bluuebunny/crossref_metadata_2025_split huggingface dataset:
Determinants of visual acuity outcomes in eyes with neovascular AMD treated with anti-VEGF agents: an instrumental variable analysis of the AURA study
Authors: Holz F G , Tadayoni R , Beatty S , Berger A R , Cereda M G , Hykin P , Staurenghi G , Wittrup-Jensen K , Nilsson J , Kim K , Sivaprasad S 
Published: May 2016

Abstract

Purpose
To identify the strongest variable(s) linked with the number of ranibizumab injections and outcomes in AURA, and to identify ways to improve outcomes using this association.

Methods
AURA was a large observational study that monitored visual acuity over a 2-year period in patients with neovascular age-related macular degeneration (AMD) who received ranibizumab injections. Baseline characteristics, resource use, and outcomes were analyzed using an instrumental variable approach and regression analysis.

Results
Data were analyzed from 2227 patients enrolled in AURA. Optical coherence tomography (OCT) and ophthalmoscopy were the most common diagnostic tests used, and this combination was the strongest instrumental variable. Use of OCT and ophthalmoscopy affected the number of injections given and resulted in an increase in visual acuity gains from baseline of 17.6 letters in year 1 and 2.5 letters in year 2. Regression models using the instrumental variable (OCT and ophthalmoscopy combined) showed that ≥5.1 (95% CI: 3.3–11.4) ranibizumab injections were needed to maintain visual acuity from baseline to year 1 and ≥8.3 (95% CI: 5.3–18.8) injections were needed to maintain visual acuity from year 1 to year 2. To gain ≥15 letters, ≥7.9 (95% CI: 5.1–17.5) ranibizumab injections would be needed in year 1 and ≥16.1 (95% CI: 10.3–36.4) injections would be needed over 2 years.

Conclusions
These findings highlight the role that regular monitoring plays in guiding neovascular AMD therapy and they showed that the number of ranibizumab injections needed to maintain visual acuity is higher than that administered in AURA.
Myoscape controls cardiac calcium cycling and contractility via regulation of L-type calcium channel surface expression
Authors: Eden Matthias , Meder Benjamin , Völkers Mirko , Poomvanicha Montatip , Domes Katrin , Branchereau M. , Marck P. , Will Rainer , Bernt Alexander , Rangrez Ashraf , Busch Matthias ,   German Mouse Clinic Consortium, Adler Thure , Busch Dirk H. , Antonio Aguilar-Pimentel Juan , Ollert Markus , Götz Alexander , Schulz Holger , Prehn Cornelia , Adamski Jerzy , Becker Lore , Klopstock Thomas , Horsch Marion , Beckers Johannes , Schrewe Anja , Bekeredjian Raffi , Katus Hugo , Garrett Lillian , Hölter Sabine M. , Wurst Wolfgang , Puk Oliver , Graw Jochen , Hans Wolfgang , Rozman Jan , Klingenspor Martin , Neff Frauke , Tost Monica , Calzada-Wack Julia , Klein-Rodewald Tanja , Rácz Ildikó , Zimmer Andreas , Rathkolb Birgit , Wolf Eckhard , Lengger Christoph , Maier Holger , Stoeger Claudia , Leuchtenberger Stefanie , Gailus-Durner Valéri , Fuchs Helmut , Hrabě de Angelis Martin , Heymes Christophe , Rottbauer Wolfgang , Most Patrick , Hofmann Franz , Frey Norbert 
Published: May 2016

AbstractCalcium signalling plays a critical role in the pathogenesis of heart failure. Here we describe a cardiac protein named Myoscape/FAM40B/STRIP2, which directly interacts with the L-type calcium channel. Knockdown of Myoscape in cardiomyocytes decreases calcium transients associated with smaller Ca2+ amplitudes and a lower diastolic Ca2+ content. Likewise, L-type calcium channel currents are significantly diminished on Myoscape ablation, and downregulation of Myoscape significantly reduces contractility of cardiomyocytes. Conversely, overexpression of Myoscape increases global Ca2+ transients and enhances L-type Ca2+ channel currents, and is sufficient to restore decreased currents in failing cardiomyocytes. In vivo, both Myoscape-depleted morphant zebrafish and Myoscape knockout (KO) mice display impairment of cardiac function progressing to advanced heart failure. Mechanistically, Myoscape-deficient mice show reduced L-type Ca2+currents, cell capacity and calcium current densities as a result of diminished LTCC surface expression. Finally, Myoscape expression is reduced in hearts from patients suffering of terminal heart failure, implying a role in human disease.
Live single-cell laser tag
Authors: Binan Loïc , Mazzaferri Javier , Choquet Karine , Lorenzo Louis-Etienne , Wang Yu Chang , Affar El Bachir , De Koninck Yves , Ragoussis Jiannis , Kleinman Claudia L. , Costantino Santiago 
Published: May 2016

AbstractThe ability to conduct image-based, non-invasive cell tagging, independent of genetic engineering, is key to cell biology applications. Here we introduce cell labelling via photobleaching (CLaP), a method that enables instant, specific tagging of individual cells based on a wide array of criteria such as shape, behaviour or positional information. CLaP uses laser illumination to crosslink biotin onto the plasma membrane, coupled with streptavidin conjugates to label individual cells for genomic, cell-tracking, flow cytometry or ultra-microscopy applications. We show that the incorporated mark is stable, non-toxic, retained for several days, and transferred by cell division but not to adjacent cells in culture. To demonstrate the potential of CLaP for genomic applications, we combine CLaP with microfluidics-based single-cell capture followed by transcriptome-wide next-generation sequencing. Finally, we show that CLaP can also be exploited for inducing transient cell adhesion to substrates for microengineering cultures with spatially patterned cell types.
Gut environment-induced intraepithelial autoreactive CD4+ T cells suppress central nervous system autoimmunity via LAG-3
Authors: Kadowaki Atsushi , Miyake Sachiko , Saga Ryoko , Chiba Asako , Mochizuki Hideki , Yamamura Takashi 
Published: May 2016

AbstractThe gut environment has been found to significantly influence autoimmune diseases such as multiple sclerosis; however, immune cell mechanisms are unclear. Here we show that the gut epithelium of myelin oligodendrocyte glycoprotein(35-55)-specific T-cell receptor transgenic mice contains environmental stimuli-induced intraepithelial lymphocytes (IELs) that inhibit experimental autoimmune encephalomyelitis on transfer. These cells express surface markers phenotypical of ‘induced’ IELs, have a TH17-like profile and infiltrate the central nervous system (CNS). They constitutively express Ctla4 and Tgfb1 and markedly upregulate Lag3 expression in the CNS, thereby inhibiting inflammation. We also demonstrate the suppressive capability of CD4+ IELs with alternative antigen specificities, their proliferation in response to gut-derived antigens and contribution of the microbiota and dietary aryl hydrocarbon receptor ligands to their induction. Thus, the gut environment favours the generation of autoreactive CD4+ T cells with unique regulatory functions, potentially important for preventing CNS autoimmunity.
Graphene Oxide promotes embryonic stem cell differentiation to haematopoietic lineage
Authors: Garcia-Alegria Eva , Iliut Maria , Stefanska Monika , Silva Claudio , Heeg Sebastian , Kimber Susan J. , Kouskoff Valerie , Lacaud Georges , Vijayaraghavan Aravind , Batta Kiran 
Published: May 2016

AbstractPluripotent stem cells represent a promising source of differentiated tissue-specific stem and multipotent progenitor cells for regenerative medicine and drug testing. The realisation of this potential relies on the establishment of robust and reproducible protocols of differentiation. Several reports have highlighted the importance of biomaterials in assisting directed differentiation. Graphene oxide (GO) is a novel material that has attracted increasing interest in the field of biomedicine. In this study, we demonstrate that GO coated substrates significantly enhance the differentiation of mouse embryonic stem (ES) cells to both primitive and definitive haematopoietic cells. GO does not affect cell proliferation or survival of differentiated cells but rather enhances the transition of haemangioblasts to haemogenic endothelial cells, a key step during haematopoietic specification. Importantly, GO also improves, in addition to murine, human ES cell differentiation to blood cells. Taken together, our study reveals a positive role for GO in haematopoietic differentiation and suggests that further functionalization of GO could represent a valid strategy for the generation of large numbers of functional blood cells. Producing these cells would accelerate haematopoietic drug toxicity testing and treatment of patients with blood disorders or malignancies.
Monitoring live human mesenchymal stromal cell differentiation and subsequent selection using fluorescent RNA-based probes
Authors: Li Bojun , Menzel Ursula , Loebel Claudia , Schmal Hagen , Alini Mauro , Stoddart Martin J. 
Published: May 2016

AbstractInvestigating mesenchymal stromal cell differentiation requires time and multiple samples due to destructive endpoint assays. Osteogenesis of human bone marrow derived mesenchymal stromal cells (hBMSCs) has been widely studied for bone tissue engineering. Recent studies show that the osteogenic differentiation of hBMSCs can be assessed by quantifying the ratio of two important transcription factors (Runx2/Sox9). We demonstrate a method to observe mRNA expression of two genes in individual live cells using fluorescent probes specific for Runx2 and Sox9 mRNA. The changes of mRNA expression in cells can be observed in a non-destructive manner. In addition, the osteogenic hBMSCs can be prospectively identified and obtained based on the relative intracellular fluorescence of Sox9 in relation to Runx2 using fluorescence activated cell sorting. Relatively homogeneous cell populations with high osteogenic potential can be isolated from the original heterogeneous osteogenically induced hBMSCs within the first week of induction. This offers a more detailed analysis of the effectiveness of new therapeutics both at the individual cell level and the response of the population as a whole. By identifying and isolating differentiating cells at early time points, prospective analysis of differentiation is also possible, which will lead to a greater understanding of MSC differentiation.
Functional assessment of glioma pathogenesis by in vivo multi-parametric magnetic resonance imaging and in vitro analyses
Authors: Yao Nai-Wei , Chang Chen , Lin Hsiu-Ting , Yen Chen-Tung , Chen Jeou-Yuan 
Published: May 2016

AbstractGliomas are aggressive brain tumors with poor prognosis. In this study, we report a novel approach combining bothin vivomulti-parametric MRI andin vitrocell culture assessments to evaluate the pathogenic development of gliomas. Osteopontin (OPN), a pleiotropic factor, has been implicated in the formation and progression of various human cancers, including gliomas, through its functions in regulating cell proliferation, survival, angiogenesis and migration. Using rat C6 glioma model, the combined approach successfully monitors the acquisition and decrease of cancer hallmarks. We show that knockdown of the expression ofOPNreduces C6 cell proliferation, survival, viability and clonogenicityin vitroand reduces tumor burden and prolongs animal survival in syngeneic rats.OPNdepletion is associated with reduced tumor growth, decreased angiogenesis and an increase of tumor-associated metabolites, as revealed by T2-weighted images, diffusion-weighted images, Ktransmaps and 1H-MRS, respectively. These strategies allow us to define an important role of OPN in conferring cancer hallmarks, which can be further applied to assess the functional roles of other candidate genes in glioma. In particular, the non-invasive multi-parametric MRI measurement of cancer hallmarks related to proliferation, angiogenesis and altered metabolism may serve as a useful tool for diagnosis and for patient management.
The DEAD-box RNA helicase 51 controls non-small cell lung cancer proliferation by regulating cell cycle progression via multiple pathways
Authors: Wang Xiaojing , Liu Hongli , Zhao Chengling , Li Wei , Xu Huanbai , Chen Yuqing 
Published: May 2016

AbstractThe genetic regulation of cell cycle progression and cell proliferation plays a role in the growth of non-small cell lung cancer (NSCLC), one of the most common causes of cancer-related mortality. Although DEAD-box RNA helicases are known to play a role in cancer development, including lung cancer, the potential involvement of the novel family member DDX51 has not yet been investigated. In the current study we assessed the role of DDX51 in NSCLC using a siRNA-based approach. DDX51 siRNA-expressing cells exhibited a slower cell proliferation rate and underwent arrest in S-phase of the cell cycle compared with control cells. Microarray analyses revealed that DDX51siRNA expression resulted in the dysregulation of a number of cell signalling pathways. Moreover, injection of DDX51 siRNA into an animal model resulted in the formation of smaller tumours compared with the control group. We also assessed the expression of DDX51 in patients with NSCLC, and the data revealed that the expression was correlated with patient age but no other risk factors. Overall, our data suggest for the first time that DDX51 aids cell cancer proliferation by regulating multiple signalling pathways, and that this protein might be a therapeutic target for NSCLC.
Distinctive proteomic profiles among different regions of human carotid plaques in men and women
Authors: Liang Wenzhao , Ward Liam J. , Karlsson Helen , Ljunggren Stefan A. , Li Wei , Lindahl Mats , Yuan Xi-Ming 
Published: May 2016

AbstractThe heterogeneity of atherosclerotic tissue has limited comprehension in proteomic and metabolomic analyses. To elucidate the functional implications and differences between genders, of atherosclerotic lesion formation we investigated protein profiles from different regions of human carotid atherosclerotic arteries; internal control, fatty streak, plaque shoulder, plaque centre and fibrous cap. Proteomic analysis was performed using 2-DE with MALDI-TOF, with validation using nLC-MS/MS. Protein mapping of 2-DE identified 52 unique proteins, including 15 previously unmapped proteins, of which 41 proteins were confirmed by nLC-MS/MS analysis. Expression levels of 18 proteins were significantly altered in plaque regions compared to the internal control region. Nine proteins showed site-specific alterations, irrespective of gender, with clear associations to extracellular matrix remodelling. Five proteins display gender-specific alterations with 2-DE, with two alterations validated by nLC-MS/MS. Gender differences in ferritin light chain and transthyretin were validated using both techniques. Validation of immunohistochemistry confirmed significantly higher levels of ferritin in plaques from male patients. Proteomic analysis of different plaque regions has reduced the effects of plaque heterogeneity and significant differences in protein expression are determined in specific regions and between genders. These proteomes have functional implications in plaque progression and are of importance in understanding gender differences in atherosclerosis.
A bulk sub-femtoliter in vitro compartmentalization system using super-fine electrosprays
Authors: Sharma Bineet , Takamura Yuzuru , Shimoda Tatsuya , Biyani Manish 
Published: May 2016

AbstractThe extreme miniaturization of biological and chemical assays in aqueous-droplet compartments enables spatiotemporal control for large-scale parallel experimentation and can thus permit new capabilities for “digitizing” directed molecular evolution methodologies. We report a remarkably facile bulk method to generate mega-scale monodisperse sub-femtoliter aqueous droplets by electrospray, using a prototype head with super-fine inkjet technology. Moreover, the electrostatic inkjet nozzle that injects the aqueous phase when immersed within an immiscible phase (an optimized oil/surfactant mixture) has the advantage of generating cell-like sub-femtoliter compartments for biomolecule encapsulation and successive biological and chemical reactions. Sub-femtoliter droplets of both liquid (water-in-oil, volumes ranging from 0.2 to 6.4 fL) and gel bead (agarose-in-oil, volume ranging from 0.3 to 15.6 fL) compartments with average sizes of 1.3 μm and 1.5 μm, respectively, were successfully generated using an inkjet nozzle at a speed of more than 105 droplets per second. We demonstrated the applicability of this system by synthesizing fluorescent proteins using a cell-free expression system inside electrosprayed sub-femtoliter droplets at an accelerated rate, thereby extending the utility of in vitro compartmentalization with improved analytical performance for a top-down artificial cellular system.
Fly DPP10 acts as a channel ancillary subunit and possesses peptidase activity
Authors: Shiina Yohei , Muto Tomohiro , Zhang Zhili , Baihaqie Ahmad , Yoshizawa Takamasa , Lee Hye-in J. , Park Eulsoon , Tsukiji Shinya , Takimoto Koichi 
Published: May 2016

AbstractMammalian DPP6 (DPPX) and DPP10 (DPPY) belong to a family of dipeptidyl peptidases, but lack enzyme activity. Instead, these proteins form complexes with voltage-gated K+ channels in Kv4 family to control their gating and other properties. Here, we find that the fly DPP10 ortholog acts as an ancillary subunit of Kv4 channels and digests peptides. Similarly to mammalian DPP10, the fly ortholog tightly binds to rat Kv4.3 protein. The association causes negative shifts in voltage dependence of channel activation and steady state inactivation. It also results in faster inactivation and recovery from inactivation. In addition to its channel regulatory role, fly DPP10 exhibits significant dipeptidyl peptidase activity with Gly-Pro-MCA (glycyl-L-proline 4-methylcoumaryl-7-amide) as a substrate. Heterologously expressed Flag-tagged fly DPP10 and human DPP4 show similar Km values towards this substrate. However, fly DPP10 exhibits approximately a 6-times-lower relative kcat value normalized with anti-Flag immunoreactivity than human DPP4. These results demonstrate that fly DPP10 is a dual functional protein, controlling Kv4 channel gating and removing bioactive peptides.
RIG-like Helicase Regulation of Chitinase 3-like 1 Axis and Pulmonary Metastasis
Authors: Ma Bing , Herzog Erica L. , Moore Meagan , Lee Chang-Min , Na Sung Hun , Lee Chun Geun , Elias Jack A. 
Published: May 2016

AbstractChi3l1 is induced by a variety of cancers where it portends a poor prognosis and plays a key role in the generation of metastasis. However, the mechanisms that Chi3l1 uses to mediate these responses and the pathways that control Chi3l1-induced tumor responses are poorly understood. We characterized the mechanisms that Chi3l1 uses to foster tumor progression and the ability of the RIG-like helicase (RLH) innate immune response to control Chi3l1 elaboration and pulmonary metastasis. Here we demonstrate that RLH activation inhibits tumor induction of Chi3l1 and the expression of receptor IL-13Rα2 and pulmonary metastasis while restoring NK cell accumulation and activation, augmenting the expression of IFN-α/β, chemerin and its receptor ChemR23, p-cofilin, LIMK2 and PTEN and inhibiting BRAF and NLRX1 in a MAVS-dependent manner. These studies demonstrate that Chi3l1 is a multifaceted immune stimulator of tumor progression and metastasis whose elaboration and tissue effects are abrogated by RLH innate immune responses.
Predicting global invasion risks: a management tool to prevent future introductions
Authors: Fletcher D. H. , Gillingham P. K. , Britton J. R. , Blanchet S. , Gozlan R. E. 
Published: May 2016

AbstractPredicting regions at risk from introductions of non-native species and the subsequent invasions is a fundamental aspect of horizon scanning activities that enable the development of more effective preventative actions and planning of management measures. The Asian cyprinid fish topmouth gudgeonPseudorasbora parvahas proved highly invasive across Europe since its introduction in the 1960s. In addition to direct negative impacts on native fish populations,P. parvahas potential for further damage through transmission of an emergent infectious disease, known to cause mortality in other species. To quantify its invasion risk, in regions where it has yet to be introduced, we trained 900 ecological niche models and constructed an Ensemble Model predicting suitability, then integrated a proxy for introduction likelihood. This revealed high potential forP. parvato invade regions well beyond its current invasive range. These included areas in all modelled continents, with several hotspots of climatic suitability and risk of introduction. We believe that these methods are easily adapted for a variety of other invasive species and that such risk maps could be used by policy-makers and managers in hotspots to formulate increased surveillance and early-warning systems that aim to prevent introductions and subsequent invasions.
Imaging Tumor Vascularity and Response to Anti-Angiogenic Therapy Using Gaussia Luciferase
Authors: Kantar Rami S. , Lashgari Ghazal , Tabet Elie I. , Lewandrowski Grant K. , Carvalho Litia A. , Tannous Bakhos A. 
Published: May 2016

AbstractWe developed a novel approach to assess tumor vascularity using recombinant Gaussia luciferase (rGluc) protein and bioluminescence imaging. Upon intravenous injection of rGluc followed by its substrate coelenterazine, non-invasive visualization of tumor vascularity by bioluminescence imaging was possible. We applied this method for longitudinal monitoring of tumor vascularity in response to the anti-angiogenic drug tivozanib. This simple and sensitive method could be extended to image blood vessels/vasculature in many different fields.
Points, skyrmions and torons in chiral nematic droplets
Authors: Posnjak Gregor , Čopar Simon , Muševič Igor 
Published: May 2016

AbstractChiral nematic droplets with perpendicular surface alignment of liquid crystalline molecules frustrate the helical structure into convoluted 3D textures with complex topology. We observe the droplets with fluorescent confocal polarising microscopy (FCPM) and reconstruct and analyse for the first time the topology of the 3D director field using a novel method of director reconstruction from raw data. We always find an odd number of topological defects, which preserve the total topological charge of the droplet of +1 regardless of chirality. At higher chirality, we observe up to 5 point hedgehog defects, which are elastically stabilized with convoluted twisted structures, reminiscent of 2D skyrmions and toron-like structure, nested into a sphere.
Embryonic development and inviability phenotype of chicken-Japanese quail F1 hybrids
Authors: Ishishita Satoshi , Kinoshita Keiji , Nakano Mikiharu , Matsuda Yoichi 
Published: May 2016

AbstractInterspecific hybrid incompatibility, including inviability and sterility, is important in speciation; however, its genetic basis remains largely unknown in vertebrates. Crosses between male chickens and female Japanese quails using artificial insemination can generate intergeneric hybrids; however, the hatching rate is low and hatched hybrids are only sterile males. Hybrid development is arrested frequently during the early embryonic stages and the sex ratio of living embryos is male-biased. However, the development and sex ratio of hybrid embryos have not been comprehensively analyzed. In the present study, we observed delayed embryonic development of chicken-quail hybrids during the early stage, compared with that of chickens and quails. The survival rate of hybrids decreased markedly during the blastoderm-to-pre-circulation stage and then decreased gradually through the subsequent stages. Hybrid females were observed at more than 10 d of incubation; however, the sex ratio of hybrids became male-biased from 10 d of incubation. Severely malformed embryos were observed frequently in hybrids. These results suggest that developmental arrest occurs at various stages in hybrid embryos, including a sexually non-biased arrest during the early stage and a female-biased arrest during the late stage. We discuss the genetic basis for hybrid inviability and its sex bias.