168 research outputs found

    Aerodynamic forces on two-dimensional rectangular cylinders subjected to accelerating flow: effect of side ratio

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    This paper was reviewed and accepted by the APCWE-IX Programme Committee for Presentation at the 9th Asia-Pacific Conference on Wind Engineering, University of Auckland, Auckland, New Zealand, held from 3-7 December 2017

    Alignment of potential NES sequences of L-periaxin with previously characterized leucine-rich NESs.

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    <p>NES sequences in mitogen-activated protein kinase kinase (MAPKK), zyxin, HIV.Rev, c-Abl, and LIM-kinase 1 are obtained from previous studies <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0091953#pone.0091953-Yang1" target="_blank">[22]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0091953#pone.0091953-Fischer1" target="_blank">[29]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0091953#pone.0091953-FukudaM1" target="_blank">[30]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0091953#pone.0091953-NixDA1" target="_blank">[31]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0091953#pone.0091953-Taagepera1" target="_blank">[32]</a>. Residue numbers are indicated in parentheses. Consensus sequence is shown at the bottom. Ψ indicates hydrophobic residues, which include leucine, isoleucine, valine, phenylalanine, and methionine.</p

    Nuclear accumulation of L-periaxin by mutating Leu<sub>83</sub> and Leu<sub>85</sub> to Gln.

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    <p>(A) Structures of L-PRX, PDZ-cyclin A1, and their mutants. Gray boxes indicate the amino acid residues replaced with Gln in the PDZ domain. (B) RSC96 cells were transfected with the plasmids encoding EGFP-tagged L-PRX, PDZ-cyclin A1, and their mutants. Scale bar = 25 μm. (C) The cytoplasm and nucleus were separated and determined the distribution of protein in the two fractions by Western blotting. β-actin and Histone served as an internal control for cytosol and nucleus, respectively. (D) Quantitative analysis of the distribution of EGFP fusion proteins in the two fractions by normalized to the internal control level. *P<0.05.</p

    Quantification of microRNA by DNA–Peptide Probe and Liquid Chromatography–Tandem Mass Spectrometry-Based Quasi-Targeted Proteomics

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    The distorted and unique expression of microRNAs (miRNAs) in cancer makes them an attractive source of biomarkers. However, one of prerequisites for the application of miRNAs in clinical practice is to accurately profile their expression. Currently available assays normally require pre-enrichment, amplification, and labeling steps, and most of them are semiquantitative. In this study, we converted the signal of target miR-21 into reporter peptide by a DNA-peptide probe and the reporter peptide was ultimately quantified using LC-MS/MS-based targeted proteomics. Specifically, substrate peptide GDKAVLGVDPFR containing reporter peptide AVLGVDPFR and tryptic cleavage site (lysine at position 3) was first designed, followed by the conjugation with DNA sequence that was complementary to miR-21. The newly formed DNA-peptide probe was then hybridized with miR-21, which was biotinylated and attached to streptavidin agarose in advance. After trypsin digestion, the reporter peptide was released and monitored by a targeted proteomics assay. The obtained limit of quantification (LOQ) was 1 pM, and the detection dynamic range spanned ∼5 orders of magnitude. Using this assay, the developed quasi-targeted proteomics approach was applied to determine miR-21 level in breast cells and tissue samples. Finally, qRT-PCR was also performed for a comparison. This report grafted the strategy of targeted proteomics into miRNA quantification

    Cytoplasmic localization of a normal nuclear cyclin A1 by fusing with the PDZ domain of L-periaxin.

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    <p>(A) Schematic representation of cyclin A1 and its chimeric proteins fused with the PDZ domain of L-periaxin. N-terminal hatched boxes indicate EGFP-tagged peptides. The plain numbers on top of the boxes indicate the amino acid residue number of cyclin A1. Italic numbers correspond to the residues flanking the PDZ-domain of L-periaxin. The subcellular localization of each protein, when expressed in RSC96 cells, is indicated on the right. N, nucleus, C, cytoplasm. (B) Fluorescence analysis of RSC96 cells transfected with the plasmids encoding EGFP, EGFP-cyclin A1, and EGFP-PDZ-cyclin A1. Scale bar = 25 μm. (C) GFP fusion proteins are stable in the cell which is shown by Western blotting.</p

    PDZ is necessary to accomplish an efficient nuclear export of L-PRX.

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    <p>(A) Structures of L-PRX and its PDZ-deleted mutants (DPDZ-L-PRX). Amino acid residues flanking each domain are numbered on top of the diagram. (B) Subcellular localization of EGFP-tagged L-PRX and its PDZ-deleted mutant (DPDZ-L-PRX). RSC96 cells were transfected with the plasmids encoding EGFP-tagged full-length L-PRX and its DPDZ-L-PRX. Scale bar = 25 μm. (C) GFP fusion constructs detected by Western blotting to ensure that the visualized proteins are not free GFP, or GFP fused to a truncated protein.</p

    Crystal Growth of Hydrogen/Tetra‑<i>n</i>‑butylammonium Bromide Semiclathrates Based on Morphology Study

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    Morphology studies were conducted for the first time on the mixed hydrogen/tetrabutylammonium bromide (TBAB) semiclathrates. Morphology study dealt with the observation of nucleation of the first hydrate crystal, the growth of hydrates and the characteristic appearance of crystals through the microscope. Morphology changes occurring during the formation of mixed hydrogen hydrates using TBAB as a promoter were observed through the microscope. Concentration of TBAB was varied and the influence of concentration on the hydrate crystal morphology was studied. At lower TBAB concentration (1 and 2 mol %) needle like and equiaxial crystals were formed initially which later developed into columnar crystals having high transmittance and retained their individual structure. However, at higher TBAB concentration (2.5 mol % and above), cylinder like crystals were observed which developed into dense irregular shaped crystals with lower transmittance. The individual crystals cannot be distinguished, they become mushy and grow as layers. Characteristic wave formations were observed during crystal formation in all the cases. With the decrease in subcooling or with the increase in experimental pressure (driving force), the dimensions of the formed crystals was initially same but later increased to around 2–4 times the original size

    Simultaneous Quantification of Protein Phosphorylation Sites using Liquid Chromatography–Tandem Mass Spectrometry-Based Targeted Proteomics: A Linear Algebra Approach for Isobaric Phosphopeptides

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    As one of the most studied post-translational modifications (PTM), protein phosphorylation plays an essential role in almost all cellular processes. Current methods are able to predict and determine thousands of phosphorylation sites, whereas stoichiometric quantification of these sites is still challenging. Liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS)-based targeted proteomics is emerging as a promising technique for site-specific quantification of protein phosphorylation using proteolytic peptides as surrogates of proteins. However, several issues may limit its application, one of which relates to the phosphopeptides with different phosphorylation sites and the same mass (i.e., isobaric phosphopeptides). While employment of site-specific product ions allows for these isobaric phosphopeptides to be distinguished and quantified, site-specific product ions are often absent or weak in tandem mass spectra. In this study, linear algebra algorithms were employed as an add-on to targeted proteomics to retrieve information on individual phosphopeptides from their common spectra. To achieve this simultaneous quantification, a LC–MS/MS-based targeted proteomics assay was first developed and validated for each phosphopeptide. Given the slope and intercept of calibration curves of phosphopeptides in each transition, linear algebraic equations were developed. Using a series of mock mixtures prepared with varying concentrations of each phosphopeptide, the reliability of the approach to quantify isobaric phosphopeptides containing multiple phosphorylation sites (≥2) was discussed. Finally, we applied this approach to determine the phosphorylation stoichiometry of heat shock protein 27 (HSP27) at Ser78 and Ser82 in breast cancer cells and tissue samples

    Synthesis of Hyperbranched Perovskite Nanostructures

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    Complex functional perovskite oxides find use in a wide range of applications as dielectric, electronic, photocatalytic, optical, and ion-conducting materials. The ability to synthesize perovskites in hierarchical structures can allow for new properties or phenomena not observable in bulk morphologies. Herein we report on the solution-phase synthesis of cubic perovskite potassium lanthanum titanate into orthogonal nanostructures with hyperbranched and hexapod morphology through a facile hydrothermal synthesis without using a template, catalyst, substrate, or structure-directing agent. A systematic study of the reaction conditions and role of precursors was performed and a growth mechanism based on homogeneous dissolution–precipitation was determined

    The Development and the Role of International Criminal Justice in Today's World

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    This thesis deals with international criminal justice, which began to influence international politics during the last twenty years. After the end of the Cold War, a lot of armed conflicts were breaking out. They were accompanied by unprecedented inhuman acts and atrocities. The international community had to find a solution for how to respond to such events. In 1993, the United Nations Security Council acted under Chapter VII of The Charter of the United Nations and decided unanimously upon the establishing of an ad hoc International Criminal Tribunal for the Former Yugoslavia. The Tribunal's role was to prosecute persons responsible for serious violations of international humanitarian law committed during the Balkans conflict. A year later, in 1994, the Security Council decided to establish another ad hoc tribunal - The International Criminal Tribunal for Rwanda, which served to punish the architects of Rwandan genocide. Both tribunals sped up negotiations and the decision to establish the permanent International Criminal Court, whose objective is to help end impunity for the perpetrators of the most serious crimes of concern to the international community. The crime of genocide, war crimes, crimes against humanity and in the future the crime of aggression. Firstly, this thesis analyses the ad..
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