Distinctive Profile of IsomiR Expression and Novel MicroRNAs in Rat Heart Left Ventricle

MicroRNAs (miRNAs) are single-stranded non-coding RNAs that negatively regulate target gene expression through mRNA cleavage or translational repression. There is mounting evidence that they play critical roles in heart disease. The expression of known miRNAs in the heart has been studied at length by microarray and quantitative PCR but it is becoming evident that microRNA isoforms (isomiRs) are potentially physiologically important. It is well known that left ventricular (patho)physiology is influenced by transmural heterogeneity of cardiomyocyte phenotype, and this likely reflects underlying heterogeneity of gene expression. Given the significant role of miRNAs in regulating gene expression, knowledge of how the miRNA profile varies across the ventricular wall will be crucial to better understand the mechanisms governing transmural physiological heterogeneity. To determinine miRNA/isomiR expression profiles in the rat heart we investigated tissue from different locations across the left ventricular wall using deep sequencing. We detected significant quantities of 145 known rat miRNAs and 68 potential novel orthologs of known miRNAs, in mature, mature* and isomiR formation. Many isomiRs were detected at a higher frequency than their canonical sequence in miRBase and have different predicted targets. The most common miR-133a isomiR was more effective at targeting a construct containing a sequence from the gelsolin gene than was canonical miR-133a, as determined by dual-fluorescence assay. We identified a novel rat miR-1 homolog from a second miR-1 gene; and a novel rat miRNA similar to miR-676. We also cloned and sequenced the rat miR-486 gene which is not in miRBase (v18). Signalling pathways predicted to be targeted by the most highly detected miRNAs include Ubiquitin-mediated Proteolysis, Mitogen-Activated Protein Kinase, Regulation of Actin Cytoskeleton, Wnt signalling, Calcium Signalling, Gap junctions and Arrhythmogenic Right Ventricular Cardiomyopathy. Most miRNAs are not expressed in a gradient across the ventricular wall, with exceptions including miR-10b, miR-21, miR-99b and miR-486.

Expression of the counter-regulatory peptide intermedin is augmented in the presence of oxidative stress in hypertrophied cardiomyocytes.

Background: Intermedin (IMD), a novel cardiac peptide related to adrenomedullin (AM), protects against myocardial ischemia-reperfusion injury and attenuates ventricular remodelling. IMD’s actions are mediated by a calcitonin receptor-like receptor in association with receptor activity modifying proteins (RAMPs 1-3). Aim/method: using the spontaneously hypertensive rat (SHR) and normotensive Wistar Kyoto (WKY) rat at 20 weeks of age, to examine (i) the presence of myocardial oxidative stress and concentric hypertrophy; (ii) expression of IMD, AM and receptor components. Results: In left and right ventricular cardiomyocytes from SHR vs. WKY cell width (26% left, 15% right) and mRNA expression of hypertrophic markers ANP (2.7 fold left, 2.7 fold right) and BNP (2.2 fold left, 2.0 fold right) were enhanced. In left ventricular cardiomyocytes only (i) oxidative stress was indicated by increased membrane protein carbonyl content (71%) and augmented production of O2- anion (64%); (ii) IMD (6.8 fold), RAMP1 (2.5 fold) and RAMP3 (2.0 fold) mRNA was increased while AM and RAMP2 mRNA was not altered; (iii) abundance of RAMP1 (by 48%), RAMP2 (by 41%) and RAMP3 (by 90%) monomers in cell membranes was decreased. Conclusion: robust augmentation of IMD expression in hypertrophied left ventricular cardiomyocytes indicates a prominent role for this counter-regulatory peptide in the adaptation of the SHR myocardium to the stresses imposed by chronic hypertension. The local concentration and action of IMD may be further enhanced by down-regulation of NEP within the left ventricle.

Myocardial infarction in the C57BL/6J mouse - A quantifiable and highly reproducible experimental model

Introduction: The laboratory mouse is a powerful tool in cardiovascular research. In this report, we describe a method for a reproducible mouse myocardial infarction model that would allow subsequent comparative and quantitative studies on molecular and pathophysiological variables. Methods: (A) The distribution of the major coronary arteries including the septal artery in the left ventricle of the C57BL/6J mice (n=20) was mapped by perfusion of latex dye or fluorescent beads through the aorta. (B) The territory of myocardial infarction after the ligation of the most proximal aspect of the left anterior descending (LAD) coronary artery was quantified. (C) The consistency in the histological changes parallel to the infarction at different time points was analyzed. Results: (A) The coronary artery tree of the mouse is different from human and, particularly, in regard to the blood supply of the septum. (B) Contrary to previous belief, the septal coronary artery in the mouse is variable in origin. (C) A constant ligation of the LAD immediately below the left auricular level ensures a statistically significant reproducible infarct size. (D) The ischemic changes can be monitored at a histological level in a way similar to what is described in the human. Conclusion: We illustrate a method for maximal reproducibility of experimental acute myocardial infarction in the mouse model, due to a consistent loss of perfusion in the lower half of the left ventricle. This will allow the study of molecular and physiological variables in a controlled and quantifiable experimental model environment. (C) 2004 Elsevier Inc. All rights reserved.

Key issues in the study of new and alternative social movements in Spain: The Left, Identity and Globalizing Processes

This article highlights the importance of dedicating a whole special issue on New and Alternative Social movements in Spain. It sets the basis for this endeavour by emphasizing the importance of the 2004, unexpected, electoral victory of the Spanish socialists, and the subsequent satisfaction of the important demands promoted by certain social movements actors and Spanish society in general (the withdrawal of Spanish troops from Iraq, the cancellation of the National Hydrological Plan and the Legalization of same sex marriages. The view supported is that these developments signify the end of a protest cycle, which could have the same effect with the early 1980s socialist victory. After a discussion around the low associationalism that characterizes Spanish society and recent experience of authoritarianism, it is suggested that it is time for the study of new and alternative social movements in Spain and other south European societies to move beyond the emphasis on exceptionality but appreciate differences by focusing on the available political opportunities and the identity of social movement actors. The remainder of the article is dedicated to introducing the contributing articles.

Design strategies for dual-band class-e power amplifier using composite right/left-handed transmission lines

In this article we propose a technique for dual-band Class-E power amplifier design using composite right/left-handed transmission lines, CRLH TLs. Design equations are presented and design procedures are elaborated. Because of the nonlinear phase dispersion characteristic of CRLH TLs, the single previous attempt at applying this method to dual bond Class-E amplifier design was not sufficient to simultaneously satisfy, the minimum requirement of Class-E impedances at both the fundamental and the second harmonic frequencies. This article rectifies this situation. A design example illustrating the synthesis procedure for a 0.5W-5V dual band Class-E amplifier circuit simultaneously operated at 900 MHz and 2.4 GHz is given and compared with ADS simulation.

Right-left discrimination among medical students: Getting it right all of the time?

Objective To determine medical students’ self awareness and ability to discriminate right from left; to identify characteristics associated with this ability; and to identify any techniques used to aid discrimination. Design Questionnaire and psychometric study. Setting Undergraduate medical school, Northern Ireland. Participants 290 first year undergraduate students. Main outcome measure Medical students’ ability to discriminate right from left using the Bergen right-left discrimination test. Results Test scores ranged from 31 to 143 on a scale of 0- 144 (mean 112 (standard deviation 22.2)). Male students significantly outperformed female students (117.18 (26.96) v 110.80 (28.94)). Students who wanted to be surgeons performed significantly better than those who wanted to be general practitioners or medical doctors (119.87 (25.15) v 110.55 (27.36) v 112.50 (26.88)). The interaction effect for sex and career wishes was not significant (P=0.370). Students who used learnt techniques to help them discriminate scored significantly less than those who did not (P

Exile or emigrant? Female ex-prisoners who left Ireland 1850-1921

In the popular mind, the concept of 'emigration' usually refers to people voluntarily leaving one country to go to another in search of a new and better life. It presupposes some degree of choice, although it is accepted that for many emigrants, such as those who left Ireland during the nineteenth century, there were few incentives to stay at home. Current scholarship on voluntary and forced movements of people demonstrates that the distinction between the categories of 'voluntary emigrant' and 'forced exile' is often blurred. Orm Overland's study of refugee communities in the United States highlights the fact that, although the differences between the 'emigrant' and the 'exile' may be clear in extreme cases, this is not always true, as there may be 'pressing political or economic reasons behind a decision to emigrate'. Migration scholars Jan Lucassen and Leo Lucassen also question the adequacy of conceptual models of migration based on what Lindsay Proudfoot and Dianne Hall refer to as the 'straightforward binarism between free and unfree emigration'. The questions raised by these scholars are very relevant to the study of Irish people who left their country during the second half of the nineteenth century immediately after they had been discharged from prison or from Dundrum. Their stories are discussed here against a background of substantial scholarship on emigration from Ireland and on the criminal justice system within Ireland. According to David Fitzpatrick, at least eight million men, women and children emigrated from Ireland between 1801 and 1921. This large-scale movement of people was generally characterised by the voluntary emigration of individuals who funded their own passages. However, it also included schemes of assisted emigration, funded variously by governments, landlords, the poor law authorities, earlier emigrants, and philanthropists. In addition, it included people who were transported from Ireland by means of the criminal justice system a practice that had originated in the seventeenth century. What is less well known is that after the end of transportation from Ireland to eastern Australia in 1853, to Bermuda in 1863 and to Western Australia in 1868, Irish convicts continued to be channelled towards emigration by being offered early release if they agreed to leave Ireland. These people, and especially the women among them, are the subject of this article.

Periodically loaded dipole array supporting left-handed propagation

Periodically loaded dipole arrays printed on grounded dielectric substrate are shown to exhibit left-handed propagation properties. In an equivalent transmission line representation, lefthandedness emerges due to the excess series capacitance and shunt inductance. Based on this concept, the authors study the distribution of the modal fields and the variation of series capacitance and shunt inductance as the dipoles are loaded with stubs. Full wave dispersion curves that show the gradual transition from a right-handed to a left-handed medium upon periodically loading the dipoles with stubs are presented. An equivalent circuit is derived that matches to a very good extent the full wave result. The cell dimensions are a small fraction of the wavelength (),15), so the structure can be considered as an equivalent homogeneous surface. The structure is simple, readily scalable to higher frequencies and compatible with low-cost fabrication techniques.