bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection of Tyrosine Sulfation on Proteins
Authors: Kanan Yogita , Al Ubaidi Muayyad R. 
Published: April 2015

AbstractTyrosine sulfation is a post‐translational modification (PTM) where a sulfate group is added to a tyrosine moiety. This PTM is responsible for strengthening interaction between proteins. One of the drawbacks of studying this PTM is the lack of an antibody that can detect all tyrosine‐sulfated proteins. In addition, due to the labile nature of the tyrosine sulfate, other techniques such as mass spectrometry cannot be used to study this PTM unless special modification procedures are used. This requires considerable skill and knowledge of mass spectrometry. This unit describes an in vitro technique that can be used to study tyrosine‐sulfated proteins by radiolabeling the recombinant protein. The protein is then subject to barium hydroxide hydrolysis and thin‐layer electrophoresis (TLE). Co‐localization of radioactive tyrosine‐sulfate with nonradioactive tyrosine sulfate standard spiked in before TLE analysis identifies a protein as tyrosine‐sulfated protein. The advantage of this technique is that, it identifies all tyrosine‐sulfated proteins without any bias and is the only technique that identifies the tyrosine sulfate residues in the protein. © 2015 by John Wiley & Sons, Inc.
Preparing Protein Extracts for Quantitative Two‐Dimensional Gel Comparison
Authors: Lelong Cécile , Rabilloud Thierry 
Published: April 2015

AbstractThis unit describes basic protocols for efficient and reproducible protein solubilization from a variety of biological samples, including cultured animal cells and tissues, plant cells and tissues, bacteria, nuclei, other subcellular organelles, plasma, serum, and other biological fluids. The optimized extraction process is strongly sample dependent and cannot be described for every type of sample. Instead, typical protocols are provided as general guidelines and illustrate good starting points for optimization of sample preparation. These solubilization procedures take into account the constraints imposed by two‐dimensional electrophoresis and are thus well suited for proteomic approaches. © 2015 by John Wiley & Sons, Inc.
Latent Sensitization: A Model for Stress‐Sensitive Chronic Pain
Authors: Marvizon Juan Carlos , Walwyn Wendy , Minasyan Ani , Chen Wenling , Taylor Bradley K. 
Published: April 2015

AbstractLatent sensitization is a rodent model of chronic pain that reproduces both its episodic nature and its sensitivity to stress. It is triggered by a wide variety of injuries ranging from injection of inflammatory agents to nerve damage. It follows a characteristic time course in which a hyperalgesic phase is followed by a phase of remission. The hyperalgesic phase lasts between a few days to several months, depending on the triggering injury. Injection of μ‐opioid receptor inverse agonists (e.g., naloxone or naltrexone) during the remission phase induces reinstatement of hyperalgesia. This indicates that the remission phase does not represent a return to the normal state, but rather an altered state in which hyperalgesia is masked by constitutive activity of opioid receptors. Importantly, stress also triggers reinstatement. Here we describe in detail procedures for inducing and following latent sensitization in its different phases in rats and mice. © 2015 by John Wiley & Sons, Inc.
Purification of Recombinant Proteins from Cultured Mammalian Cells by HaloTag Technology
Authors: Ohana Rachel Friedman , Hurst Robin 
Published: April 2015

AbstractCultured mammalian cells provide an environment ideal for producing functional recombinant mammalian proteins. However, low expression levels of recombinant proteins present a challenge for their detection and purification. This unit will focus on HaloTag, a protein fusion tag designed to bind selectively and covalently to a chloroalkane ligand that may be attached to a variety of functional groups, allowing both protein detection and immobilization. Detection of HaloTag‐fusion protein is achieved through binding to a fluorescent chloroalkane ligand, enabling rapid optimization of expression levels. HaloTag‐based purification uses covalent capture of the HaloTag fusion onto HaloLink resin coupled with proteolytic cleavage to release the protein of interest from the resin. Covalent binding provides efficient protein capture regardless of expression level and eliminates protein loss during washes of the resin and as a result, offers significant improvements in protein recovery and purity over traditional non‐covalent approaches. © 2015 by John Wiley & Sons, Inc.
Measurement of Bacterial Ingestion and Killing by Macrophages
Authors: Drevets Douglas A. , Canono Beth P. , Campbell Priscilla A. 
Published: April 2015

AbstractThis unit presents assays that allow accurate measurement of phagocytosis and killing of bacteria by macrophages. The first basic protocol describes how to measure the ability of macrophages to ingest bacteria. Importantly, because macrophage phagocytosis entails separate binding and internalization steps, assays are described here that will also determine the extent to which bacteria bound to the macrophage are in fact internalized. Two effective methods to do this are described in alternate protocols. Both of these alternate protocols rely on enumeration of differentially labeled bacteria by fluorescence microscopy to distinguish intracellular from extracellular bacteria. The unit also presents two protocols to measure the ability of a macrophage to kill bacteria it has internalized. The second basic is a straightforward assay in which bacterial colonies are enumerated before and after a killing period. Bactericidal activity is evidenced by reduced CFU bacteria on agar plates. Because it is critical to remove residual extracellular organisms, the protocol presents two alternative steps to accomplish this: a washing procedure and a more stringent method in which cells are sedimented through sucrose. An alternate protocol describes a way to measure bacterial viability based on bacterial metabolism, in which the ability of bacterial dehydrogenases to mediate the reduction of a tetrazolium salt to purple formazan is monitored by measuring absorbance spectrophotometrically. © 2015 by John Wiley & Sons, Inc.
High‐Sensitivity Detection of PNH Red Blood Cells, Red Cell Precursors, and White Blood Cells
Authors: Sutherland D. Robert , Illingworth Andrea , Keeney Michael , Richards Stephen J. 
Published: April 2015

AbstractFlow cytometry is the method of choice to ‘diagnose’ paroxysmal nocturnal hemoglobinuria (PNH) and has led to improved patient management. Most laboratories have limited experience with PNH testing, and many different flow approaches are used. Careful selection and validation of antibody conjugates has allowed the development of reagent cocktails suitable for detection of PNH RBCs, CD71+ reticulocytes, and WBCs in clinical/sub‐clinical PNH samples. A CD235a‐FITC/CD59‐PE assay was developed capable of detecting Type III PNH RBCs at 0.01% sensitivity. A protocol targeting immature CD71+ RBCs can detect PNH reticulocytes at similar sensitivity. Four‐color FLAER‐based neutrophil and monocyte assays were developed to detect PNH phenotypes at a level of 0.01% and 0.04% sensitivity, respectively. For instrumentation with five or more PMTs, a single‐tube 5‐color FLAER/CD157‐based assay to simultaneously detect PNH neutrophils and monocytes is described. Using these standardized approaches, results have demonstrated good intra‐ and inter‐laboratory performance characteristics even in laboratories with little prior experience performing PNH testing. © 2015 by John Wiley & Sons, Inc.
Flow Cytometry of Murine Spermatocytes
Authors: Gaysinskaya Valeriya , Bortvin Alex 
Published: April 2015

AbstractProtocols for purification of murine male germ cells by FACS based on Hoechst 33342 (Ho342) dye staining have been reported and optimized. However, the protocols are often challenging to follow, partly due to difficulties related to sample preparation, instrument parameters, data display, and selection strategies. In addition, troubleshooting of flow cytometry experiments usually requires some fluency in technical principles and instrument specifications and settings. This unit describes setup and procedures for analysis and sorting of male meiotic prophase I (MPI) cells and other germ cells. Included are procedures that guide data acquisition, display, gating, and back‐gating critical for optimal data visualization and cell sorting. Additionally, a flow cytometry analysis of spermatogenesis‐defective testis is provided to illustrate the applicability of the technique to the characterization and purification of cells from mutant testis. © 2015 by John Wiley & Sons, Inc.
Uncovering the geodetic signature of silent slip through repeating earthquakes
Authors: Frank William B. , Radiguet Mathilde , Rousset Baptiste , Shapiro Nikolaï M. , Husker Allen L. , Kostoglodov Vladimir , Cotte Nathalie , Campillo Michel 
Published: April 2015

AbstractSlow transient slip that releases stress along the deep roots of plate interfaces is most often observed on regional GPS networks installed at the surface. The detection of slow slip is not trivial if the dislocation along the fault at depth does not generate a geodetic signal greater than the observational noise level. Instead of the typical workflow of comparing independently gathered seismic and geodetic observations to study slow slip, we use repeating low‐frequency earthquakes to reveal a previously unobserved slow slip event. By aligning GPS time series with episodes of low‐frequency earthquake activity and stacking, we identify a repeating transient slip event that generates a displacement at the surface that is hidden under noise prior to stacking. Our results suggest that the geodetic investigation of transient slip guided by seismological information is essential in exploring the spectrum of fault slip.
Numerical study of anomalous absorption of O mode waves on magnetic field‐aligned striations
Authors: Eliasson B. , Papadopoulos K. 
Published: April 2015

AbstractSimple expressions that describe mode conversion and anomalous absorption of ordinary (O) mode waves injected at angles between the vertical and magnetic zenith to upper hybrid (UH) oscillations in the presence of field‐aligned density striations are presented. The absorption takes place in a region above the UH resonance layer where the striations allow trapped eigenmodes, leading to excitation of large‐amplitude UH waves. The derivation of the expressions is guided by dimensional analysis and numerical simulations. The results are relevant in interpreting high‐latitude heating experiments where anomalous absorption due to striations plays a crucial role.
Mechanics of Contacting Surfaces
Authors: Rahnejat Homer , Johns‐Rahnejat Patricia Margaret 
Published: April 2015

Abstract
Many load bearing or power transmitting conjunctions in engine and drivetrain are counterforming concentrated contacts. The mechanics of contact is described by small strain localized deformation, described by Hertzian contact mechanics. Usually, a thin film of lubricant exists in contacts such as in the cam–follower pair, ball and rolling element bearings, and meshing gear teeth pairs. The regime of lubrication is invariably mixed elastohydrodynamics. This chapter provides an introduction to the fundamentals of contact mechanics, Hertzian theory, and elastohydrodynamic regime of lubrication. It also highlights the mechanisms of viscous and boundary friction in concentrated contacts.
Engine/Transmission Matching
Authors: Mariotti Gabriele Virzi' 
Published: April 2015

AbstractThis chapter deals with the forecasting of vehicle performance on the road, according to the installed engine. It begins with Section 1, which shows the calculation of the resistance to motion, that is, the forces that the vehicle must overcome to advance on flat roads and uphill. The calculation excludes accidental resistance in curves because it is not essential for predicting the behavior of the vehicle on the road. Section 2 begins with the power characteristic, the forecast by traction–speed and power–speed diagrams, calculation of gear ratios, and then passing to the limits to the estimated acceleration, feature consumption, and distance. This chapter concludes with a section on the drivability of the vehicle, and sets out the advantages of four‐wheel‐drive vehicles.
Automotive Diesel Engine Development Trends
Authors: Tomazic Dean , Koerfer Thomas , Pischinger Stefan 
Published: April 2015

Abstract
Since the invention of the diesel engine by Rudolf Diesel, the diesel technology has evolved to a point where this engine concept can be found in small engines for power generation to automotive applications up to large trucks, large bore ships/locomotives, and other industrial engine applications. With its superior fuel consumption over any other internal reciprocating combustion engine, it has become a prime choice for passenger car and light‐duty trucks all over the world. Despite the introduction of continuously more stringent emissions legislation, the development of new innovative technologies in the area of emissions, controls, sensors, and materials has allowed the diesel engine to remain competitive and maintain its position in the marketplace.
This chapter highlights the historic background and current and future technologies for automotive diesel engine applications focusing on all important aspects such as emissions, fuel economy, fuels, design and materials, combustion system configurations, and fuel injection systems and their relevance to meeting future targets.
Cellular Mobile Networks
Authors: Ramme Friedhelm , Fiege Gero , Rembarz René 
Published: April 2015

AbstractBeing connected while on the move has become a common customer expectation. This expectation goes hand in hand with a change in the observed user behavior. Nowadays, apps and connected devices have become part of people's everyday life. Cellular networks are assumed to provide the required connectivity whenever and wherever needed, which is a big change compared to the situation just two decades ago. A further and society‐impacting trend, with major economic and environmental implications, is caused by the ever‐growing demand for personal and goods mobilities. Steadily raising road traffic levels, particularly in developed or rapidly developing counties, are spurring demands for real‐time traffic information, safety applications, and a broad set of connected car services. Hence, the embedded expectation on ubiquitous cellular connectivity is becoming increasingly important for the mobile society and the just‐in‐time economy landscape. The changing perception on deployed cellular networks and recent technology advancements are leading to difficult arguments for additional large‐scale investments in a dedicated vehicle communication infrastructure for certain driver or passenger services.Either way, we do not discuss anymore whether cars will become connected in the future—they will! The question is rather: what is the best option to realize tailored connectivity for automotive applications from a technology perspective, considering operational, economic, and timing conditions?This chapter provides some basic information about cellular communication systems, required to properly address those questions. Major trends in cellular communications and resulting impacts to business and society are discussed in Section 2. The various generations of cellular wireless communication systems, their key characteristics and prime network elements are introduced in Section 3. Section 4 illustrates selected use case procedures and how core network components interact with each other in an integrated system design. Special attention is put on advanced scenarios with particular relevance for automotive communication. Finally, Section 5 provides a short introduction on the standards and regulatory frameworks related to cellular communication.
A Layered P2‐ and O3‐Type Composite as a High‐Energy Cathode for Rechargeable Sodium‐Ion Batteries
Authors: Guo Shaohua , Liu Pan , Yu Haijun , Zhu Yanbei , Chen Mingwei , Ishida Masayoshi , Zhou Haoshen 
Published: April 2015

AbstractA layered composite with P2 and O3 integration is proposed toward a sodium‐ion battery with high energy density and long cycle life. The integration of P2 and O3 structures in this layered oxide is clearly characterized by XRD refinement, SAED and HAADF and ABF‐STEM at atomic resolution. The biphase synergy in this layered P2+O3 composite is well established during the electrochemical reaction. This layered composite can deliver a high reversible capacity with the largest energy density of 640 mAh g−1, and it also presents good capacity retention over 150 times of sodium extraction and insertion.
Palladium‐Catalyzed Hydrocarboxylation of Alkynes with Formic Acid
Authors: Hou Jing , Xie Jian‐Hua , Zhou Qi‐Lin 
Published: April 2015

AbstractA palladium‐catalyzed hydrocarboxylation of alkynes with formic acid has been developed. The method provides acrylic acid and derivatives in good yields with high regioselectivity without the need to handle toxic CO gas.
Synthesis of a new Schiff‐base complex based on Cu and its inference in PVC‐based membrane as an electroactive material
Authors: Swami Babu Lal , Ikram Saiqa 
Published: April 2015

AbstractA novel Schiff‐base complex based on copper [Cu(L1)2] derived from ligand 2‐[(E)‐(2‐hydroxy naphthalen‐1‐yl)‐methylidene amino] isoindoline‐1, 3‐dione (L1) is reported by the reaction of N‐aminophthalimide with 2‐hydroxy‐1‐naphthaldehyde in presence of ethanol and hydrazine hydrate and characterized by Fourier transform infrared spectroscopy (FT‐IR), Proton NMR (1HNMR) and UV–vis spectroscopic techniques. The synthesized Schiff‐base complex [Cu(L1)2] was employed as an electroactive material within poly (vinyl chloride) matrix in order to obtain a membrane by using di‐butyl‐phthalate as plasticizer which demonstrated good transferring of metal ions proposing its application as an ion selective material. © 2015 Curtin University of Technology and John Wiley & Sons, Ltd.
Processing and characterization of bio‐based poly (hydroxyalkanoate)/poly(amide) blends: Improved flexibility and impact resistance of PHA‐based plastics
Authors: Yang Shengzhe , Madbouly Samy A. , Schrader James A. , Grewell David , Kessler Michael R. , Graves William R. 
Published: April 2015

ABSTRACTOne of the most significant limitations to widespread industrial implementation of emerging bioplastics such as poly(lactic acid) and poly(hydroxyalkanoate) (PHA) is that they do not match the flexibility and impact resistance of petroleum‐based plastics like poly(propylene) or high‐density poly(ethylene). The basic goal of this research is to identify alternative, affordable, sustainable, biodegradable materials that can replace petroleum‐based polymers in a wide range of industrial applications, with an emphasis on providing a solution for increasing the flexibility of PHA to a level that makes it a superior material for bioplastic nursery‐crop containers. A series of bio‐based PHA/poly(amide) (PA) blends with different concentrations were mechanically melt processed using a twin‐screw extruder and evaluated for physical characteristics. The effects of blending on viscoelastic properties were investigated using small‐amplitude oscillatory shear flow experiments to model the physical character as a function of blend composition and angular frequency. The mechanical, thermal, and morphological properties of the blends were investigated using dynamic mechanical analysis, differential scanning calorimetry, thermogravimetric analysis, scanning electron microscopy, and tensile tests. The complex viscosity of the blends increased significantly with increasing concentration of PHA and reached a maximum value for 80 wt % PHA blend. In addition, the tensile strength of the blends increased markedly as the content of PHA increased. For blends containing PA at >50 wt %, samples failed only after a very large elongation (up to 465%) without significant decrease in tensile strength. The particle size significantly increased and the blends became more brittle with increasing concentration of PHA. In addition, the concentration of the PA had a substantial effect on the glass relaxation temperature of the resulting blends. Our results demonstrate that the thermomechanical and rheological properties of PHA/PA blends can be tailored for specific applications, and that blends of PHA/PA can fulfill the mechanical properties required for flexible, impact‐resistant bio‐based nursery‐crop containers. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 42209.
Processing and properties of phthalic anhydride modified soy protein/glycerol plasticized soy protein composite films
Authors: Guo Gaiping , Zhang Chen , Du Zhongjie , Zou Wei , Xiang Aimin , Li Hangquan 
Published: April 2015

ABSTRACTPhthalic anhydride modified soy protein (PAS)/glycerol plasticized soy protein (GPS) composite films were fabricated by using extrusion and compression‐molding. Modified with phthalic anhydride, the soy protein lost its thermoplastic ability and was used as a filler to reinforce the GPS matrix. Fourier transform infrared spectra, optical transmittance, scanning electron microscope, mechanical tests, water resistance tests, as well as thermo‐gravimetric analysis were carried out to investigate the structure and properties of PAS and the plastic composites. The similar chemical structure of PAS and GPS led to compatibility of the two components resulting in high transparency and enhanced tensile properties of the composites. The water resistance of GPS was also improved by the incorporation of PAS. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 42221.
Methods of long chain branching detection in PE by triple‐detector gel permeation chromatography
Authors: Pathaweeisariyakul Thipphaya , Narkchamnan Kanyanut , Thitisuk Boonyakeat , Yau Wallace 
Published: April 2015

ABSTRACTLong chain branching (LCB) in polyethylene is one of the key microstructures that controls processing and final properties. Gel permeation chromatography (GPC) with viscometer (IV) and/or light scattering (LS) has been intensely used to quantify LCB. The widespread method to quantify LCB from GPC with IV or LS is the method of LCB frequency (LCBf) based on the Zimm–Stockmayer (ZS) random branching model. In this work, the conventional approach was compared with the recently developed method, called gpcBR. The comparison of the sensitivity of both methods is made on highly branched polymer, that is, various grades of commercial LDPE and also on polymer with very low level of LCB, that is, a commercial HDPE with no LCB, converted into several branched test samples of gradually increasing LCB by multiple extrusion. Finally, the linkages of LCB quantities from both methods to the rheological data and processing properties are illustrated. The new gpcBR index can access lower LCB level and shows obviously better relationship with both rheological data and processing properties than LCBf from the conventional ZS model. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 42222.