1,724,721 research outputs found
Luciferase (<i>luc</i>) activity assay of <i>miR-3906</i> co-injected with plasmid constructs containing 3′UTR segment of putative <i>miR-3906</i>-target genes.
No full text<p>(<b>A</b>) Constructs for examining <i>luc</i> assay. The complete 3′UTR segments of <i>col1a2</i>, <i>dnajc10</i>, <i>homer-1b</i>, <i>six1a</i> and <i>trmt2a,</i> which are putative target genes of <i>miR-3906,</i> were ligated into the downstream of <i>luc</i> reporter gene contained in plasmid phRG-TK. (<b>B</b>) Plasmid pCMV-RFP-<i>miR-3906</i> [<i>miR-3906</i> (+)], a RFP reporter fused with pre-<i>miR-3906</i> and driven by CMV promoter, was co-transfected with pGL3-TK (internal control) and each examined construct, as indicated, into HEK 293T cells. Injection of pGL3-TK and each examined construct without containing pCMV-RFP-<i>miR-3906</i> [<i>miR-3906</i> (−)] served as a control group, and its <i>luc</i> activity was 100%. In zebrafish embryos, we co-injected synthetic pre-<i>miR-3906</i> RNA [miR-3906 (+)], pGL3-TK (internal control) and each examined construct in the experimental group. Injection of pGL3-TK and each examined construct without containing pre-<i>miR-3906</i> RNA [miR-3906 (−)] served as control group, and its <i>luc</i> activity was 100%. Data were presented as means±SD from three independent experiments (n = 3). (<b>C</b>) Injection of plasmid phRG-TK, in which <i>luc</i> expression was driven by thymidine kinase (TK), served as a control, and its <i>luc</i> activity was 100%. The relative <i>luc</i> expression level of each combination, as indicated, was examined. All data were presented as means±SD from three independent experiments (n = 3). (<b>D</b>) Various lengths of 3′UTR segment derived from <i>homer-1b</i> mRNA (from 1198 to 2222 nt) fused with <i>luc</i> reporter gene and driven by TK promoter were constructed as indicated. Plasmid alone or plasmid plus pre-<i>miR-3906</i> were individually injected in zebrafish embryos to perform <i>luc</i> assay. Data were presented as means±SD from three independent experiments (n = 3). Student's <i>t</i>-test determined significant differences between each group, and * indicates that the difference was significant at <i>P</i><0.05. (black box: <i>miR-3906-</i>target sequences; cross filled box: <i>miR-3906</i>-target mutated sequences).</p
Defective phenotypes induced by injection of excessive <i>miR-3906</i> and <i>homer-1b</i>-MO were similar.
No full text<p>(<b>A</b>–<b>I</b>) Embryos derived from transgenic line Tg (α-actin:RFP), in which red fluorescence protein (RFP) is expressed specifically in skeletal muscles, were injected with <i>miR-3906</i>-MO, <i>miR-3906</i> dsR (mature double- strand <i>miR-3906</i>), <i>homer-1b</i>-MO or <i>homer-1b</i> mRNA, as indicated. After injection, the development of trunk muscle was observed at 32 hpf. Compared with WT (<b>A</b>), the morphological trait of embryos injected with either <i>miR-3906</i>-MO (<b>B</b>) or <i>homer-1b</i> mRNA (<b>C</b>) was similar to that of WT. Embryos injected with <i>homer-1b</i>-MO (<b>D</b>–<b>F</b>) and exogenous <i>miR-3906</i> (<i>miR-3906</i> dsR) (<b>G</b>–<b>I</b>) displayed such defective phenotypes as body axis bending and trunk muscle shortening in tail. Based on the definition of three defective levels of muscle (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070187#s2" target="_blank">Materials and Methods</a>), we calculated the defective percentages of embryos injected with <i>homer-1b</i>-MO alone (<b>J</b>) and <i>homer-1b</i>-MO plus wobble <i>homer-1b</i> mRNA (<b>K</b>). The occurrence percentages of severe and mild defects of embryos injected with 1.15 ng <i>miR-3906</i> dsR alone, 1.15 ng <i>miR-3906</i> dsR plus 3 ng <i>homer-1b</i>-MO and 1.15 ng <i>miR-3906</i> dsR plus 400 pg <i>homer-1b</i> mRNA were calculated (<b>L</b>). n: indicates the total number of embryos after three injections. (<b>M</b>) Western blot analysis of Homer-1b (48 kD, arrow) in WT (lanes 1 and 4), 3 ng <i>homer-1b</i>-MO-injected embryos (lane 2), 8 ng <i>miR-3906</i>-MO-injected embryos (lane 3) and 1.15 ng <i>miR-3906</i> dsR-injected embryos (lane 5). Detection of α-tubulin served as internal control (arrowhead). The relative intensity of Homer-1b protein among WT, <i>homer-1b</i>-MO, <i>miR-3906</i>-MO and <i>miR-3906</i> dsR was also indicated.</p
[Veto of H. 3906, R-195]
No full textThis veto message from Governor Mark Sanford vetoes H. 3906, R-195, a bill seeks to expand the pool of students eligible for the LIFE or Palmetto Fellows Scholarships, or to relax the standards to be met by students in order to retain these scholarships
MicroRNA-3906 regulates fast muscle differentiation through modulating the target gene homer-1b in zebrafish embryos.
No full textA microRNA, termed miR-In300 or miR-3906, suppresses the transcription of myf5 through silencing dickkopf-related protein 3 (dkk3r/dkk3a) during early development when myf5 is highly transcribed, but not at late stages when myf5 transcription is reduced. Moreover, after 24 hpf, when muscle cells are starting to differentiate, Dkk3a could not be detected in muscle tissue at 20 hpf. To explain these reversals, we collected embryos at 32 hpf, performed assays, and identified homer-1b, which regulates calcium release from sarcoplasmic reticulum, as the target gene of miR-3906. We further found that either miR-3906 knockdown or homer-1b overexpression increased expressions of fmhc4 and atp2a1 of calcium-dependent fast muscle fibrils, but not slow muscle fibrils, and caused a severe disruption of sarcomeric actin and Z-disc structure. Additionally, compared to control embryos, the intracellular calcium concentration ([Ca(2+)]i) of these treated embryos was increased as high as 83.9-97.3% in fast muscle. In contrast, either miR-3906 overexpression or homer-1b knockdown caused decreases of [Ca(2+)]i and, correspondingly, defective phenotypes in fast muscle. These defects could be rescued by inducing homer-1b expression at later stage. These results indicate that miR-3906 controls [Ca(2+)]i homeostasis in fast muscle through fine tuning homer-1b expression during differentiation to maintain normal muscle development
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe 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
Moving to opportunity, leaving behind what? Evaluating the initial effects of a migration policy on incomes and poverty in source areas
No full textMigration to New Zealand and consequent remittance inflows are dominant features of many Pacific Island countries. Evaluating the effect of these people and money flows on incomes and poverty in the Pacific is potentially complicated by the non-random selection of emigrants. This paper uses the randomization provided by an immigration ballot under the Pacific Access Category (PAC) of New Zealand’s immigration policy to address this problem. We survey applicants to the 2002-05 PAC ballots in Tonga and compare outcomes for the remaining family of emigrants with those for similar families who were unsuccessful in the ballots. We then contrast these estimates with more conventional ones that construct no-emigration counterfactuals by deducting remittance income from the remaining family of PAC emigrants and adding back the potential home earnings of emigrants. The results suggest that the economic welfare of remaining family may fall in the initial period after members of their household move to New Zealand. We also find that non-experimental methods of constructing counterfactual income are likely to work well only in rare situations where there is random selection of emigrants
Variations on the Author
Full text link“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Change of <i>miR-3906</i> level affected the amount of <i>homer-1b</i> mRNA in zebrafish embryos.
No full text<p>(<b>A</b>, <b>D</b>) Compared with WT, injection of <i>miR-3906</i>-MO increased the expression level of <i>homer-1b</i> mRNA in embryos at 24 hpf (<b>A </b><i>vs</i>. <b>B</b>) and 32 hpf (<b>D </b><i>vs</i>. <b>E</b>). Injection of <i>miR-3906</i> dsR (mature double-strand <i>miR-3906</i>) reduced the expression level of <i>homer-1b</i> mRNA in the front of trunk muscle and was absent in the tail region at 24 hpf (<b>A </b><i>vs</i>. <b>C</b>) and 32-hpf (<b>D </b><i>vs</i>. <b>F</b>). (<b>G</b>) The expression levels of <i>homer-1b</i> in WT embryos, <i>miR-3906</i>-MO-injected embryos and <i>miR-3906</i> dsR-injected embryos at 24 hpf and 32 hpf were quantified. Each one was carried out with 100 embryos, and triplicate experiments were performed (n = 3). The numbers shown in the lower-right corner of panels A–F indicate the number of phenotypes out of the total number of embryos examined. SigmaPlot software was used to perform Student’s <i>t</i>-test. *: indicates the difference at <i>P<0.05</i> level; **: indicates the difference at <i>P<0.01</i> level.</p
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