Classical, novel and atypical isoforms of PKC stimulate ANF- and TRE/AP-1-regulated-promoter activity in ventricular cardiomyocytes

Cultured neonatal rat ventricular myocytes were co-transfected with expression plasmids encoding protein kinase C (PKC) isoforms from each of the PKC subfamilies (classical PKC-α, novel PKC-ε or atypical PKC-ξ) together with an atrial natriuretic factor (ANF) reporter plasmid. Each PKC had been rendered constitutively active by a single Ala→Glu mutation or a small deletion in the inhibitory pseudosubstrate site. cPKC-α, nPKC-ε or aPKC-ξ expression plasmids each stimulated ANF-promoter activity and expression of a reporter gene under the control of a 12-O-tetradecanoylphorbol 13-acetate-response element (TRE). Upregulation of the ANF promoter is characteristic of the hypertrophic response in the heart ventricle and a TRE is present in the ANF promoter. Thus all subfamilies of PKC may have the potential to contribute to hypertrophic response in cardiomyocytes

BIM’s role in improving the management and delivery of occupational safety and health and saving lives in construction

Published in BIM Coordinators Summit Magazine.Despite improvements and modernizations in construction processes, accidents are still persistent and have considerable financial and logistical impacts on projects. The UK HSE recently revealed that there were 145 work-related deaths in 2022/23 with the highest number of 45 occurring in construction. Research has shown that the lack of digital OHS information is a significant factor contributing to the poor performance of OHS management in construction. BIM applied to OSH management has not yet yielded the same benefits as other BIM applications, such as in architectural or structural design, or project management. However, scientific literature indicates that BIM has the potential to optimize OSH management and that the construction industry, especially major projects and larger general contractors, are starting to adopt these kinds of technologies for use in OSH management. BIM has a great potential for the planning of OSH and its use at the early stages of the project has been linked to an improvement in safety conditions. Standardization of BIM for OSH is rapidly progressing. Digital4OSH is preparing a novel solution in the form of a BIM4OSH Observatory designed to enable the observation and monitoring of implementation, trends and dynamics of BIM implementation for OSH and to enable the exchange of lessons learned about BIM for OSH. The adoption of BIM paves the way for a paradigm shift in OSH management, providing stronger links between production and safety. Therefore, any initiative that improves the take up of BIM for OSH in order to reduce accidents and deaths will be very important and considered welcomed by the wider construction community

BIM4OHS observatory: central repository to monitor the status of BIM implementation for OSH: purposed architecture

DOI livro: 10.24840/978-972-752-309-2Key Technological Developments (KTDs), in recent years, have led to a step change in dealing with Occupational Safety and Health (OSH) risk management. Building Information Modelling (BIM), part of a wider trend of applying digital technology in the Architecture, Engineering, Construction and Operation (AECO) sector, has the potential to optimize the management of risks and costs of accidents at work and occupational diseases. Understanding the way OSH management can be improved using BIM is important as new processes and standards need to be created and existing procedures adapted. Currently there is no centralized sharing mechanism where countries, companies or projects can share lessons learned to help their implementation. Furthermore, there is no formal mechanism to observe and monitor trends and dynamics in the use of BIM for OSH at National, European or industry levels. Digital4OSH is a research group comprised of multidisciplinary academics and industry partners whose aim is to encourage the use of KTDs to improve OSH outcomes. Following a pilot study carried in a complex infrastructure megaproject in UK, this group proposes the development of an Observatory to overcome these gaps. The Observatory would be built on a web-based platform that can be used to obtain statistical longitudinal OSH data and provide information about the progress of national and European implementation of BIM for OSH (through dashboards); to capture, centralize and share (through factsheets) lessons learned from previous projects; to create a repository of technical and scientific information

Peaks above the Harrison-Zel'dovich spectrum due to the Quark-Gluon to Hadron Transition

The quark-gluon to hadron transition affects the evolution of cosmological perturbations. If the phase transition is first order, the sound speed vanishes during the transition, and density perturbations fall freely. This distorts the primordial Harrison-Zel'dovich spectrum of density fluctuations below the Hubble scale at the transition. Peaks are produced, which grow at most linearly in wavenumber, both for the hadron-photon-lepton fluid and for cold dark matter. For cold dark matter which is kinetically decoupled well before the QCD transition clumps of masses below $10^{-10} M_\odot$ are produced.Comment: Extended version, including evolution of density perturbations for a bag model and for a lattice QCD fit (3 new figures). Spectrum for bag model (old figure) is available in astro-ph/9611186. 9 pages RevTeX, uses epsf.sty, 3 PS figure

Temporal regulation of expression of immediate early and second phase transcripts by endothelin-1 in cardiomyocytes

Background: Endothelin-1 stimulates Gq protein-coupled receptors to promote proliferation in dividing cells or hypertrophy in terminally differentiated cardiomyocytes. In cardiomyocytes, endothelin-1 rapidly (within minutes) stimulates protein kinase signaling, including extracellular-signal regulated kinases 1/2 (ERK1/2; though not ERK5), with phenotypic/physiological changes developing from approximately 12 h. Hypertrophy is associated with changes in mRNA/protein expression, presumably consequent to protein kinase signaling, but the connections between early, transient signaling events and developed hypertrophy are unknown. Results: Using microarrays, we defined the early transcriptional responses of neonatal rat cardiomyocytes to endothelin-1 over 4 h, differentiating between immediate early gene (IEG) and second phase RNAs with cycloheximide. IEGs exhibited differential temporal and transient regulation, with expression of second phase RNAs within 1 h. Of transcripts upregulated at 30 minutes encoding established proteins, 28 were inhibited >50% by U0126 (which inhibits ERK1/2/5 signaling), with 9 inhibited 25-50%. Expression of only four transcripts was not inhibited. At 1 h, most RNAs (approximately 67%) were equally changed in total and polysomal RNA with approximately 17% of transcripts increased to a greater extent in polysomes. Thus, changes in expression of most protein-coding RNAs should be reflected in protein synthesis. However, approximately 16% of transcripts were essentially excluded from the polysomes, including some protein-coding mRNAs, presumably inefficiently translated. Conclusion: The phasic, temporal regulation of early transcriptional responses induced by endothelin-1 in cardiomyocytes indicates that, even in terminally differentiated cells, signals are propagated beyond the primary signaling pathways through transcriptional networks leading to phenotypic changes (that is, hypertrophy). Furthermore, ERK1/2 signaling plays a major role in this response

Mediators of mineralocorticoid receptor-induced profibrotic inflammatory responses in the heart

A B S T R A C T Coronary, vascular and perivascular inflammation in rats following MR (mineralocorticoid receptor) activation plus salt are well-characterized precursors for the appearance of cardiac fibrosis. Endogenous corticosterone, in the presence of the 11βHSD2 (11β hydroxysteroid dehydrogenase type 2) inhibitor CBX (carbenoxolone) plus salt, produces similar inflammatory responses and tissue remodelling via activation of MR. MR-mediated oxidative stress has previously been suggested to account for these responses. In the present study we thus postulated that when 11βHSD2 is inhibited, endogenous corticosterone bound to unprotected MR in the vessel wall may similarly increase early biomarkers of oxidative stress. Uninephrectomized rats received either DOC (deoxycorticosterone), CBX or CBX plus the MR antagonist EPL (eplerenone) together with 0.9 % saline to drink for 4, 8 or 16 days. Uninephrectomized rats maintained on 0.9 % saline for 8 days served as controls. After 4 days, both DOC and CBX increased both macrophage infiltration and mRNA expression of the p22 phox subunit of NADPH oxidase, whereas CBX, but not DOC, increased expression of the NOX2 (gp91 phox ) subunit. eNOS [endothelial NOS (NO synthase)] mRNA expression significantly decreased from 4 days for both treatments, and iNOS (inducible NOS) mRNA levels increased after 16 days of DOC or CBX; co-administration of EPL inhibited all responses to CBX. The responses characterized over this time course occurred before measurable increases in cardiac hypertrophy or fibrosis. The findings of the present study support the hypothesis that endogenous corticosterone in the presence of CBX can activate vascular MR to produce both inflammatory and oxidative tissue responses well before the onset of fibrosis, that the two MR ligands induce differential but overlapping patterns of gene expression, and that elevation of NOX2 subunit levels does not appear necessary for full expression of MR-mediated inflammatory and fibrogenic responses

Lag Length Selection for Unit Root Tests in the Presence of Nonstationary Volatility

A number of recent papers have focused on the problem of testing for a unit root in the case where the driving shocks may be unconditionally heteroskedastic. These papers have, however, taken the lag length in the unit root test regression to be a deterministic function of the sample size, rather than data-determined, the latter being standard empirical practice. We investigate the finite sample impact of unconditional heteroskedasticity on conventional data-dependent lag selection methods in augmented Dickey–Fuller type regressions and propose new lag selection criteria which allow for unconditional heteroskedasticity. Standard lag selection methods are shown to have a tendency to over-fit the lag order under heteroskedasticity, resulting in significant power losses in the (wild bootstrap implementation of the) augmented Dickey–Fuller tests under the alternative. The proposed new lag selection criteria are shown to avoid this problem yet deliver unit root tests with almost identical finite sample properties as the corresponding tests based on conventional lag selection when the shocks are homoskedastic

Aromatase Is a Direct Target of FOXL2: C134W in Granulosa Cell Tumors via a Single Highly Conserved Binding Site in the Ovarian Specific Promoter

BACKGROUND: Granulosa cell tumors (GCT) of the ovary often express aromatase and synthesize estrogen, which in turn may influence their progression. Recently a specific point mutation (C134W) in the FOXL2 protein was identified in >94% of adult-type GCT and it is likely to contribute to their development. A number of genes are known to be regulated by FOXL2, including aromatase/CYP19A1, but it is unclear which are direct targets and whether the C134W mutation alters their regulation. Recently, it has been reported that FOXL2 forms a complex with steroidogenic factor 1 (SF-1) which is a known regulator of aromatase in granulosa cells. METHODOLOGY/PRINCIPAL FINDINGS: In this work, the human GCT-derived cell lines KGN and COV434 were heterozygous and wildtype for the FOXL2:C134W mutation, respectively. KGN had abundant FOXL2 mRNA expression but it was not expressed in COV434. Expression of exogenous FOXL2:C134W in COV434 cells induced higher expression of a luciferase reporter for the ovarian specific aromatase promoter, promoter II (PII) (-516bp) than expression of wildtype FOXL2, but did not alter induction of a similar reporter for the steroidogenic acute regulatory protein (StAR) promoter (-1300bp). Co-immunoprecipitation confirmed that FOXL2 bound SF-1 and that it also bound its homologue, liver receptor homologue 1 (LRH-1), however, the C134W mutation did not alter these interactions or induce a selective binding of the proteins. A highly conserved putative binding site for FOXL2 was identified in PII. FOXL2 was demonstrated to bind the site by electrophoretic mobility shift assays (EMSA) and site-directed mutagenesis of this element blocked its differential induction by wildtype FOXL2 and FOXL2:C134W. CONCLUSIONS/SIGNIFICANCE: These findings suggest that aromatase is a direct target of FOXL2:C134W in adult-type GCT via a single distinctive and highly conserved binding site in PII and therefore provide insight into the pathogenic mechanism of this mutation