118,179 research outputs found
Association of RyDEN with DENV RNA.
No full text<p>(A) HepG2 cells expressing V5-RyDEN (WT and NLS-L mutant) or V5-DHFR (control) were infected with DENV-2 at an MOI of 5, and 6 h after infection, cell lysates were subjected to RIP assay using anti-V5 antibodies. A portion of the cell lysates were used for total RNA extraction, and DENV RNA was quantified by qRT-PCR, which was normalized with GAPDH mRNA. (B) Immunoblotting analysis to detect V5-tagged proteins. Portions of the immunoprecipitated (IP, top panel) and input (middle and bottom panels) fractions were subjected to immunoblotting (IB) analysis using anti-V5 (for RyDEN and RLuc, top and middle panels) and anti-actin (bottom panel) antibodies. (C) Level of DENV RNA in immunoprecipitates. Total RNA was extracted from immunoprecipitated samples, and the DENV RNA level was analyzed by qRT-PCR. (D) Schematic diagram of AlphaScreen assay to detect the binding of RyDEN and DENV 3’UTR RNA. FLAG-tagged RyDEN produced by the wheat germ cell-free system was incubated with biotin-labelled (biotinylated) DENV-2 3’UTR RNA in the presence of GST-tagged proteins. RyDEN and 3’UTR RNA interaction bridges the streptavidin (sa)-coated donor bead and anti-FLAG-conjugated acceptor bead via recognition of biotin (b) of RNA and N-terminal FLAG-tag (f) of protein, respectively. Upon excitation at 680 nm, single oxygen molecules (<sup>1</sup>O<sub>2</sub>) are produced from the donor beads, which react with the acceptor beads, resulting in light emission measured between 520 and 620 nm (AlphaScreen signals). (E) <i>In vitro</i> interaction of RyDEN and DENV RNA. The AlphaScreen-based RNA-binding assay was performed with 20 nM FLAG-tagged proteins (RyDEN WT [Rxns 1, 2, 3, 5, and 6], RyDEN NLS-L mutant [Rxn 7], or DHFR [Rxn 4]) and 3.5 ng/μl substrate RNA (unlabeled DENV 3'UTR [Rxn 1], biotinylated control [derived from DHFR gene, 480 base, Rxns 2 and 3], or biotinylated 3'UTR [Rxns 4–7] RNA) in the presence of 20 nM GST-tagged proteins (RABPC1 [Rxns 3, 6, and 7] or DHFR [Rxns 1, 2, 4, and 5]). Statistical significance was determined by one-way ANOVA with Dunnett’s multiple comparison test. ns, no significance.</p
Requirement of PABPC1 interaction domain for anti-DENV activity of RyDEN.
No full text<p>(A) Schematic of full-length (WT) RyDEN and its mutants used in experiments. N-terminal (51–291, 101–291, and 151–291) and C-terminal (1–250) deletion mutants and a site-directed mutant, in which arginine (R) and lysine (K) residues in a putative NLS sequence (NLS-L) were substituted with alanine (A), were constructed as V5-tagged proteins. Predicted α-helix (black) and β-sheet (gray) regions are also shown. (B) Mapping of the PABPC1-binding domain in RyDEN. A series of V5- RyDEN, including its truncation mutants (lanes 2–6) and V5-RLuc (control, lane 1), were lentivirally expressed in Huh7.5 cells, and cell lysates (input, bottom panel) were subjected to co-immunoprecipitation (IP) analysis using anti-V5 antibodies. Immunoprecipitates were then analyzed by immunoblotting (IB) using anti-V5 (for RyDEN and RLuc, top panel) or anti-PABPC1 (middle panel). Masses of molecular weight standards are indicated at left. (C) IFA of V5-protein expressing cells. HepG2 cells expressing V5-RyDEN or parental HepG2 cells were fixed with 4% paraformaldehyde (PFA), permeabilized with 0.1% Triton X-100, blocked with 5% goat serum, and stained with an anti-V5 antibody, followed by detection with Alexa Fluor 488-conjugated anti-mouse secondary antibody (top row). (D) IFA for endogenously expressed RyDEN. HepG2 cells that had been cultured in the presence or absence of IFN-α/ω (1,000 units/ml) for 24 h were fixed with PFA, permeabilized with 1% Triton X-100, blocked with Blocker Casein (Thermo Scientific), and stained with anti-RyDEN rabbit serum, followed by detection with FITC-conjugated anti-rabbit secondary antibody (top row). As another controls, V5-RyDEN- or V5-DHFR-expressing HepG2 cells was also stained with the anti-RyDEN serum (third and fourth columns). (E) Localization of RyDEN deletion mutants. Huh7.5 cells expressing V5-RyDEN (WT), its truncation mutants (1–250, 51–291, 101–291, 151–291), or control V5-RLuc were subjected to IFA using anti-V5 antibody. In all IFA, cell nuclei were stained with DAPI (center rows), and merged images are shown in the bottom rows. (F) HepG2 cells stably expressing V5- RyDEN (WT and NLS-L mutant) or V5-RLuc were generated by lentiviral vector transduction, and cell lysates (input) were used for immunoprecipitation analysis using anti-V5 antibodies. V5-tagged proteins and PABPC1 in the immunoprecipitates were detected by immunoblotting using anti-V5 (for RyDEN and RLuc, top panel) and anti-PABPC1 (middle panel) antibodies. (G) Activity of the RyDEN NLS-L mutant in the suppression of DENV replication. V5-tagged protein-expressing HepG2 cells were infected with DENV-2 at an MOI of 1, and culture supernatants were subjected to plaque assay 2 days after infection to measure the virus titer. Statistical significance was determined by one-way ANOVA with Dunnett’s multiple comparison test.</p
Rationale, design, and baseline characteristics for a large international trial of cardiovascular disease prevention in people with dysglycemia: the ORIGIN Trial (Outcome Reduction with an Initial Glargine Intervention)
No full textAIMS: Impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and diabetes arise due to insufficient insulin secretion and are risk factors for cardiovascular (CV) events. Thus, targeting normal fasting glucose levels with insulin may reduce CV events. Previous studies suggest that omega-3 fatty acid supplements may reduce CV death; however, their effect in high-risk dysglycemic individuals is not known.
METHODS: People aged > or = 50 years with evidence of CV disease and with IFG, IGT, newly detected or established diabetes (on 0 or 1 oral agent), and a local glycated hemoglobin < 150% of the upper limit of normal for that assay were recruited and allocated to (a) either 1 daily injection of insulin glargine with the dose titrated to achieve a fasting plasma glucose < or = 5.3 mmol/L (95 mg/dL), or standard glycemic care; and (b) either omega-3-acid ethyl esters 90 (1 g consisting of EPA 465 mg and DHA 375 mg) or identical placebo, according to a 2 x 2 factorial design. The 2 different primary outcomes for the insulin and omega-3 fatty acid arms are CV events and CV death, respectively.
RESULTS: A total of 12,612 (mean age 64, 35% women) people in 40 countries were randomized during a 2-year period ending December 2005. Eighty-two percent had established diabetes, 6% had new diabetes, and 12% had IGT or IFG; the mean fasting plasma glucose was 7.3 mmol/L (131 mg/dL).
CONCLUSIONS: The ORIGIN trial will determine whether or not either or both of these interventions can reduce CV events
Going Beyond Counting First Authors in Author Co-citation Analysis
No full textThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Square Dancing with the Stars to Enhance Dynamic Hirschman Linkages?
No full textIn this Presidential Address, the author takes the reader on a reconnaissance of his life and time as a regional scientist. He points out scenery he found scintillating along the way, hoping that some may pick up the banner and chew on a few of the ideas for a while. He suggests a revisit to Albert O. Hirschman’s notion of key sectors and more empirical analysis related to Marcus Berliant’s and Masahisa Fujita’s notion of knowledge creation and transfer.Presidential Address, San Antonio, Texas, March 29, 2014 (53rd Meetings of the Southern Regional Science Association
Appropriate Similarity Measures for Author Cocitation Analysis
No full textWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
Letter from unknown writer to Jesse L. Boyce
No full textLetter to Jesse L. Boyce from unknown author (possibly Jack) about the investigation into the powder magazine located in the Grand Canyon. Some personal news is included in the letter such as the writer's marriage to the daughter of C.A. Taylor, former Supervisor of Cochise County
- …
