1,720,964 research outputs found
Regulation of Glut-4 Expression in Skeletal Muscle cells: The Roles of Nuclear Respiratory Factor-1 and calcium/calmodulin dependent protein Kinase
No full textGLUT4 protein is the major glucose transporter in skeletal muscle and is vital in the maintenance of euglycemia (17; 108). Underexpression of GLUT4 or impairement of its translocation from intracellular compartments to the cell surface, are linked to diminished glucose transport, hyperglycemia and type II diabetes (59; 61; 153). Type II diabetes can be alleviated by increasing GLUT4 expression (223). Previous reports have shown that overexpression of NRF-1 and activation of CaMKII increases GLUT4 expression but the mechanisms involved have not be characterized (10; 173). Therefore, the objective of this thesis was to investigate the molecular mechanisms by which NRF-1 and CaMK II regulate GLUT4 expression in C2C12 myocytes. We engineered C2C12 cells that overexpressed NRF-1 in response to doxycycline (Dox) using a Tet-On gene expression system and assessed the effects of NRF-1 overexpression on: a) MEF2A, GLUT4 and δALAS proteins by western blot, and b) the binding of NRF-1 to mef2a and δalas genes and MEF2A to the glut4 gene, by chromatin immunoprecipitation assay (ChIP). The importance of MEF2A in NRF-1-induced increase in GLUT4 expression was investigated by silencing MEF2A expression using small interference RNA (siRNA). CaMK II was activated in wild-type C2C12 myocytes using 10 mM caffeine and was inhibited by 25 μ M KN93. Acetylation of histones in the vicinity of NRF-1 and MEF2A binding sites on the mef2a and glut4 genes, respectively, were assessed by ChIP assay. HDAC5 nuclear export was assessed by immunocytochemistry and mRNA levels by qRT-PCR. Overexpression of NRF-1 resulted in ~3-fold increases in mef2a-bound NRF-1 and glut4 -bound MEF2A at 6 h and 8 h post Dox treatment, respectively. MEF2A and GLUT4 proteins were both increased ~1.6-fold at 6 h and 18 h post Dox treatment. Silencing of MEF2A caused a marked downregulation of GLUT4 expression in NRF-1-overexpressing cells
Involvement of p300 in caffeine-induced hyper-acetylation of histones at the MEF2 binding domain on the Glut4 gene
No full textIncludes abstract.
Includes bibliographical references
Over-expression of NRF-1 in C2C12 myotubes increases GLUT4 content via a transcriptional cascade involving MEF2A
No full textIncludes abstract.Includes bibliographical references.Previous studies have shown that over-expression of nuclear respiratory factor (NRF)-1 in mice increases glucose transporter (GLUT)-4 and myocyte enhancer factor (MEF-) 2A content, but the mechanisms have not been elucidated. Because NRF-1 has a binding site on the mef2a gene, and MEF2A binds the glut4 gene as a MEF2A-MEF2D heterodimer, the aims of this study were to determine whether NRF-1 over-expression a) enhanced GLUT4 expression indirectly via MEF2A and b) alters MEF2A-MEF2D dimer formation in C2C12 myotubes
Regulation of skeletal muscle glucose transporter 4 expression in fructose-fed exercised rats
No full textIncludes abstract.Includes bibliographical references.Several studies have found that the expression of skeletal muscle glucose transporter 4 (GLUT4) is decreased by high fructose consumption but increased by exercise. However, the amounts of fructose used in these studies were extremely high and the effects of moderate feeding protocols are not known. While it is known that exercise enhances GLUT4 expression via increased histone H3 acetylation and binding of the myocyte enhancer factor- 2A (MEF2A) transcription factor to its binding domain on the Glut4 gene promoter, the impact of fructose consumption on this interaction has not been studied. Moreover, there is no direct evidence that an increase in MEF2 binding is due to increased accessibility of the MEF2 binding domain to transcription factors. This study tested the hypothesis that both exercise and high fructose consumption affect GLUT4 expression by altering the accessibility of the MEF2 binding domain on the Glut4 gene promoter via remodelling of chromatin in that region. Male Wistar rats (n=30) were randomly assigned to three dietary groups: a) standard Chow, b) Chow + 10% fructose drink and c) Chow + 10% maltodextrin drink. All rats had access to drinking water and chow ad libitum for a period of 13 days. In the last 6 days of the experiment 5 animals in each group performed 3 x 17 min daily bouts of intermittent swimming, with a load equivalent to 5% bodyweight attached to their tails. The remaining 5 rats from each group were untrained. Animals were fasted overnight on the last day of the experiment, anaesthetized and sacrificed on the morning of day 14. Triceps muscle were harvested and used: (a) for measurement of total GLUT4 content by western blot, (b) to obtain nuclei for assessment of accessibility of a 350bp region encompassing the MEF2 element on the Glut4 gene using nuclease digestion assay, and (c) to measure the acetylation of histones H3 and bound MEF2A in the region above using chromatin immunoprecipitation (ChIP) assay. Blood was also collected and assayed for fasting serum glucose, insulin and free fatty acids
Exercise-induced protection against insulin resistance and type 11 diabetes : the role of calcium
No full textIncludes abstract.Includes bibliographical references (p. 89-106).Regular exercise protects individuals against developing insulin resistance and type II diabetes. This effect of exercise does not appear to be due to an improvement in the insulin signalling pathway but instead due to an increase in the content of the insulin-regulatable glucose transporter (GLUT4) in skeletal muscle (84). Understanding the mechanisms by which exercise increases GLUT4 levels in skeletal muscle may reveal targets for pharmaceuticals to treat insulin resistance and type II diabetes. Although in vitro binding assays have shown that GLUT4 expression during exercise is mediated by the binding of myocyte enhancer factor-2A (MEF2A) to its cis-element on the Glut4 promoter (122), this has not been demonstrated in vivo. Moreover, the mechanisms by which exercise increases MEF2A binding to the Glut4 promoter have not been fully characterised
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
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
Appropriate Similarity Measures for Author Cocitation Analysis
Full text linkWe 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
Dispelling the Myths Behind First-author Citation Counts
Full text linkWe conducted a full-scale evaluative citation analysis study of scholars in the XML research field to explore just how different from each other author rankings resulting from different citation counting methods actually are, and to demonstrate the capability of emerging data and tools on the Web in supporting more realistic citation counting methods. Our results contest some common arguments for the continued
use of first-author citation counts in the evaluation of scholars, such as high correlations between author rankings by first-author citation counts and other citation
counting methods, and high costs of using more realistic citation counting methods that are not well-supported by the ISI databases. It is argued that increasingly available digital full text research papers make it possible for citation analysis studies to go beyond what the ISI databases have directly supported and to employ more
sophisticated methods
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