain Ty21a is the main constituent of Vivotif®, the only attenuated live oral vaccine against typhoid fever. In comparison with antibiotics, the ‘magic bullets’ which Paul Ehrlich was striving for to treat infectious diseases, this vaccine should be viewed as a ‘magic shield’, because rather than treating typhoid fever after the infection has started, immunisation with this vaccine strain prevents infection and disease by the induction of specific immune responses. Ty21a is also an attractive carrier for the delivery of heterologous antigens. Recently, we successfully used Ty21a for antigen delivery via the haemolysin secretion system of Escherichia coli, which allows efficient protein secretion from the carrier bacteria.