bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluation of Eluxadoline Effect on Cardiac Repolarization
Authors: Bonifacio Laura , Hunt Thomas L. , McIntyre Gail , Dove Leonard S. , Covington Paul S. 
Published: April 2018

AbstractThis study evaluated the effects of eluxadoline, a mixed μ‐opioid receptor (OR) and κ‐OR agonist and δ‐OR antagonist, on cardiac repolarization. This evaluator‐blinded, placebo‐ and positive‐controlled, 4‐period crossover study randomized healthy men and women to single oral doses of eluxadoline (therapeutic dose 100 mg or supratherapeutic dose 1000 mg), moxifloxacin 400 mg, or placebo. QT data were corrected using individual custom correction (QTcI). The primary endpoint was the change from baseline in QTcI intervals (ΔQTcI) between eluxadoline and placebo (ΔΔQTcI). An upper bound of the 95% confidence interval around ΔΔQTcI of 10 milliseconds was considered clinically significant. Concentration–QTc data were analyzed using a repeated‐measures, mixed‐effects linear model. Sixty‐four volunteers were treated, and 58 completed the study. Assay sensitivity was demonstrated with moxifloxacin (noted by ΔΔQTcI of 11.94 milliseconds). The maximum ΔΔQTcI for eluxadoline 1000 mg was 4.10 milliseconds 1 hour postdose (1‐sided 95% upper confidence bound, 5.81 milliseconds), and for eluxadoline 100 mg was 1.20 milliseconds at 0.5 hours postdose (1‐sided 95% upper confidence bound, 2.91 milliseconds). Primary ΔΔQTcI results were confirmed using Fridericia's formula for QTc. Categorical, morphological, and concentration–QTc analyses were consistent with the primary and secondary findings. There were no significant gender effects on ΔΔQTcI values. The most common adverse events were contact dermatitis and nausea (12.5% each) and dizziness (10.9%); adverse events were more frequent in the eluxadoline 1000 mg group. In conclusion, eluxadoline, at therapeutic or supratherapeutic doses, did not significantly prolong QT intervals, and was safe and generally well tolerated in this study population.
Pharmacologic and Pharmacodynamic Equivalence of 2 Formulations of Orlistat
Authors: Johnson Steven , Schwartz Susan M. 
Published: April 2018

AbstractWe sought to establish the bioequivalence of 2 weight‐loss aids: orlistat 27‐mg chewable tablet and orlistat 60‐mg capsule, measured pharmacodynamically as percentage fecal fat excretion. Two open‐label, single‐center, randomized, 3‐period, 3‐treatment crossover studies were conducted in adults with body mass index 25‐33 kg/m2. For each 9‐day treatment period, subjects received orlistat 27‐mg chewable tablet, 60‐mg capsule, or 120‐mg capsules (2 60‐mg capsules) 3 times daily; a 2‐day washout separated treatments. Primary bioequivalence analyses were based on 2 1‐sided tests of the 90% CI of the ratio of geometric means using log‐transformed data (study 1) and by the dose‐scale method to calculate bias‐corrected and accelerated 90% CI of relative bioavailability (f) using nontransformed data (study 2). Bioequivalence was established if 90% CIs fell within 0.80‐1.25. In total, 48 and 144 subjects were randomized in study 1 and study 2, respectively. Bioequivalence between the formulations was established in both studies: study 1 ratio of geometric means of percentage fecal fat excretion was 0.96 (2 1‐sided tests, 90% CI 0.87‐1.06); study 2‐point estimate of f was 1.09 (bias‐corrected and accelerated 90% CI 0.98‐1.22). Tolerability of the 27‐mg tablet was consistent with the 60‐mg capsule; mild gastrointestinal effects were most common.
Meningeal Whole Mount Preparation and Characterization of Neural Cells by Flow Cytometry
Authors: Louveau Antoine , Filiano Anthony J. , Kipnis Jonathan 
Published: April 2018

AbstractNeuroimmunologists aim to understand the interactions between the central nervous system and the immune system under both homeostatic and pathological conditions. The meninges, contrary to the brain parenchyma, are populated by numerous immune cells. Soluble factors produced by these cells are capable of diffusing into the brain parenchyma and influencing the brain cells within the parenchyma, including neurons. In this unit, we describe two protocols: the analysis of the meningeal compartment of rodents and the use flow cytometry to study the cells of the brain parenchyma (particularly neurons). © 2018 by John Wiley & Sons, Inc.
Introduction to Single‐Cell RNA Sequencing
Authors: Olsen Thale Kristin , Baryawno Ninib 
Published: April 2018

AbstractDuring the last decade, high‐throughput sequencing methods have revolutionized the entire field of biology. The opportunity to study entire transcriptomes in great detail using RNA sequencing (RNA‐seq) has fueled many important discoveries and is now a routine method in biomedical research. However, RNA‐seq is typically performed in “bulk,” and the data represent an average of gene expression patterns across thousands to millions of cells; this might obscure biologically relevant differences between cells. Single‐cell RNA‐seq (scRNA‐seq) represents an approach to overcome this problem. By isolating single cells, capturing their transcripts, and generating sequencing libraries in which the transcripts are mapped to individual cells, scRNA‐seq allows assessment of fundamental biological properties of cell populations and biological systems at unprecedented resolution. Here, we present the most common scRNA‐seq protocols in use today and the basics of data analysis and discuss factors that are important to consider before planning and designing an scRNA‐seq project. © 2018 by John Wiley & Sons, Inc.
Overview of Next‐Generation Sequencing Technologies
Authors: Slatko Barton E. , Gardner Andrew F. , Ausubel Frederick M. 
Published: April 2018

AbstractHigh throughput DNA sequencing methodology (next generation sequencing; NGS) has rapidly evolved over the past 15 years and new methods are continually being commercialized. As the technology develops, so do increases in the number of corresponding applications for basic and applied science. The purpose of this review is to provide a compendium of NGS methodologies and associated applications. Each brief discussion is followed by web links to the manufacturer and/or web‐based visualizations. Keyword searches, such as with Google, may also provide helpful internet links and information. © 2018 by John Wiley & Sons, Inc.
Making a Bacterial Thermophilic Enzyme in a Fungal Expression System
Authors: Nevalainen Helena , Bergquist Peter , Te'o Valentino Setoa Junior 
Published: April 2018

AbstractThis unit describes production of a bacterial thermophilic xylanase enzyme in an industrially exploited filamentous fungus, Trichoderma reesei. Successful expression of a gene of interest in a heterologous host involves front‐end design of the expression constructs using bioinformatics tools, making the constructs in the laboratory, and introducing them into the expression host. This is followed by synthesis and characterization of the gene product on a laboratory scale and optimization of the cultivation parameters in a controlled, scaled‐up fermentation. The thermophilic xylanase B (XynB) enzyme from the bacterium Dictyoglomus thermophilum discussed here can be easily purified by heat‐precipitation from the culture supernatant of the mesophilic host. A functional XynB can also be produced in Escherichia coli, but at a lower yield compared to that obtained in T. reesei. The protocol provided here can be adapted to various other proteins and filamentous fungal hosts. © 2018 by John Wiley & Sons, Inc.
Solubilization, Folding, and Purification of a Recombinant Peptidoglycan‐Associated Lipoprotein (PAL) Expressed in Escherichia coli
Authors: Santos Clelton A. , Souza Anete P. 
Published: April 2018

AbstractStudies aiming at heterologous expression of highly hydrophobic proteins, such as outer membrane proteins in general and peptidoglycan‐associated lipoprotein (PAL) in particular, are not trivial due to difficulties in obtaining recombinant protein in a soluble state, which is desired because it allows purification by traditional chromatographic methods. PAL is associated with the integrity of the cellular envelope in Gram‐negative bacteria and interacts strongly with the peptidoglycan layer. However, it is incorporated into inclusion bodies in studies focusing on its heterologous production. This protocol describes an efficient protein refolding method to solubilize and purify a recombinant PAL. Initially, recombinant PAL–enriched inclusion bodies obtained after the induction of PAL expression in Escherichia coli are treated with 8 M urea and then undergo buffer exchange via dialysis. Afterward, the soluble, recombinant PAL is purified using standard chromatographic methods. © 2018 by John Wiley & Sons, Inc.
Measurement of Homocitrulline, A Carbamylation‐derived Product, in Serum and Tissues by LC‐MS/MS
Authors: Jaisson Stéphane , Desmons Aurore , Doué Manon , Gorisse Laëtitia , Pietrement Christine , Gillery Philippe 
Published: April 2018

AbstractCarbamylation corresponds to the non‐enzymatic binding of isocyanic acid to protein amino groups and participates in protein molecular aging, characterized by the alteration of their structural and functional properties. Carbamylated proteins exert deleterious effects in vivo and are involved in the progression of various diseases, including atherosclerosis and chronic kidney disease. Therefore, there is a growing interest to evaluate the carbamylation rate of blood or tissue proteins, since carbamylation‐derived products (CDPs) constitute valuable biomarkers in these contexts. Homocitrulline, formed by isocyanic acid covalently attaches to the ε‐NH2 group of lysine residue side chain, is the most characteristic CDP. Sensitive and specific quantification of homocitrulline requires mass spectrometry‐based methods. This unit describes a liquid chromatography–tandem mass spectrometry (LC‐MS/MS) method for the quantification of homocitrulline, with special emphasis on pre‐analytical steps that allow quantification of total or protein‐bound homocitrulline in serum or tissue samples. © 2018 by John Wiley & Sons, Inc.
The limited contribution of large trees to annual biomass production in an old‐growth tropical forest
Authors: Ligot Gauthier , Gourlet‐Fleury Sylvie , Ouédraogo Dakis‐Yaoba , Morin Xavier , Bauwens Sébastien , Baya Fidele , Brostaux Yves , Doucet Jean‐Louis , Fayolle Adeline 
Published: April 2018

AbstractAlthough the importance of large trees regarding biodiversity and carbon stock in old‐growth forests is undeniable, their annual contribution to biomass production and carbon uptake remains poorly studied at the stand level. To clarify the role of large trees in biomass production, we used data of tree growth, mortality, and recruitment monitored during 20 yr in 10 4‐ha plots in a species‐rich tropical forest (Central African Republic). Using a random block design, three different silvicultural treatments, control, logged, and logged + thinned, were applied in the 10 plots. Annual biomass gains and losses were analyzed in relation to the relative biomass abundance of large trees and by tree size classes using a spatial bootstrap procedure. Although large trees had high individual growth rates and constituted a substantial amount of biomass, stand‐level biomass production decreased with the abundance of large trees in all treatments and plots. The contribution of large trees to annual stand‐level biomass production appeared limited in comparison to that of small trees. This pattern did not only originate from differences in abundance of small vs. large trees or differences in initial biomass stocks among tree size classes, but also from a reduced relative growth rate of large trees and a relatively constant mortality rate among tree size classes. In a context in which large trees are increasingly gaining attention as being a valuable and a key structural characteristic of natural forests, the present study brought key insights to better gauge the relatively limited role of large trees in annual stand‐level biomass production. In terms of carbon uptake, these results suggest, as already demonstrated, a low net carbon uptake of old‐growth forests in comparison to that of logged forests. Tropical forests that reach a successional stage with relatively high density of large trees progressively cease to be carbon sinks as large trees contribute sparsely or even negatively to the carbon uptake at the stand level.
Circular‐shaped microfluidic device to study the effect of shear stress on cellular orientation
Authors: Park Da Yeon , Kim Tae Hyeon , Lee Jong Min , Ahrberg Christian D. , Chung Bong Geun 
Published: April 2018

AbstractUnderstanding the effects of shear stress on mammalian cells is a crucial factor for understanding a number of biological processes and diseases. Here, we show the development of a circular‐shaped microfluidic device for the facile generation of shear stress gradients. With this microfluidic device, the effect of shear stress on orientation of human umbilical vein endothelial cells was studied. This microfluidic device, which enables to control the alignment of human umbilical vein endothelial cells within a microchannel, can be a valuable tool to mimic blood vessels.
Characterizing the fluid dynamics in the flow fields of cylindrical orbitally shaken bioreactors with different geometry sizes
Authors: Zhu Likuan , Han Wang , Song Boyan , Wang Zhenlong 
Published: April 2018

AbstractOrbitally shaken bioreactors (OSRs) are commonly used for the cultivation of mammalian cells in suspension. To aid the geometry designing and optimizing of OSRs, we conducted a three‐dimensional computational fluid dynamics (CFD) simulation to characterize the flow fields in a 10 L cylindrical OSR with different vessel diameters. The liquid wave shape captured by a camera experimentally validated the CFD models established for the cylindrical OSR. The geometry size effect on volumetric mass transfer coefficient (kLa) and hydromechanical stress was analyzed by varying the ratio of vessel diameter (d) to liquid height at static (hL), d/hL. The highest value of kLa about 30 h−1 was observed in the cylindrical vessel with the d/hL of 6.35. Moreover, the magnitudes of shear stress and energy dissipation rate in all the vessels tested were below their minimum values causing cells damage separately, which indicated that the hydromechanical‐stress environment in OSRs is suitable for cells cultivation in suspension. Finally, the CFD results suggested that the d/hL higher than 8.80 should not be adopted for the 10 L cylindrical OSR at the shaking speed of 180 rpm because the “out of phase” state probably will happen there.
Impact of nozzle operation on mass transfer in jet aerated loop reactors. Characterization and comparison to an aerated stirred tank reactor
Authors: Weber Sebastian , Schaepe Sebastian , Freyer Stephan , Kopf Michael‐Helmut , Dietzsch Christian 
Published: April 2018

AbstractThe impact of mass transfer on productivity can become a crucial aspect in the fermentative production of bulk chemicals. For highly aerobic bioprocesses the oxygen transfer rate
(OTR) and productivity are coupled. The achievable space time yields can often be correlated to the mass transfer performance of the respective bioreactor. The oxygen mass transfer capability of a jet aerated loop reactor is discussed in terms of the volumetric oxygen mass transfer coefficient kLa [h−1] and the energetic oxygen transfer efficiency E [kgO2 kW−1 h−1]. The jet aerated loop reactor (JLR) is compared to the frequently deployed aerated stirred tank reactor. In jet aerated reactors high local power densities in the mixing zone allow higher mass transfer rates, compared to aerated stirred tank reactors. When both reactors are operated at identical volumetric power input and aeration rates, local kLa values up to 1.5 times higher are possible with the JLR. High dispersion efficiencies in the JLR can be maintained even if the nozzle is supplied with pressurized gas. For increased oxygen demands (above 120 mmol L−1 h−1) improved energetic oxygen transfer efficiencies of up to 100 % were found for a JLR compared to an aerated stirred tank reactor operating with Rushton turbines.
The Critically Reflective Evaluator: Adult Education's Contributions to Evaluation for Social Justice
Authors: Archibald Thomas , Neubauer Leah C. , Brookfield Stephen D. 
Published: April 2018

AbstractIn this chapter, we suggest that current conversations about evaluative thinking would be enriched by the addition of insights from critical adult education rooted in critically reflective practice and critical theory. From our perspective situated in the praxis of adult education, we first review a sampling of literature on critical theory and critical reflection in evaluation, framed in terms of how evaluation theorists conceptualize the role of values and valuing in evaluation and how they envision the role of evaluation in society. Then, we explore some conceptual and practical overlaps between evaluative thinking, critical thinking, and critically reflective practice. Finally, we elucidate some implications of critical evaluative thinking for social justice evaluation, touching also on what implications this work might have even for those evaluators who do not perceive linkages and overlap between evaluative thinking and social justice. In doing so, we highlight new directions for the foundational training and continuing professional development of evaluation practitioners and scholars based on an examination of oneself and of one's paradigmatic assumptions through constant critical reflection, investigation, and action.
Argument as Conversation: Students Responding Through Writing to Significant Conversations Across Time and Place
Authors: VanDerHeide Jennifer , Juzwik Mary M. 
Published: April 2018

AbstractHow can social change become possible when secondary teachers approach argument as participation in ongoing conversations? The authors illustrate how—through writing, speaking, and reading to develop a letter to their principal arguing for a school policy change—secondary students participated in multiple conversational spheres across time and place. Although neither collaborative “letter writing to the principal” tasks nor “argument as conversation” stances are new, the approach presented is novel in its commitment to braiding oral and written language together within the framework of argument as conversation. This approach contrasts to more common practices of seeing conversation as preparation for writing. This approach is valuable not only for fostering students’ learning about processes, structures, and technical skills of argumentative writing but also, and more importantly, for experiencing what it feels like to become empowered disputants who affect social change in response to ongoing conversations that matter to students.
Three dimensional extrusion printing induces polymer molecule alignment and cell organization within engineered cartilage
Authors: Guo Ting , Ringel Julia P. , Lim Casey G. , Bracaglia Laura G. , Noshin Maeesha , Baker Hannah B. , Powell Douglas A. , Fisher John P. 
Published: April 2018

AbstractProper cell–material interactions are critical to remain cell function and thus successful tissue regeneration. Many fabrication processes have been developed to create microenvironments to control cell attachment and organization on a three‐dimensional (3D) scaffold. However, these approaches often involve heavy engineering and only the surface layer can be patterned. We found that 3D extrusion based printing at high temperature and pressure will result an aligned effect on the polymer molecules, and this molecular arrangement will further induce the cell alignment and different differentiation capacities. In particular, articular cartilage tissue is known to have zonal collagen fiber and cell orientation to support different functions, where collagen fibers and chondrocytes align parallel, randomly, and perpendicular, respectively, to the surface of the joint. Therefore, cell alignment was evaluated in a cartilage model in this study. We used small angle X‐ray scattering analysis to substantiate the polymer molecule alignment phenomenon. The cellular response was evaluated both in vitro and in vivo. Seeded mesenchymal stem cells (MSCs) showed different morphology and orientation on scaffolds, as a combined result of polymer molecule alignment and printed scaffold patterns. Gene expression results showed improved superficial zonal chondrogenic marker expression in parallel‐aligned group. The cell alignment was successfully maintained in the animal model after 7 days with distinct MSC morphology between the casted and parallel printed scaffolds. This 3D printing induced polymer and cell alignment will have a significant impact on developing scaffold with controlled cell–material interactions for complex tissue engineering while avoiding complicated surface treatment, and therefore provides new concept for effective tissue repairing in future clinical applications. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2190‐2199, 2018.
Comparing the reproducibility of commonly used magnetic resonance spectroscopy techniques to quantify cerebral glutathione
Authors: Wijtenburg S. Andrea , Near Jamie , Korenic Stephanie A. , Gaston Frank E. , Chen Hongji , Mikkelsen Mark , Chen Shuo , Kochunov Peter , Hong L. Elliot , Rowland Laura M. 
Published: April 2018

BackgroundCerebral glutathione (GSH), a marker of oxidative stress, has been quantified in neurodegenerative diseases and psychiatric disorders using proton magnetic resonance spectroscopy (MRS). Using a reproducible MRS technique is important, as it minimizes the impact of measurement technique variability on the study results and ensures that other studies can replicate the results.HypothesisWe hypothesized that very short echo time (TE) acquisitions would have comparable reproducibility to a long TE MEGA‐PRESS acquisition, and that the short TE PRESS acquisition would have the poorest reproducibility.Study TypeProspective.Subjects/PhantomsTen healthy adults were scanned during two visits, and six metabolite phantoms containing varying concentrations of GSH and metabolites with resonances that overlap with GSH were scanned once.Field Strength/SequenceAt 3T we acquired MRS data using four different sequences: PRESS, SPECIAL, PR‐STEAM, and MEGA‐PRESS.AssessmentReproducibility of each MRS sequence across two visits was assessed.Statistical TestsMean coefficients of variation (CV) and mean absolute difference (AD) were used to assess reproducibility. Linear regressions were performed on data collected from phantoms to examine the agreement between known and quantified levels of GSH.ResultsOf the four techniques, PR‐STEAM had the lowest mean CV and AD (5.4% and 7.5%, respectively), implying excellent reproducibility, followed closely by PRESS (5.8% and 8.2%) and SPECIAL (8.0 and 10.1%), and finally by MEGA‐PRESS (13.5% and 17.1%). Phantom data revealed excellent fits (R2 ≥ 0.98 or higher) using all methods.Data ConclusionOur data suggest that GSH can be quantified reproducibly without the use of spectral editing.Level of Evidence: 2Technical Efficacy: Stage 2J. Magn. Reson. Imaging 2019;49:176–183.
Detection of a new KCNQ1 frameshift mutation associated with Jervell and Lange‐Nielsen syndrome in 2 Iranian families
Authors: Amirian Azam , Zafari Zahra , Dalili Mohammad , Saber Siamak , Karimipoor Morteza , Dabbagh Bagheri Samira , Fazelifar Amir Farjam , Zeinali Sirous 
Published: April 2018

AbstractJervell‐Lange Nielsen syndrome (JLNS) with autosomal recessive inheritance is a congenital cardiovascular disorder characterized by prolongation of QT interval on the ECG and deafness. We have performed molecular investigation by haplotype analysis and DNA Sanger sequencing in 2 unrelated Iranian families with a history of syncope. Mutational screening of KCNQ1 gene revealed the novel homozygous frameshift mutation c.733‐734delGG (p.G245Rfs*39) in 2 obviously unrelated cases of JLNS which is probably a founder mutation in Iran. The novel mutation detected in this study is the first time reported among Iranian population and will be beneficial in the tribe and region‐specific cascade screening of LQTS in Iran.