bluuebunny/crossref_metadata_2025_split huggingface dataset:
The electrophoretic mobility of DNA fragments differing by a single 3′‐terminal nucleotide in an automated capillary DNA sequencer
Authors: Nguyen Trung V. , Murray Vincent 
Published: August 2012

ABSTRACTThe electrophoretic mobility of DNA fragments that differ by a single 3′‐terminal nucleotide was assessed by capillary electrophoresis. This was accomplished using dideoxy sequencing with a 5′‐fluorescently labelled primer to generate DNA fragments with 3′‐hydrogen ends. The resulting DNA fragments were electrophoresed on the ABI 3730 automated capillary sequencer, and the data were analysed with the GeneMapper software to determine the electrophoretic mobility differences on addition of a 3′‐terminal nucleotide. It was found that the 3′‐terminal nucleotide gave rise to different electrophoretic mobility profiles depending on the identity of the terminal nucleotide. The apparent electrophoretic mobility was (faster) –C > −A > −T > −G (slower). The C‐terminated fragments were the fastest and the G‐terminated fragments the slowest, relative to other nucleotides. It was proposed that the terminal nucleotide effect was due to changes in partial net charges on the nucleotides that resulted in alterations in the electrophoretic mobility of the DNA fragments in the automated capillary DNA sequencer. Other alternative explanations are also discussed. Copyright © 2012 John Wiley & Sons, Ltd.
Quantitation and characterization of process impurities and extractables in protein‐containing solutions using proton NMR as a general tool
Authors: Skidmore Ken , Hewitt Daniel , Kao Yung‐Hsiang 
Published: August 2012

AbstractThe ability to detect and quantitate a variety of components in solution has become increasingly important in carrying out efficient and rigorous validation studies for biopharmaceutical manufacturing processes. Here, we demonstrate the general applicability of NMR spectroscopy for the identification and quantitation of leachables and other impurities in protein‐based drugs, at low levels previously unattainable in protein‐containing solutions. With improved NMR technology (i.e., CryoProbes) and the application of a Carr‐Purcell‐Meiboom‐Gill pulse sequence (CPMG) to attenuate protein signals, we have been able to use NMR to quantify impurities in a protein‐based biopharmaceutical product at ∼1 μg mL−1. The data indicate that NMR spectra can be used to quantitate a range of impurities, from small molecule components to higher molecular weight leachables, without removing protein from solution. Furthermore, quantitation of impurities by NMR is reliable and accurate enough for biopharmaceutical process validation, even for high molecular weight extractables whose structures are not precisely known. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012
Cloning and expression of a functional core streptavidin in Pichia pastoris: Strategies to increase yield
Authors: Casteluber Marisa C. F. , Damasceno Leonardo M. , da Silveira Wendel B. , Diniz Raphael H. S. , Passos Frederico J. V. , Passos Flávia M. L. 
Published: August 2012

AbstractStreptavidin is widely used as an analytical tool and affinity tag together with biotinylated surfaces or molecules. We report for the first time a simple strategy that yields high biomass of a Pichia pastoris strain containing a methanol induced core streptavidin (cStp) gene. Three factors were evaluated for biomass production: glycerol concentration, aeration, and feed flow rates in a bioreactor. Recycling of recombinant cells, either free or immobilized, was investigated during induction. Concentration of 2.0 M glycerol, feeding flow rate of 0.11 mL min−1, and aeration by air injection dispersed with a porous stone combined with agitation at 500 rpm were the set of conditions resulting into maximum biomass yield (150 g L−1). These parameters yielded 4.0 g L−1 of cStp, after 96 h of induction. Recombinant biomass was recycled twice before being discarded, which can reduce production costs and simplify the process. Immobilized P. pastoris biomass produced 2.94 and 1.70 g L−1 of cStp in the first and second induction cycle, respectively. Immobilization and recycling of recombinant P. pastoris biomass opens new possibilities as a potential strategy to improve volumetric productivity for heterologous protein expression. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012
Use of enzymes in extraction of polyhydroxyalkanoates produced by Cupriavidus necator
Authors: Neves Andréia , Müller José 
Published: August 2012

AbstractPoly(3‐hydroxybutyrate) (P(3HB)) and its copolymer poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate) (P(3HB‐co‐3HV), are biodegradable thermoplastic polymers. They are members of the polyhydroxyalkanoate (PHA) family, synthesized and accumulated as a carbon and energy reserve by a variety of microorganisms. The aim of this study was to evaluate the use of the proteases Corolase® L10, Alcalase® 2.4L, Corolase® 7089 and Protemax® FC and glycosidases Celumax® BC, Rohament® CL and Rohalase® Barley for the recovery of P(3HB) and P(3HB‐co‐3HV) synthesized by Cupriavidus necator. The enzyme Celumax® BC provided better lysis of the bacterial cell membrane and the results for the optimization of the operating conditions showed that this enzyme is most stable in acetate buffer at pH 4.0, bath at 60°C, hydrolysis time of 1 h and concentration of 0.02% (w/w). The optimization of the operating conditions showed that the enzyme Celumax® BC provided better lysis of the bacterial cell in acetate buffer at pH 4.0, bath at 60°C, hydrolysis time of 1 h and concentration of 0.02% (w/w). These conditions resulted in lysis of the membrane of the bacteria with a recovery of 93.2% P(3HB‐co‐3HV) with 94% purity. The results showed that the use of enzymes for the polymer extraction is an efficient process that assists in the cell disruption of Cupriavidus necator. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012
Type‐I Dyotropic Reactions: Understanding Trends in Barriers
Authors: Fernández Israel , Bickelhaupt F. Matthias , Cossío Fernando P. 
Published: August 2012

AbstractTo understand the factors that control the activation barrier of type‐I 1,2‐dyotropic reactions (X‐EH2‐CH2‐X*→X*‐EH2‐CH2‐X, with E=C and Si, X=H, CH3, SiH3, F to I) and trends therein as a function of the migrating groups X, we have explored ten archetypal model reactions of this class using relativistic density functional theory (DFT) at ZORA‐OLYP/TZ2P. The main trends in reactivity are rationalized using the activation strain model of chemical reactivity, which had to be extended from bimolecular to unimolecular reactions. Thus, the above type‐I dyotropic reactions can be conceived as a relative rotation of the CH2CH2 and [X⋅⋅⋅X] fragments in X‐CH2‐CH2‐X. The picture that emerges from these analyses is that reduced CX bonding in the transition state is the origin of the reaction barrier. Also the trends in reactivity on variation of X can be understood in terms of how sensitive the CX interaction is towards adopting the transition‐state geometry. A valence bond analysis complements the analyses and confirms the picture emerging from the activation strain model.
β‐Cyclodextrin Duplexes That Are Connected through Two Disulfide Bonds: Potent Hosts for the Complexation of Organic Molecules
Authors: Grishina Anastasia , Stanchev Stancho , Kumprecht Lukáš , Buděšínský Miloš , Pojarová Michaela , Dušek Michal , Rumlová Michaela , Křížová Ivana , Rulíšek Lubomír , Kraus Tomáš 
Published: August 2012

AbstractNew tubular host molecules, which are composed of two β‐cyclodextrin macrocycles that are connected through two disulfide bonds, have been prepared by the air‐promoted oxidation of 6I,6IV‐dideoxy‐6I,6IV‐disulfanyl‐β‐cyclodextrin in aqueous solution. This reaction leads to three products: monomeric intramolecular disulfide and two dimeric species, which are termed as “non‐eclipsed” and “eclipsed” cyclodextrin duplexes. Oxidation at a concentration of the starting thiol of 0.1 mM gave the intramolecular disulfide as the major product whereas a concentration in the millimolar range afforded the dimeric species as the dominant products. The tubular structure of the “non‐eclipsed” isomer was unequivocally determined by X‐ray analysis. The binding affinities of the duplexes to a wide range of compounds, including fluorescent dyes and clinically used drugs Imatinib and Esomeprazol, were studied in water by ITC. For most guest compounds, the experimentally determined Ka values were in the range 107–108 M−1. These binding affinities are significantly higher than those found in the literature for analogous complexes with native cyclodextrins. In cases of binding of neutral or anionic guest molecules cyclodextrin duplexes outperformed cucurbiturils. A complex between a duplex and Nile blue was used to investigate its ability to penetrate the cytoplasmic membrane of HeLa cells. We found that the complex accumulated in the cell membrane but did not pass into cytosol. Importantly, the complex did not decompose to a significant extent under high dilution in the cellular environment.
Vapochromic Ionic Liquids from Metal–Chelate Complexes Exhibiting Reversible Changes in Color, Thermal, and Magnetic Properties
Authors: Funasako Yusuke , Mochida Tomoyuki , Takahashi Kazuyuki , Sakurai Toshihiro , Ohta Hitoshi 
Published: August 2012

AbstractVapor‐ and gas‐responsive ionic liquids (ILs) comprised of cationic metal‐chelate complexes and bis(trifluoromethanesulfonyl)imide (Tf2N) have been prepared, namely, [Cu(acac)(BuMe3en)][Tf2N] (1 a), [Cu(Bu‐acac)(BuMe3en)][Tf2N] (1 b), [Cu(C12‐acac)(Me4en)][Tf2N] (1 c), [Cu(acac)(Me4en)][Tf2N] (1 d), and [Ni(acac)(BuMe3en)][Tf2N] (2 a) (acac=acetylacetonate, Bu‐acac=3‐butyl‐2,4‐pentanedionate, C12‐acac=3‐dodecyl‐2,4‐pentanedionate, BuMe3en=N‐butyl‐N,N′,N′‐tetramethylethylenediamine, and Me4en=N,N,N′,N′‐trimethylethylenediamine). These ILs exhibited reversible changes in color, thermal properties, and magnetic properties in response to organic vapors and gases. The CuII‐containing ILs are purple and turn blue‐purple to green when exposed to organic vapors, such as acetonitrile, methanol, and DMSO, or ammonia gas. The color change is based on the coordination of the vapor molecules to the cation, and the resultant colors depend on the coordination strength (donor number, DN) of the vapor molecules. The vapor absorption caused changes in the melting points and viscosities, leading to alteration in the phase behaviors. The IL with a long alkyl chain (1 d) transitioned from a purple solid to a brown liquid at its melting point. The NiII‐containing IL (2 a) is a dark red diamagnetic liquid, which turned into a green paramagnetic liquid by absorbing vapors with high DN. Based on the equilibrium shift from four‐ to six‐coordinated species, the liquid exhibited thermochromism and temperature‐dependent magnetic susceptibility after absorbing methanol.
Electrochemical Design of Mesoporous Pt–Ru Alloy Films with Various Compositions toward Superior Electrocatalytic Performance
Authors: Wang Hongjing , Imura Masataka , Nemoto Yoshihiro , Wang Liang , Jeong Hu Young , Yokoshima Tokihiko , Terasaki Osamu , Yamauchi Yusuke 
Published: August 2012

AbstractMesoporous Pt–Ru alloy films with various compositions were synthesized by electrochemical plating in an aqueous surfactant solution. After the removal of surfactants, continuous mesoporous Pt–Ru alloy films possessing uniform mesopores with diameter about 7 nm were obtained. The Ru content in the films could be controlled from 0 to 13 at % by changing the precursor compositions. For all the films, the mesostructural periodicities and the mesopore sizes in the films were not changed. Due to the mesoporous structure and the doped Ru content, our mesoporous Pt–Ru films showed superior electrocatalytic activity for methanol oxidation reaction in comparison with the commercially available Pt catalyst.
Toll‐like receptor‐4 signaling in mantle cell lymphoma
Authors: Wang Lijuan , Zhao Yi , Qian Jianfei , Sun Luhong , Lu Yong , Li Haiyan , Li Yi , Yang Jing , Cai Zhen , Yi Qing 
Published: August 2012

AbstractBACKGROUND:Mantle cell lymphoma (MCL) is an incurable B‐cell malignancy, and patients with this disease have the poorest prognosis among all patients with B‐cell lymphomas. The signaling pathways that trigger MCL escape from immune surveillance are unclear. Because Toll‐like receptors (TLRs) initiate innate and adaptive immune responses against invading pathogens, the authors investigated the impact of TLR signaling in MCL cells.METHODS:TLR expression was examined in MCL cell lines and in primary tumors. The examination focused on TLR4 and its ligand lipopolysaccharide (LPS) on MCL cells and their function on MCL proliferation and immune evasion.RESULTS:MCL cells expressed multiple TLRs, and TLR4 was among the highest expressed molecules. The activation of TLR4 signaling in MCL cells by LPS induced MCL proliferation and up‐regulated the secretion of cytokines like interleukin‐6 (IL‐6), IL‐10, and vascular endothelial growth factor (VEGF). LPS‐pretreated MCL cells inhibited the proliferation and cytolytic activity of T cells by secreted IL‐10 and VEGF, and neutralizing antibodies against these cytokines restored their functions. Similar results were observed in TLR4‐positive/myeloid differentiation 88 (MyD88)‐positive primary lymphoma cells but not in TLR4‐positive/MyD88‐negative primary lymphoma cells from patients with MCL. Knockdown of TLR4 on MCL cells abrogated the effect of LPS on MCL cells in term of cell growth or secretion of the cytokines and evasion of the immune system.CONCLUSIONS:The current results indicated that TLR4 signaling triggers a cascade that leads to MCL growth and evasion from immune surveillance. Thus, TLR4 signaling molecules may be novel therapeutic targets in patients with MCL. Cancer 2013. © 2012 American Cancer Society.
Subjective binge eating with compensatory behaviors: A variant presentation of bulimia nervosa
Authors: Watson Hunna J. , Fursland Anthea , Bulik Cynthia M. , Nathan Paula 
Published: August 2012

AbstractObjectives:To determine whether a variant bulimic‐type presentation, whereby one meets criteria for bulimia nervosa (BN) except that binge eating episodes are not objectively large (i.e., “subjective bulimia nervosa,” SBN), has comparable clinical severity to established eating disorders, particularly BN.Method:Treatment‐seeking adults with BN (N = 112), SBN (N = 28), anorexia nervosa restricting type (AN‐R) (N = 45), and AN‐binge/purge type (AN‐B/P) (N = 24) were compared.Results:Overall, SBN could not be meaningfully distinguished from BN. SBN and BN had equivalent eating pathology, depression and anxiety symptoms, low quality of life, impulsivity, Axis I comorbidity, and lifetime psychiatric history, and comparable clinical severity to AN‐R and AN‐B/P.Discussion:Individuals with SBN, differing from BN only by the smaller size of their binge eating episodes, had a form of eating disorder comparable in clinical severity to threshold AN and BN and warranting clinical attention. Health professionals and the community require greater awareness of this variant to optimize detection, treatment‐seeking, and outcomes. © 2012 by Wiley Periodicals, Inc. (Int J Eat Disord 2013)
Improved bi‐mesa structures optimized for resonator mounting
Authors: Goka Shigeyoshi , Mase Yusuke , Sekimoto Hitoshi 
Published: August 2012

AbstractAn improved bi‐mesa structure optimized for resonator mounting is proposed. To evaluate mounting influences, the frequency differences between clamped and free X‐edge conditions are calculated using a two‐dimensional finite element method in the X–Y′ region. When the outer mesa height was lower than 20% of the thick area, the frequency differences are less than 10% of standard‐type bi‐mesa resonator values. These results indicate that the improved bi‐mesa structure has good separation from the mounting influences. © 2012 Wiley Periodicals, Inc. Electron Comm Jpn, 95(9): 54–60, 2012; Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/ecj.11385
Model‐based defect detection in multidimensional vector spaces
Authors: Honda Toshifumi , Obara Kenji , Harada Minoru , Igarashi Hajime 
Published: August 2012

Abstract222222222222222A highly sensitive inspection algorithm is proposed that extracts defects in multidimensional vector spaces from multiple images. The proposed algorithm projects subtraction vectors calculated from test and reference images to control the noise by reducing the dimensionality of vector spaces. The linear projection vectors are optimized using a physical defect model, and the noise distribution is calculated from the images. Because the noise distribution varies with the intensity or texture of the pixels, the target image is divided into small regions and the noise distribution of the subtraction images are calculated for each divided region. The bidirectional local perturbation pattern matching (BD‐LPPM), which is an enhanced version of the LPPM, is proposed to increase the sensitivity when calculating the subtraction vectors, especially when the reference image contains more high‐frequency components than the test image. The proposed algorithm is evaluated using defect samples for three different scanning electron microscopy images. The results reveal that the proposed algorithm increases the signal‐to‐noise ratio by a factor of 1.32 relative to that obtained using the Mahalanobis distance algorithm. © 2012 Wiley Periodicals, Inc. Electron Comm Jpn, 95(9): 44–53, 2012; Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/ecj.11390
Optimization of the motion trajectory for standing from a seated position by considering muscular load based on electromyography
Authors: Kushida Daisuke , Asakura Yuki , Kitamura Akira 
Published: August 2012

AbstractStanding from a chair is an important task of daily living for physically handicapped people. In a rehabilitation center, a healthcare professional plans motion on the basis of experience and knowledge so that a patient may stand up with little load. Therefore, there is a problem that the plan is occasionally different with each healthcare professional. In this paper, a method for generation of a motion trajectory to stand from a seated position with little load by using a genetic algorithm (GA) is proposed. The human body is expressed as a three‐rigid‐link model. In the model, the ankle, the knee, and the waist are set as the joints. Electromyographic (EMG) measurements of the muscle driving each joint were made and a model relating each joint torque to the EMG was constructed using the ARX model. The motion trajectory to stand from a seated position was generated by using a GA with its evaluation function based on the constructed ARX model. The generated motion trajectory was evaluated by experimental work with eight healthy subjects. The effect of the proposed method was objectively verified by the subjects' EMGs. In addition, the subjective effect of the proposed method was verified by analysis of the variance of the subjects' impressions. © 2012 Wiley Periodicals, Inc. Electron Comm Jpn, 95(9): 36–43, 2012; Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/ecj.11395
A Policy Description and Its Execution Scheduling for Automated IT Systems Management
Authors: Kudo Yutaka , Morimura Tomohiro , Masuoka Yoshimasa , Komoda Norihisa 
Published: August 2012

AbstractIn order to execute multiple policies in a policy‐based automation system safely and efficiently, we propose and develop a concurrency control mechanism for policy‐based automation systems. This mechanism analyzes the logical structure of the target application system to identify the IT resources affected by the policy action, and locks the resources to prevent conflicts between multiple policies. An efficient preemptive scheduling scheme for multiple policy executions is also developed. © 2012 Wiley Periodicals, Inc. Electron Comm Jpn, 95(9): 27–35, 2012; Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/ecj.11399
Three‐way translocation (X;20;16)(p11;q13;q23) in essential thrombocythemia implicates NFATC2 in dysregulation of CSF2 expression and megakaryocyte proliferation
Authors: Vieira Luís , Vaz Andreia , Matos Paulo , Ambrósio Ana Paula , Nogueira Manuel , Marques Bárbara , Pereira Ana Marques , Jordan Peter , da Silva Maria Gomes 
Published: August 2012

AbstractEssential thrombocythemia (ET) is a myeloproliferative neoplasm essentially characterized by excessive production of platelets. Molecular pathogenesis of ET is linked in approximately half of the patients to intracellular cytokine signaling dysregulation as a result of thrombopoietin receptor or Janus kinase 2 (JAK2) mutations. However, genetic defects underlying cytokine transcription have not been associated with ET. Using molecular cytogenetics and whole‐genome array analyses, we uncovered a submicroscopic deletion at 20q13.2 in a JAK2V617F‐positive ET patient with an acquired complex chromosome translocation. The deletion encompassed the nuclear factor of activated T‐cells, cytoplasmic, calcineurin‐dependent 2 (NFATC2) gene that encodes a transcription factor involved in the regulation of hematopoietic cytokines. RNA interference‐mediated suppression of NFATC2 mRNA or pharmacological inhibition of NFATC2 protein with 11R‐VIVIT in cultured JAK2V617F‐positive SET‐2 megakaryocytes increased colony stimulating factor 2 (granulocyte‐macrophage) (CSF2) mRNA and promoted cell proliferation. Moreover, impairment of NFATC2–calcineurin interaction with 11R‐VIVIT further reduced the transcription of the NFATC2 gene. Antibody‐mediated neutralization of CSF2 cytokine in inhibitor‐treated cells prevented 11R‐VIVIT‐induced cell proliferation, indicating that impairment of NFATC2–calcineurin interaction promotes megakaryocyte proliferation through up‐regulation of CSF2 transcription. Our results suggest a model in which haplo‐insufficiency of NFATC2 cooperates with activation of the JAK–STAT signaling pathway in the pathogenesis of JAK2V617F‐positive ET with del(20q). These results further indicate that pathogenesis of ET may be linked to genetic defects of other transcription factor genes involved in the regulation of cytokine expression. © 2012 Wiley Periodicals, Inc.
Elevated histone acetylations in Müller cell contribute to inflammation: A novel inhibitory effect of minocycline
Authors: Wang Lei‐Lei , Chen Hong , Huang Kun , Zheng Ling 
Published: August 2012

AbstractInflammation plays important roles in the development of diabetic retinopathy (DR). How Müller cells contribute to DR‐related inflammation remains unclear. We hypothesized that under diabetic conditions, elevated histone acetylations in Müller cells contribute to the inflammatory response. In this study, significantly increased histone acetylations, elevated histone acetyltranferases levels, and decreased histone deacetylases levels were found in the retinas of diabetic rats. Elevated AcH3K9 and AcH3K18 were partially co‐stained with Müller cells on retinal sections by immunofluorescence staining. Consistently, high‐glucose (HG) treated rMC‐1 cells, a Müller cell line, also showed upregulation of acetylated histones, accompanied with the overexpression of GFAP, p‐STAT3, and NFκB‐p65, and two inflammatory genes, TNFα and MCP‐1. Meanwhile, sodium butyrate (NaB)‐induced upregulation of acetylated histones is also accompanied with transcription of inflammatory genes. Minocycline, a drug with beneficial effects on DR, was found to downregulate HG‐induced Müller cell activation, inflammation, and acetylated H3K18 bound to the promoters of GFAP and inflammatory genes by chromatin immunoprecipitation assay. Furthermore, the effects of minocycline on HG‐induced elevation in histone acetylations were also demonstrated in isolated primary rat Müller cells. These findings suggest the elevation of histone acetylations in Müller cells plays important regulating roles in the inflammatory response during diabetic conditions. Inhibition of histone acetylation by minocycline is a novel function that may contribute to its beneficial effects on DR. © 2012 Wiley Periodicals, Inc.