bluuebunny/crossref_metadata_2025 huggingface dataset:
Lyn regulates inflammatory responses in Klebsiella pneumoniae infection via the p38/NF‐κB pathway
Authors: Li Xuefeng , Zhou Xikun , Ye Yan , Li Yi , Li Jiaxin , Privratsky Breanna , Wu Erxi , Gao Hongwei , Huang Canhua , Wu Min
Published: November 2013

Klebsiella pneumoniae (Kp) is one of the most common pathogens in nosocomial infections and is becoming increasingly multidrug resistant. However, the underlying molecular pathogenesis of this bacterium remains elusive, limiting the therapeutic options. Understanding the mechanism of its pathogenesis may facilitate the development of anti‐bacterial therapeutics. Here, we show that Lyn, a pleiotropic Src tyrosine kinase, is involved in host defense against Kp by regulating phagocytosis process and simultaneously downregulating inflammatory responses. Using acute infection mouse models, we observed that lyn−/− mice were more susceptible to Kp with increased mortality and severe lung injury compared with WT mice. Kp infected‐lyn−/− mice exhibited elevated inflammatory cytokines (IL‐6 and TNF‐α), and increased superoxide in the lung and other organs. In addition, the phosphorylation of p38 and NF‐κB p65 subunit increased markedly in response to Kp infection in lyn−/− mice. We also demonstrated that the translocation of p65 from cytoplasm to nuclei increased in cultured murine lung epithelial cells by Lyn siRNA knockdown. Furthermore, lipid rafts clustered with activated Lyn and accumulated in the site of Kp invasion. Taken together, these findings revealed that Lyn may participate in host defense against Kp infection through the negative modulation of inflammatory cytokines.
Pd17In4Se4, a Metal‐Rich Palladium–Indium Selenide with an Open‐Framework Structure
Authors: Zakharova Elena Yu. , Churakov Andrei V. , Doert Thomas , Kuznetsov Alexey N.
Published: November 2013

A new metal‐rich palladium–indium selenide, Pd17In4Se4, was synthesized in the shape of needlelike crystals by annealing a mixture of the elements in a 7:2:1 ratio, respectively, at 750 °C for 7–10 d with subsequent cooling to room temperature. Its crystal structure was determined from single‐crystal diffraction data, with the composition confirmed by energy‐dispersive spectroscopy (EDS). The compound crystallizes in a new structure type [tetragonal,I4/m,a= 14.3104(7) Å,c= 3.9641(4) Å,Z= 2]. Its main structural units are indium‐centered [Pd12In] cuboctahedra of the Cu3Au type. The key feature that differentiates the structure of Pd17In4Se4from any known compound based on the Pd–In intermetallic motif is the terminal role of selenium atoms that results in the formation of 1D channels along thecaxis. Electronic structure calculations on the DFT/full‐potential linearized augmented‐plane wave (FP‐LAPW) level predict metallic properties and Pauli‐like paramagnetic behavior for the new compound. The electron localization function (ELF) topological analysis reveals the key role of selenium lone pairs in the formation of the channels in the structure and shows the multicentered nature of the bonding in the [Pd12In] fragments.
On‐Demand Selection of the Reaction Path from Imino Diels–Alder to Ene‐Type Cyclization: Synthesis of Epiminopyrimido[4,5‐b]azepines
Authors: Zhang Yuewei , Zhu Yue , Zheng Lianyou , Zhuo Lian‐Gang , Yang Fengzhi , Dang Qun , Yu Zhi‐Xiang , Bai Xu
Published: November 2013

Controlling the mode of reaction of a reactive intermediate such as an imine or iminium ion should enable the on‐demand selection of the final products from the same starting materials. The successful execution of such a strategy will reduce the time required to prepare diverse scaffolds. The imines derived from 4‐(allylamino)pyrimidine‐5‐carbaldehydes and anilines undergo Diels–Alder reactions to give pyrimido[4,5‐h][1,6]naphthyridines in high yields. A complete switch from the intramolecular aza‐Diels–Alder (IADA) path to an ene‐type cyclization reaction was achieved by simply adjusting the reaction conditions (amount of acid catalyst, solvent, and temperature). This newly introduced ene‐type cyclization reaction was used to prepare a series of epiminopyrimido[4,5‐b]azepines. To gain insight into the mechanism of the two reaction pathways, a DFT study was carried out. Theoretical calculations showed that under acidic conditions an iminium intermediate favors the low‐energy IADA pathway, which proceeds in a [4+ + 2] fashion. When acid is absent, the neutral imine intermediate favors the thermal ene‐type cyclization reaction, which takes place by transfer of an allylic proton from the allylic amine to imine, followed by a barrierless nucleophilic addition process between the in‐situ‐generated anionic allylic amine and iminium ion. Amine addition to the alkene finally gives the epiminopyrimido[4,5‐b]azepines.
Copper‐Catalyzed N‐ and O‐Arylation of Amides: Alternative Approaches to 3,4‐Dihydroquinolin‐2‐ones, Quinolin‐2‐ones, and 12H‐Chromeno[2,3‐b]quinolin‐12‐ones
Authors: Zhang Xinying , Guo Xiaojie , Fang Liangliang , Song Yunping , Fan Xuesen
Published: November 2013

The efficient syntheses of 3‐cyanoquinolin‐2‐ones, 3‐(2‐bromobenzyl)‐3‐cyano‐3,4‐dihydroquinolin‐2‐ones, 3‐(2‐bromobenzyl)quinolin‐2‐ones, and 12H‐chromeno[2,3‐b]quinolin‐12‐ones through copper‐catalyzed N‐ and O‐arylation of amides by using readily available 2‐bromobenzyl bromides and cyanoacetamides as starting materials were developed.
Highly Enantioselective Biginelli Reaction Catalyzed by Double Axially Chiral Bisphosphorylimides
Authors: An Dong , Fan Yan‐Sen , Gao Yang , Zhu Zi‐Qian , Zheng Liang‐Yu , Zhang Suo‐Qin
Published: November 2013

Double axially chiral bisphosphorylimides have been used as catalysts in enantioselective Biginelli reactions. The three‐component reaction of aromatic aldehydes, thiourea, and ethyl acetoacetate took place by using 5 mol‐% catalyst in ethyl acetate at 50 °C. A series of chiral dihydropyrimidinethiones (DHPMs) were obtained in high yields (up to 97 %) with good to high enantioselectivities (up to 96 % ee) in only 12 hours.
Topographic locations and size of earthquake‐ and typhoon‐generated landslides, Tachia River, Taiwan
Authors: Huang Alice Y.‐L. , Montgomery David R.
Published: November 2013

Analysis of mapped landslide locations using a high‐resolution (5‐m grid) digital elevation model (DEM) in the Tachia River basin, Taiwan, finds distinct differences in the topographic locations and size of landslides during the 1999 Chi‐Chi earthquake and the 2001 Toraji typhoon. Our analysis supports Densmore and Hovius' hypothesis that earthquake‐induced landslides cluster near ridgetops due to topographic amplification of ground shaking, and that typhoon‐induced landslides occur with greater frequency lower on slopes. In addition, the differing topographic locations of seismically‐induced and subsequent typhoon‐induced landslides shows no evidence of residual post‐earthquake influences on landslides during typhoon Toraji previously hypothesized for drainage basins closer to the earthquake epicenter. Our results support the interpretation that in this tectonically active landscape, seismically‐induced landslides help shatter and erode ridgetops but typhoon‐triggered landslides concentrate erosion farther downslope, with the combination acting to more uniformly lower upland terrain than either process does individually. Copyright © 2013 John Wiley & Sons, Ltd.
The influence of bedrock orientation on the landscape evolution, surface morphology and denudation (10Be) at the Niesen, Switzerland
Authors: Chittenden Hannah , Delunel Romain , Schlunegger Fritz , Akçar Naki , Kubik Peter
Published: November 2013

Landscape evolution and surface morphology in mountainous settings are a function of the relative importance between sediment transport processes acting on hillslopes and in channels, modulated by climate variables. The Niesen nappe in the Swiss Penninic Prealps presents a unique setting in which opposite facing flanks host basins underlain by identical lithologies, but contrasting litho‐tectonic architectures where lithologies either dip parallel to the topographic slope or in the opposite direction (i.e. dip slope and non‐dip slope). The north‐western facing Diemtigen flank represents such a dip slope situation and is characterized by a gentle topography, low hillslope gradients, poorly dissected channels, and it hosts large landslides. In contrast, the south‐eastern facing Frutigen side can be described as non‐dip slope flank with deeply incised bedrock channels, high mean hillslope gradients and high relief topography. Results from morphometric analysis reveal that noticeable differences in morphometric parameters can be related to the contrasts in the relative importance of the internal hillslope‐channel system between both valley flanks. While the contrasting dip‐orientations of the underlying flysch bedrock has promoted hillslope and channelized processes to contrasting extents and particularly the occurrence of large landslides on the dip slope flank, the flank averaged beryllium‐10 (10Be)‐derived denudation rates are very similar and range between 0.20 and 0.26 mm yr−1. In addition, our denudation rates offer no direct relationship to basin's slope, area, steepness or concavity index, but reveal a positive correlation to mean basin elevation that we interpret as having been controlled by climatically driven factors such as frost‐induced processes and orographic precipitation. Our findings illustrate that while the landscape properties in this part of the northern Alpine border can mainly be related to the tectonic architecture of the underlying bedrock, the denudation rates have a strong orographic control through elevation dependent mean annual temperature and precipitation. Copyright © 2013 John Wiley & Sons, Ltd.
Environmental Regulation: BACT Determination for GHG Permitting Requires Care
Authors: Hughes Deanne Dutton
Published: November 2013

When contemplating significant operational or physical changes at a facility, an owner must carefully consider the impact on a facility's air quality permit. Often, the air quality construction permitting process becomes a critical path item for meeting project objectives.
Electric Regulation: Profitability of CO2 Emissions Depends on Available Markets
Authors: Bowden Nicholas S.
Published: November 2013

The White House's June 2013 Climate Action Plan stated, in part, “the Obama Administration is putting in place tough new rules to cut carbon pollution—just like we have for other toxins like mercury and arsenic—so we protect the health of our children and move our economy toward American‐made clean energy sources that will create good jobs and lower home energy bills.” In April 2012, the United States Environmental Protection Agency (EPA) proposed carbon dioxide (CO2) emissions standards for future electric‐generating units. After receiving more than 2.5 million sets of public comments, this first proposal was rescinded and replaced with a revised proposal on September 20, 2013. However, the proposal itself claims that it will do little to nothing to reduce CO2 emissions, as the standards only apply to generating units constructed in the future.
Implementing Performance Appraisal: Exploring the Employee Experience
Authors: Farndale Elaine , Kelliher Clare
Published: November 2013

Line managers play an important role as implementers of performance appraisal, enacting procedures designed by the HR function. However, the actual employee experience of these procedures (which may differ from how they were intended or enacted) in terms of perceptions of justice in the process is likely to have consequences for levels of organizational commitment. Furthermore, based on signaling theory, the broader organizational climate, measured here in terms of the level of trust employees have in the senior management, sets the context in which this experience takes shape. Presenting multilevel analysis of 4,422 employees across 22 business units, we show that organizational units with high trust in senior management have both higher levels of commitment, and show a stronger link between employee perceptions of fair treatment by their line manager during performance appraisal, and organizational commitment. This provides initial evidence that the impact of line manager actions is important for employee‐level outcomes but is also constrained by the organizational climate. © 2013 Wiley Periodicals, Inc.
Employee Perceptions of Line Management Performance: Applying the AMO Theory to Explain the Effectiveness of Line Managers' HRM Implementation
Authors: Bos‐Nehles Anna C. , Van Riemsdijk Maarten J. , Kees Looise Jan
Published: November 2013

Line managers are today seen as increasingly important in effectively implementing HRM practices. Based on the Ability‐Motivation‐Opportunity (AMO) theory, we predict that line managers' performance in this regard will depend on their ability to apply HRM practices, and that their motivation and the opportunity provided will enhance this effect. Through a survey of 174 line managers and 1,065 of their direct subordinates in two organizations, we found ability to be the best predictor of a line manager's HRM performance. Motivation did not moderate the effect of ability on performance as predicted, and our results suggest this relationship needs further attention. Opportunity did enhance the effect of ability on HRM implementation effectiveness. Therefore, HRM departments should aim to enhance line managers' abilities and provide adequate opportunities for them to carry out the expected HRM duties. © 2013 Wiley Periodicals, Inc.
The Effect of Strategic Human Resource Management on Organizational Performance: The Mediating Role of High‐Performance Human Resource Practices
Authors: Mitchell Rebecca , Obeidat Shatha , Bray Mark
Published: November 2013

This study investigates the impact of a strategic role for human resource management (HRM) on organizational financial performance. Survey data from 118 financial and manufacturing organizations in Jordan is analyzed to investigate a moderated mediated pathway between a strategic role for HRM and organizational financial performance. The first finding, that high‐performance human resource practices (HPHRPs) mediate the relationship between the HR strategic role and performance, suggests that the position of the HR professional as a strategic partner enhances the legitimacy of HR initiatives, as well as facilitating the resourcing and support for their implementation. The second finding, that line management devolvement moderates the relationship between the HR strategic role and HPHRPs, suggests a trade‐off between a strategic and operational focus for HR professionals, and also indicates the value of embedding HR practice execution in operational administration. © 2013 Wiley Periodicals, Inc.
HIV‐1 Reverse transcriptase is capable of elongating derivatives of sequence specific noncomplementary oligodeoxynucleotides
Authors: Martyanov Igor V. , Zakharova Ol'Ga D. , Sottofattori Enzo , Maksakova Galiya A. , Pyshnyi Dmitry V. , Mazzei Mauro , Balbi Alessandro , Litvak Simon , Tarrago‐Litvak Laura , Nevinsky Georgy A.
Published: November 2013

We have carried out a comparison of KM and Vmax values for various primers in the polymerization reaction catalyzed by the HIV‐1 RT. The affinity of RT for complementary d(pT)6 containing two different 5′‐end pyranone derivatives was 2–3 orders of magnitude higher (KM = 3–15 nM) than that of d(pT)6 (KM = 12.6 mM). Oligodeoxynucleotides (ODNs) noncomplementary to poly(A) template were not elongated by RT. However, derivatives of d(CAGGTG) containing the 5′‐terminal chromone and coumarin related groups were efficient primers showing KM (30–300 nM) and Vmax (75–93%) values comparable with that for d(pT)10 (800 nM; 100%). The [d(CAGGTG)]ddT ODN derivatives were effective inhibitors of RT. The primer function of derivatives of noncomplementary ODNs appears to be due to the additional interactions of their 5′‐terminal groups with the enzyme tRNA‐binding site.
Prooxidant character of flavonoid cytotoxicity: Structure‐activity relationships
Authors: Dičkancaitė Eglè , Nemeikaitė Aušra , Kalvelytė Audronė , Čėnas Narimantas
Published: November 2013

The action of flavonoids on bovine leukemia virus‐transformed lamb fibroblasts (line FLK) and HL‐60 cells was accompanied by lipid peroxidation, their toxicity was partly prevented by iron chelator desferrioxamine and antioxidant N,N′‐diphenyl‐p‐phenylene diamine. This pointed out to the involvement of oxidative stress in flavonoid cytotoxicity. The concentration of compound for 50% survival of FLK cells (cL50) did not show correlation with Polarographic oxidation half‐peak potential (Ep/2) and/or partition coefficient (log P) of flavonoids; however, their toxicity to HL‐60 cells was described by equation log cL50 (μM) = 3.0161 + 1.1099 Ep/2 (V) — 0.3369 log P. The toxicity of quercetin was partly prevented by nontoxic concentrations of other flavonoids examined, thus pointing out to potential neutralization of quercetin cytotoxicity by intake of flavonoid mixtures.
Nucleotide sequencing of S‐RNA segment and sequence analysis of the nucleocapsid protein gene of the newly isolated akabane virus PT‐17 strain
Authors: Chang Chia‐Wei , Liao Yung‐Kung , Su Vincent , Farh Lynn , Shiuan David
Published: November 2013

The nucleotide sequences of the S‐RNA of Akabane viruses JaGAr‐39, OBE‐1, Iriki and the newly isolated PT‐17 strains and the Aino virus were determined and compared. The results reveal that the S‐RNAs of the four Akabane strains share 96.9% homology in nucleotide sequences. Only one amino acid difference out of the 233 amino acids of the nucleocapsid protein (N) and three amino acid differences in the 91 amino acids of the nonstructural protein (NSs) were found among the Akabane viruses. Amino acid sequences of N and NSs proteins of the Aino virus have approximately 80% identity as compared with the Akabane viruses. The results also demonstrate that the four Akabane viruses and the Aino virus can be clearly differentiated by RFLP (restriction fragments length polymorphism) analysis using RT‐PCR generated nucleocapsid protein genes and digested with HaeIII and HindIII. The phylogenetic tree based on the UPGMA (Unweighted Pair Group Method with Arithmetic Mean) analysis of the sequences of nucleocapsid protein genes and the S‐DNAs revealed that the newly isolated PT‐17 strain is most closely related to Iriki strain, than the JaGAr‐39 or OBE‐1 strains.
Cytochrome C oxidase from eucaryotes but not from procaryotes is allosterically inhibited by ATP
Authors: Follmann Kara , Arnold Susanne , Ferguson‐Miller Shelagh , Kadenbach Bernhard
Published: November 2013

The activity of reconstituted cytochrome c oxidase from bovine heart but not from Rhodobacter sphaeroides is allosterically inhibited by intraliposomal ATP, which binds to subunit IV. The activity of cytochrome c oxidase of wild‐type yeast and of a subunit VIa‐deleted yeast mutant, measured with Tween 20‐solubilized mitochondria in the presence of an ATP‐regenerating system, was also allosterically inhibited by ATP, indicating the general validity of this mechanism of “respiratory control” in eucaryotic cytochrome c oxidases (Arnold and Kadenbach, Eur. J. Biochem. (1997) 249, 350–354). Deletion of subunit Via changes the biphysic into monophysic kinetics of the yeast enzyme in the presence of ADP. A tenfold higher amount of horse heart cytochrome c, as compared to yeast cytochrome c, was required to relieve the ATP inhibition of the yeast enzyme.
Acute‐phase induced phosphorylation of rat liver nucleoprotein P70 modulates its binding affinity for the haptoglobin gene
Authors: Grigorov Ilijana , Milosavljević Tanja , Petrović Miodrag
Published: November 2013

An increase in the binding affinity of rat liver trans‐acting nucleoprotein p70 for the hormone responsive element of the rat haptoglobin gene in acute‐phase reactions has implicated a posttranslational modification. This investigation examines the proposed acute‐phase related structural alterations of p70 using an in vitro phosphorylation/dephosphorylation assay and selective digestion of p70 with Staphylococcal aureus V8 protease. The results show that p70 requires phosphorylation to express its DNA‐binding ability. Selective proteolysis of p70 provided evidence that acute‐phase induced phosphorylation of this protein alters its conformation in such a way that its DNA‐binding ability is increased.
Rapid generation of balanced trial distributions for discrimination learning procedures: A technical note
Authors: Gerard Christophe J. , Mackay Harry A. , Thompson Brooks , McIlvane William J.
Published: November 2013

We describe novel computer algorithms for rapid, sometimes virtually instantaneous generation of trial sequences needed to instrument many behavioral research procedures. Implemented on typical desktop or laptop computers, the algorithms impose constraints to forestall development of undesired stimulus control by position, recent trial outcomes, and other variables that could impede simple and conditional discrimination learning. They yield trial‐by‐trial lists of sequences that can serve (1) as inputs to procedure control software or (2) in generating templates for constructing sessions for implementation by hand or machine.
A New Iodonium Ylide‐Based Three‐Component Reaction Leading to 2‐Spirosubstituted Dihydrofurans under Microwave Irradiation
Authors: Tu Xing‐Chao , Yu Yan , Tu Man‐Su , Jiang Bo , Li Chao , Tu Shu‐Jiang
Published: November 2013

A new iodonium ylide‐based three‐component reaction for the synthesis of highly functionalized 2‐spirosubstituted dihydrofurans starting from readily available common reactants has been developed under microwave irradiation. The procedure is facile, avoiding time‐consuming and costly syntheses, tedious work‐up and purifications of precursors as well as protection/deprotection of functional groups. This method is very efficient because of short reaction times and easy work‐up and provides an efficient strategy for the construction of the polysubstituted spiro dihydrofuran skeleton.
Tandem Aza‐Michael–Aldol Reactions: One‐Pot Synthesis of Functionalized Piperidine Derivatives
Authors: Huang Fu‐Ping , Dong Zhen‐Wen , Zhang Hai‐Rui , Hui Xin‐Ping
Published: November 2013

A series of novel functionalized piperidine derivatives were conveniently prepared by three‐component tandem aza‐Michael–Aldol reactions of α,β‐unsaturated enones with substituted sulfonamides in the presence of DBU. The structures of the compounds synthesized were characterized by spectral data and X‐ray crystal diffraction.
Synthesis and Herbicidal Activity of 2‐Aroxy‐propanamides Containing Pyrimidine and 1,3,4‐Thiadiazole Rings
Authors: Liu Man‐Yun , Shi De‐Qing
Published: November 2013

A series of novelN‐{5‐[2‐(4,6‐dimethoxy‐pyrimidin‐2‐yloxy)‐phenyl]‐[1,3,4]thiadiazol‐2‐yl}2‐aroxy‐propanamides were designed and synthesized by the multistep reactions. 2‐(4,6‐Dimethoxy‐pyrimidin‐2‐yloxy)‐benzaldehyde (1) reacted with aminothiourea to yield2, which undergoes ring closure to give 5‐[2‐(4,6‐dimethoxy‐pyrimidin‐2‐yloxy)‐phenyl]‐[1,3,4]thiadiazol‐2‐amine (3) in the presence of ferric chloride in refluxing ethanol.3reacted with 2‐aroxy‐propionyl chlorides to give the target compounds4a,4b,4c,4d,4e,4f,4g,4h,4iin moderate to good yields. Their structures were confirmed by IR,1H‐NMR, EIMS, and elemental analyses. The preliminary bioassay indicated that some of them displayed moderate to good selective herbicidal activity againstBrassica campestris L. at the concentration of 100 mg/L. However, these compounds did not possess inhibitory activity againstEchinochloa crus‐galliat the tested concentrations.
Interaction investigations of HipA binding to HipB dimer and HipB dimer + DNA complex: a molecular dynamics simulation study
Authors: Li Chaoqun , Wang Yaru , Wang Yan , Chen Guangju
Published: November 2013

We carried out molecular dynamics simulations and free energy calculations for a series of ternary and diplex models for the HipA protein, HipB dimer, and DNA molecule to address the mechanism of HipA sequestration and the binding order of events from apo HipB/HipA to 2HipA + HipB dimer + DNA complex. The results revealed that the combination of DNA with the HipB dimer is energetically favorable for the combination of HipB dimer with HipA protein. The binding of DNA to HipB dimer induces a long‐range allosteric communication from the HipB2‐DNA interface to the HipA–HipB2 interface, which involves the closeness of α1 helices of HipB dimer to HipA protein and formations of extra hydrogen bonds in the HipA–HipB2 interface through the extension of α2/3 helices in the HipB dimer. These simulated results suggested that the DNA molecule, as a regulative media, modulates the HipB dimer conformation, consequently increasing the interactions of HipB dimer with the HipA proteins, which explains the mechanism of HipA sequestration reported by the previous experiment. Simultaneously, these simulations also explored that the thermodynamic binding order in a simulated physiological environment, that is, the HipB dimer first bind to DNA to form HipB dimer + DNA complex, then capturing strongly the HipA proteins to form a ternary complex, 2HipA + HipB dimer + DNA, for sequestrating HipA in the nucleoid. Copyright © 2013 John Wiley & Sons, Ltd.
Combination of fluorescence microscopy and nanomotion detection to characterize bacteria
Authors: Aghayee S. , Benadiba C. , Notz J. , Kasas S. , Dietler G. , Longo G.
Published: November 2013

Antibiotic‐resistant pathogens are a major health concern in everyday clinical practice. Because their detection by conventional microbial techniques requires minimally 24 h, some of us have recently introduced a nanomechanical sensor, which can reveal motion at the nanoscale. By monitoring the fluctuations of the sensor, this technique can evidence the presence of bacteria and their susceptibility to antibiotics in less than 1 h. Their amplitude correlates to the metabolism of the bacteria and is a powerful tool to characterize these microorganisms at low densities. This technique is new and calls for an effort to optimize its protocol and determine its limits. Indeed, many questions remain unanswered, such as the detection limits or the correlation between the bacterial distribution on the sensor and the detection's output. In this work, we couple fluorescence microscopy to the nanomotion investigation to determine the optimal experimental protocols and to highlight the effect of the different bacterial distributions on the sensor. Copyright © 2013 John Wiley & Sons, Ltd.
Insulin receptor‐insulin interaction kinetics using multiplex surface plasmon resonance
Authors: Subramanian Kannan , Fee Conan J. , Fredericks Rayleen , Stubbs Richard S. , Hayes Mark T.
Published: November 2013

Type 2 diabetes affects millions of people worldwide, and measuring the kinetics of insulin receptor‐insulin interactions is critical to improving our understanding of this disease. In this paper, we describe, for the first time, a rapid, real‐time, multiplex surface plasmon resonance (SPR) assay for studying the interaction between insulin and the insulin receptor ectodomain, isoform A (eIR‐A). We used a scaffold approach in which anti‐insulin receptor monoclonal antibody 83–7 (Abcam, Cambridge, UK) was first immobilized on the SPR sensorchip by amine coupling, followed by eIR‐A capture. The multiplex SPR system (ProteOn XPR36™, Bio‐Rad Laboratories, Hercules, CA) enabled measurement of replicate interactions with a single, parallel set of analyte injections, whereas repeated regeneration of the scaffold between measurements caused variable loss of antibody activity. Interactions between recombinant human insulin followed a two‐site binding pattern, consistent with the literature, with a high‐affinity site (dissociation constant KD1 = 38.1 ± 0.9 nM) and a low‐affinity site (KD2 = 166.3 ± 7.3 nM). The predominantly monomeric insulin analogue Lispro had corresponding dissociation constants KD1 = 73.2 ± 1.8 nM and KD2 = 148.9 ± 6.1 nM, but the fit to kinetic data was improved when we included a conformational change factor in which the high‐affinity site was converted to the low‐affinity site. The new SPR assay enables insulin‐eIR‐A interactions to be followed in real time and could potentially be extended to study the effects of humoral factors on the interaction, without the need for insulin labeling. Copyright © 2013 John Wiley & Sons, Ltd.
Conformational dynamics of individual antibodies using computational docking and AFM
Authors: Chaves Rui C. , Teulon Jean‐Marie , Odorico Michael , Parot Pierre , Chen Shu‐wen W. , Pellequer Jean‐Luc
Published: November 2013

Molecular recognition between a receptor and a ligand requires a certain level of flexibility in macromolecules. In this study, we aimed at analyzing the conformational variability of receptors portrayed by monoclonal antibodies that have been individually imaged using atomic force microscopy (AFM). Individual antibodies were chemically coupled to activated mica surface, and they have been imaged using AFM in ambient conditions. The resulting topographical surface of antibodies was used to assemble the three subunits constituting antibodies: two antigen‐binding fragments and one crystallizable fragment using a surface‐constrained computational docking approach. Reconstructed structures based on 10 individual topographical surfaces of antibodies are presented for which separation and relative orientation of the subunits were measured. When compared with three X‐ray structures of antibodies present in the protein data bank database, results indicate that several arrangements of the reconstructed subunits are comparable with those of known structures. Nevertheless, no reconstructed structure superimposes adequately to any particular X‐ray structure consequence of the antibody flexibility. We conclude that high‐resolution AFM imaging with appropriate computational reconstruction tools is adapted to study the conformational dynamics of large individual macromolecules deposited on mica. Copyright © 2013 John Wiley & Sons, Ltd.
Retrospective evaluation of dosimetric quality for prostate carcinomas treated with 3D conformal, intensity modulated and volumetric modulated arc radiotherapy
Authors: Crowe Scott B. , Kairn Tanya , Middlebrook Nigel , Hill Brendan , Christie David R. H. , Knight Richard T. , Kenny John , Langton Christian M. , Trapp Jamie V.
Published: November 2013

Introduction

This study examines and compares the dosimetric quality of radiotherapy treatment plans for prostate carcinoma across a cohort of 163 patients treated across five centres: 83 treated with three‐dimensional conformal radiotherapy (3DCRT), 33 treated with intensity modulated radiotherapy (IMRT) and 47 treated with volumetric modulated arc therapy (VMAT).

Methods

Treatment plan quality was evaluated in terms of target dose homogeneity and organs at risk (OAR), through the use of a set of dose metrics. These included the mean, maximum and minimum doses; the homogeneity and conformity indices for the target volumes; and a selection of dose coverage values that were relevant to each OAR. Statistical significance was evaluated using two‐tailed Welch's T‐tests. The Monte Carlo DICOM ToolKit software was adapted to permit the evaluation of dose metrics from DICOM data exported from a commercial radiotherapy treatment planning system.

Results

The 3DCRT treatment plans offered greater planning target volume dose homogeneity than the other two treatment modalities. The IMRT and VMAT plans offered greater dose reduction in the OAR: with increased compliance with recommended OAR dose constraints, compared to conventional 3DCRT treatments. When compared to each other, IMRT and VMAT did not provide significantly different treatment plan quality for like‐sized tumour volumes.

Conclusions

This study indicates that IMRT and VMAT have provided similar dosimetric quality, which is superior to the dosimetric quality achieved with 3DCRT.
Hepatitis E virus open reading frame 3 protein interacts with porcine liver‐specific plasminogen and α2‐antiplasmin
Authors: Zhou Yan , Geng Yansheng , Yang Jun , Zhao Chenyan , Harrison Tim J. , Wang Youchun
Published: November 2013

Hepatitis E virus (HEV) is a hepatotropic virus that causes acute and, occasionally, chronic viral hepatitis. At least four recognized genotypes of mammalian HEV have been identified. Genotypes 3 and 4 are zoonotic in humans and pigs are the major host. The 7.2 kb genome of HEV contains three open reading frames: ORF1, ORF2, and ORF3. ORF1 encodes a nonstructural protein and the ORF2 protein is the main capsid protein but the precise functions of ORF3 protein remain obscure. To explore the role of ORF3 in the porcine host, the genotype 4 ORF3 protein was used for yeast two‐hybrid screening to find cellular binding partners encoded by a pig liver cDNA library, two porcine liver specific proteins, plasminogen and α2‐antiplasmin were identified. Their interactions with the ORF3 protein were validated by chemiluminescent Co‐Immunoprecipitation assay and by Western blotting in Co‐IP and His pull‐down assay. The biological significance of these interactions and their possible role in the HEV infection are discussed. J. Med. Virol. 86:487–495, 2014. © 2013 Wiley Periodicals, Inc.
Overexpression of Wnt3a facilitates the proliferation and neural differentiation of neural stem cells in vitro and after transplantation into an injured rat retina
Authors: Yang Xi‐Tao , Bi Yong‐Yan , Chen Er‐Tao , Feng Dong‐Fu
Published: November 2013

Neural stem cell‐based therapy is a promising option for repair after injury. However, poor stem cell proliferation and insufficient differentiation of the stem cells into neurons are still difficult problems. The present study investigated whether transplantation of neural stem cells (NSCs) genetically modified to express Wnt3a is a promising approach to overcome these difficulties. We explored the possibility that Wnt3a might contribute to the therapeutic effect of NSC transplantation in retinal repair. The relative promotion of proliferation and neural differentiation by modified NSCs was investigated in a rat model of optic nerve crush. A recombinant lentivirus (Lenti‐Wnt3a) was engineered to express Wnt3a. NSCs infected with control lentivirus (Lenti‐GFP) or Lenti‐Wnt3a were transplanted into the subretinal space immediately after the optic nerve crush. The proliferation and neural differentiation activity of the NSCs were assessed in vitro and in vivo. Overexpression of Wnt3a in NSCs induced activation of Wnt signaling, promoted proliferation, and directed the differentiation of the NSCs into neurons both in vitro and in vivo. Our study suggests that Wnt3a can potentiate the therapeutic benefits of NSC‐based therapy in the injured retina. © 2013 Wiley Periodicals, Inc.
Starbucks Exceptionalism: An Institutional Ethnographic Exploration of Coffee Culture in America
Authors: Gavin Diane
Published: November 2013

Over a 5‐year period, from 2008 to 2013, Starbucks locations across the United States were visited and revisited to understand the growing cultural phenomenon that took hold in American society regarding consumers and Starbucks's coffee products. Written in an enhanced ethnographic style, in which presentational techniques are drawn from creative nonfiction and immersion journalism, the purpose of the current study is to apply a political theory lens in American culture, Seymour Lipset's (1966/1996) theory of American exceptionalism, to an American corporation, Starbucks Coffee. As with any ethnographic study, field work comes from a blend of immersion in the field and secondary data sources. For this study, the researcher visited more than 100 individual Starbucks locations across the United States, spoke with hundreds of individuals in these locations, and conducted a one‐word, quick‐survey response with 200 participants who were criterion selected based on their visiting a Starbucks on the day of being asked the question. The participants were randomly asked on the street as well as through Facebook. Descriptive and thematic coding processes were conducted on the participants' responses. The results indicated that while Starbucks Corporation messaging indicates the presence of Starbucks exceptionalism, fragmentation in consumers' minds may be occurring and Starbucks exceptionalism may not be as strong as it was a few years ago. As Starbucks moves into the Far Asian and Southeast Asian marketplaces, further study is needed to determine whether Starbucks exceptionalism will stay intact or shift to reflect other cultural influences.
How Government Venture Capital Guiding Funds Work in Financing High‐Tech Start‐Ups in China: A ‘Strategic Exchange’ Perspective
Authors: Wang Jinmin , Wang Jing , Ni Hua , He Shaowei
Published: November 2013

Under its specific institutional context, the public/private partnership approach needs to be interpreted differently from strategic level to operational level in China to improve the performance of venture capital guiding funds.
Appropriate gastrectomy resection margins for early gastric carcinoma
Authors: Kim Beom Su , Oh Seong Tae , Yook Jeong Hwan , Kim Hee Sung , Lee In Seob , Kim Byung Sik
Published: November 2013

Objective

In Korea and Japan, early gastric cancer (EGC) accounts for >50% of all gastric cancers. Here, we propose recommendations for the optimal distance from the tumor to the resection margins when evaluating EGC.

Summary of Background Data

There are very few guidelines regarding the distance from the EGC tumor to the resection margins.

Methods

We evaluated 2,081 patients who underwent gastrectomy for EGC between January 1989 and May 2000. We subdivided tumors according to the distance from the proximal margin: ≤1, >1, ≤10, >10, ≤30, or >30 mm.

Results

Three of five patients demonstrating distances ≤1 mm between the tumor and gross proximal margin were microscopically positive. No patients with gross proximal margins >1, ≤10, >10, or ≤30 mm were microscopically positive. There were no statistical differences in rates of microscopically positive margin, reresection, or reoperation between groups (P > 0.05). In addition, there were statistical differences in terms of tumor recurrence and disease‐related death between groups (P > 0.05).

Conclusions

When the resection margins are clear, we propose that margins >1 mm are adequate for EGC gastrectomy. J. Surg. Oncol. 2014 109:198–201. © 2013 Wiley Periodicals, Inc.
Should a loop ileostomy closure in rectal cancer patients be done during or after adjuvant chemotherapy?
Authors: Tulchinsky Hagit , Shacham‐Shmueli Einat , Klausner Joseph M. , Inbar Moshe , Geva Ravit
Published: November 2013

Background and Objective

The influence of ileostomy closure timing on surgical and oncologic outcome was investigated in patients with locally advanced rectal cancer receiving adjuvant chemotherapy after low anterior resection.

Methods

Consecutive patients diagnosed with T3‐4/N+ rectal cancer, treated by neoadjuvant chemoradiation and low anterior resection during 2000–2012 were retrospectively evaluated. Patients undergoing closure during adjuvant chemotherapy (Group A) were compared to patients undergoing closure after completing chemotherapy (Group B).

Results

A total of the 165 patients met inclusion criteria, of whom 104 received adjuvant chemotherapy (25 in Group A and 79 in Group B). The pathologic stage was higher in Group B (P = 0.015). The rates of postoperative complications were similar (16% for Group A and 15% for Group B, P = 0.88), as was hospital stay (mean 5.78 days for Group A and 6.25 days for Group B, P = 0.7). There was no significant difference in recurrence rate and overall survival between the groups.

Conclusions

Referral to ileostomy closure in relation to adjuvant chemotherapy is influenced by pathologic stage. Early referral appears to be reserved to a small number of patients with lower pathologic stage. Timing of ileostomy closure does not change short‐ or long‐term results. J. Surg. Oncol. 2014 109:266–269. © 2013 Wiley Periodicals, Inc.
Expression of apoptosis regulating proteins identifies stage II and III colon cancer patients with high risk of recurrence
Authors: Belt Eric J.Th. , Stockmann Hein B.A.C. , Delis‐van Diemen Pien M. , Bril Herman , Tijssen Marianne , van Essen Hendrik F. , Heymans Martijn W. , Beliën Jeroen A.M. , Carvalho Beatriz , Cillessen Saskia A.G.M. , Meijer Gerrit A.
Published: November 2013

Background and Objectives

Deregulation of apoptosis related genes may be associated with poor outcome in cancer. Aim of the present study was to investigate the prognostic role of expression levels of apoptosis related proteins in stage II and III colon cancer.

Methods

From tumor samples of 386 stage II and III colon cancer patients, DNA was isolated and tissue microarrays were constructed. Expression of Bcl‐2, Bcl‐X, BAX, XIAP, Fas, FasL and c‐FLIP was evaluated and PCR‐based microsatellite instability analysis was performed.

Results

High FasL expressing tumors were associated with high disease recurrence rates in stage II colon cancer patients overall, as was low Bcl‐X expression in microsatellite stable stage II patients. In stage II patients, a multivariable model based on FasL and Bcl‐XL expression revealed a significant association with disease free survival (DFS). In stage III colon cancer patients, low Bcl‐2, low BAX and low Fas expression levels were associated with worse outcome. In these patients a multivariable model based on angioinvasion and Bcl‐2, Fas and FasL expression was significantly associated with DFS.

Conclusions

Stage II patients with low Bcl‐X and high FasL protein expression levels and stage III patients with low Fas, high FasL and low Bcl‐2 expression could be considered as high risk for disease recurrence. J. Surg. Oncol. 2014 109:255–265. © 2013 Wiley Periodicals, Inc.
Fluorescently labeled chimeric anti‐CEA antibody improves detection and resection of human colon cancer in a patient‐derived orthotopic xenograft (PDOX) nude mouse model
Authors: Metildi Cristina A. , Kaushal Sharmeela , Luiken George A. , Talamini Mark A. , Hoffman Robert M. , Bouvet Michael
Published: November 2013

Background and Objectives

The aim of this study was to evaluate a new fluorescently labeled chimeric anti‐CEA antibody for improved detection and resection of colon cancer.

Methods

Frozen tumor and normal human tissue samples were stained with chimeric and mouse antibody‐fluorophore conjugates for comparison. Mice with patient‐derived orthotopic xenografts (PDOX) of colon cancer underwent fluorescence‐guided surgery (FGS) or bright‐light surgery (BLS) 24 hr after tail vein injection of fluorophore‐conjugated chimeric anti‐CEA antibody. Resection completeness was assessed using postoperative images. Mice were followed for 6 months for recurrence.

Results

The fluorophore conjugation efficiency (dye/mole ratio) improved from 3–4 to >5.5 with the chimeric CEA antibody compared to mouse anti‐CEA antibody. CEA‐expressing tumors labeled with chimeric CEA antibody provided a brighter fluorescence signal on frozen human tumor tissues (P = 0.046) and demonstrated consistently lower fluorescence signals in normal human tissues compared to mouse antibody. Chimeric CEA antibody accurately labeled PDOX colon cancer in nude mice, enabling improved detection of tumor margins for more effective FGS. The R0 resection rate increased from 86% to 96% with FGS compared to BLS.

Conclusion

Improved conjugating efficiency and labeling with chimeric fluorophore‐conjugated antibody resulted in better detection and resection of human colon cancer in an orthotopic mouse model. J. Surg. Oncol. 2014 109:451–458. © 2013 Wiley Periodicals, Inc.
Prognostic value of kisspeptin expression in nasopharyngeal carcinoma
Authors: Yuan Tai‐Ze , Zhang Huan‐Huan , Tang Qiong‐Fen , Zhang Qiong , Li Jian , Liang Yin , Huang Lai‐Ji , Zheng Rong‐Hui , Deng Jin , Zhang Xiu‐Ping
Published: November 2013

Objectives/Hypothesis

The KiSS‐1 gene has been reported to serve as a metastasis suppressor gene in various human malignancies. However, no information is available regarding the role of the KiSS‐1 gene or its gene product kisspeptin in nasopharyngeal carcinoma.

Study Design

Retrospective study.

Methods

Kisspeptin and its receptor AXOR12 expression were assessed using immunohistochemistry in paraffin‐embedded tumor tissues from 140 patients diagnosed with nasopharyngeal carcinoma. Immunoreactivity was quantified, and its relationships with patients' clinical parameters and survival were analyzed.

Results

Using a 50% cutoff level, the immunoreactivities of kisspeptin and AXOR12 were divided into low and high expression. The expression levels of kisspeptin and AXOR12 in nasopharyngeal carcinoma were well correlated with each other (rs = 19.31, P < 0.01). Low expression of kisspeptin in nasopharyngeal carcinoma was correlated with clinical stage (P = 0.01), N stage (P = 0.03), and metastasis (P = 0.02). Patients with low kisspeptin expression had poorer distant metastasis‐free survival than those with high kisspeptin expression (75.32% vs. 83.79%, P = 0.02). Although neither kisspeptin nor AXOR12 were found to be prognostic factors for overall survival, kisspeptin was determined to be an independent prognostic factor for distant metastasis‐free survival (P = 0.03) using multivariate analysis.

Conclusion

In this study, we report for the first time that low kisspeptin expression in nasopharyngeal carcinoma is correlated with poor clinical outcome; kisspeptin could serve as an independent prognostic marker for metastasis in nasopharyngeal carcinoma.

Level of Evidence

N/A. Laryngoscope, 124:E167–E174, 2014
Using Static and Dynamic Indicators to Evaluate Soil Physical Quality in a Sicilian Area
Authors: Iovino Massimo , Castellini Mirko , Bagarello Vincenzo , Giordano Giuseppe
Published: November 2013

Both capacitive indicators derived from the water retention curve and dynamic measurements of the flow‐weighted mean pore radius, R0, were used to assess the soil physical quality of two agricultural areas (cropland and olive orchard) and two natural areas (grassland and managed woodlot plantation) potentially subject to soil degradation. The overall idea of the study was to investigate whether a dynamic indicator quantitatively derived from hydraulic conductivity measurements could be used to supplement the traditionally applied capacitive indicators retrieved from water retention measurements. According to the available criteria, only the surface layer of the cropland site showed optimal soil physical quality. In the grassland and woodlot sites, the physical quality was deteriorated also as a consequence of compaction because of grazing. Overall, the physical quality was better in tilled than nontilled soils. The optimal soil in terms of capacitive indicators had hydraulic conductivity close to saturation that was intermediate among the different land uses, and it remained 1·3–1·9 times higher than that observed in the natural sites even when the largest pores emptied. A depth effect on R0 was observed only when larger macropores were activated. It was suggested that water transmission parameters are more affected by changes in large pore domain. The plant available water content and Dexter's S‐index showed inverse statistically significant regressions with R0. The empirical relationships were physically convincing given that, at increasing R0, the contribution of macropores increases, water is transmitted faster below the root zone and the soil's ability to store water is reduced. Copyright © 2013 John Wiley & Sons, Ltd.