Cardiac Function and Architecture Are Maintained in a Model of Cardiorestricted Overexpression of the Prorenin-Renin Receptor

The (pro)renin-renin receptor, (P)RR has been claimed to be a novel element of the renin-angiotensin system (RAS). The function of (P)RR has been widely studied in renal and vascular pathology but the cardio-specific function of (P)RR has not been studied in detail. We therefore generated a transgenic mouse (Tg) with cardio-restricted (P)RR overexpression driven by the alpha-MHC promotor. The mRNA expression of (P)RR was ∼170-fold higher (P<0.001) and protein expression ∼5-fold higher (P<0.001) in hearts of Tg mice as compared to non-transgenic (wild type, Wt) littermates. This level of overexpression was not associated with spontaneous cardiac morphological or functional abnormalities in Tg mice. To assess whether (P)RR could play a role in cardiac hypertrophy, we infused ISO for 28 days, but this caused an equal degree of cardiac hypertrophy and fibrosis in Wt and Tg mice. In addition, ischemia-reperfusion injury was performed in Langendorff perfused isolated mouse hearts. We did not observe differences in parameters of cardiac function or damage between Wt and Tg mouse hearts under these conditions. Finally, we explored whether the hypoxia sensing response would be modulated by (P)RR using HeLa cells with and without (P)RR overexpression. We did not establish any effect of (P)RR on expression of genes associated with the hypoxic response. These results demonstrate that cardio-specific overexpression of (P)RR does not provoke phenotypical differences in the heart, and does not affect the hearts’ response to stress and injury. It is concluded that increased myocardial (P)RR expression is unlikely to have a major role in pathological cardiac remodeling

Efficient Use of Water in the Textile Finishing Industry

Identification and exploitation of non-conventional water sources is a priority for many industrial sectors, especially for the textile finishing industry. Therefore a multicriteria integrated and coherent methodology to support the implementation of sustainable water reuse is essential. The methodology conceived within the EU project TOWEF0 (TOWards EFfluent zero) is presented in this paper, together with the tools to carry out all the steps required by its application. A process data collection for technical/economical evaluation in textile companies was performed and integrated with a characterisation of the process effluents in terms of treatability and reusability. Feasibility evaluations of effluents treatment for reuse were performed and reuse tests in textile processes were carried out. These information allowed for the design of optimised water reuse schemes by Water Pinch application, whereas the Life Cycle Assessment (LCA) permitted the evaluation and comparison of water reuse scenarios. Physical-chemical and eco-toxicological monitoring campaigns of textile discharges and receiving water bodies were conducted as well. All these specific results, valuable and applicable independently, when integrated into the TOWEF0 methodology, generate viable solutions in accordance with the fundamentals of the IPPC DirectiveJRC.H.5-Rural, water and ecosystem resource

Assessment of ex vivo ischemia reperfusion injury by Langendorff isolated heart perfusion in (P)RR Tg mice.

<p>A) Leakage of lactate dehydrogenase (LDH) before and after ischemia and 5, 10 and 30 min after reperfusion of Wt and Tg mice. No significant difference was found between Wt and Tg mice. B) Leakage of creatine kinase (CK) before and after ischemia and 5, 10 and 30 min after reperfusion of Wt and Tg mice. No significant difference was found between Wt and Tg mice. C) Functional recovery; LV developed pressure (LVDP) for Wt and (P)RR Tg hearts after 30 min of reperfusion. D) Rate–pressure product (RPP) for Wt and (P)RR Tg hearts after 30 min of reperfusion. Values are means ± SEM; n = 6 per group.</p

Western blot of ERK1/2, p-38 and HSP47.

<p><b>A)</b> Western blot of the expression of ERK1/2, p-38 and HSP47 proteins from hearts of Tg and Wt mice with and without ISO treatment. B) Quantification of the ratio of phospho−/total Erk1/2 from hearts of Tg and Wt mice with and without ISO treatment. C) Quantification of the ratio of phospho−/total p-38 from hearts of Tg and Wt mice with and without ISO treatment. D) Quantification of the HSP47 protein expression from hearts of Tg and Wt mice with and without ISO treatment. * <i>P<0.05</i>, Wt vs. Tg.</p

Assessment of isoproterenol induced cardiac hypertrophy in (P)RR transgenic mice.

<p>A) Left-ventricular weight (LV-W) to tibia length (TL)in adult Wt and Tg mice subjected to saline or ISO infusion for 28 days (N = 7 mice in each group). B) <i>ANP</i> mRNA expression (normalized to <i>36B4</i>) in adult Wt and Tg mice subjected to saline or ISO infusion for 28 days (N = 7 mice in each group). C) Renin mRNA expression (normalized to <i>36B4</i>) in adult Wt and Tg mice subjected to saline or ISO infusion for 28 days (N = 7 mice in each group). D) <i>α-MHC/β-MHC</i> (ratio) mRNA expression (normalized to <i>36B4</i>) in adult Wt and Tg mice subjected to saline or ISO infusion for 28 days (N = 7 mice in each group). E and F) Masson’s trichrome staining (bar size 100 µm) to assess myocardial fibrosis in adult Wt and Tg mice subjected to saline (sham procedure) or ISO infusion for 28 days; E: quantification, and F: typical examples of Masson’s trichrome staining. G and H) FITC-WGA staining to assess myocyte hypertrophy in adult Wt and Tg mice subjected to saline (sham procedure) or ISO infusion for 28 days; G: quantification, and H: typical examples of FITC-WGA staining. *<i>P</i><0.05, **<i>P</i><0.01, sham vs. ISO between Wt mice; ^#<i>P</i><0.05, ^##<i>P</i><0.01, sham vs. ISO between Tg mice and ^§<i>P</i><0.05, ISO treatment between Wt and Tg mice.</p

Effect of (P)RR overexpression on stress related gene expression due to hypoxia.

<p>Gene expression levels in HeLa cells under normoxia, DFO (deferoxamine) treatment and hypoxia; HeLa cells without (fine bars) and with (black bars) (P)RR overexpression. A) <i>ADM</i> B) <i>VEGF</i> C) <i>c-jun</i> D) <i>c-fos</i> (all normalized to <i>GAPDH</i>). *<i>P<0.05</i>, ***<i>P<0.001</i>, normoxia vs. DFO or normoxia vs. hypoxia, under control condition. ^#<i>P<0.05, </i>^##<i>P<0.01</i> and ^###<i>P<0.001</i>, normoxia vs. DFO or normoxia vs. hypoxia, under (P)RR overexpression.</p