Functional modulation of the transient outward current Ito by KCNE beta-subunits and regional distribution in human non-failing and failing hearts

Objectives: The function of Kv4.3 (KCND3) channels, which underlie the transient outward current I,, in human heart, can be modulated by several accessory subunits such as KChIP2 and KCNE1-KCNE5. Here we aimed to determine the regional expression of Kv4.3, KChIP2, and KCNE mRNAs in non-failing and failing human hearts and to investigate the functional consequences of subunit coexpression in heterologous expression systems. Methods: We quantified mRNA levels for two Kv4.3 isoforms, Kv4.3-S and Kv4.3-L, and for KChIP2 as well as KCNE1-KCNE5 with real-time RT-PCR. We also studied the effects of KCNEs on Kv4.3 + KChIP2 current characteristics in CHO cells with the whole-cell voltage-clamp method. Results: In non-failing hearts, low expression was found for KCNE1, KCNE3, and KCNE5, three times higher expression for KCNE2, and 60 times higher for KCNE4. Transmural gradients were detected only for KChIP2 in left and right ventricles. Compared to non-failing tissue, failing hearts showed higher expression of Kv4.3-L and KCNE1 and lower of Kv4.3-S, KChIP2, KCNE4, and KCNE5. In CHO cells, Kv4.3 + KChIP2 currents were differentially modified by co-expressed KCNEs: time constants of inactivation were shorter with KCNE1 and KCNE3-5 while time-to-peak was decreased, and V-0.5 of steady-state inactivation was shifted to more negative potentials by all KCNE subunits. Importantly, KCNE2 induced a unique and prominent 'overshoot' of peak current during recovery from inactivation similar to that described for human I-to while other KCNE subunits induced little (KCNE4,5) or no overshoot. Conclusions: All KCNEs are expressed in the human heart at the transcript level. Compared to It. in native human myocytes, none of the combination of KChIP2 and KCNE produced an ideal congruency in current characteristics, suggesting that additional factors contribute to the regulation of the native I-to channel

METRO - The role and future perspectives of Cohesion Policy in the planning of Metropolitan Areas and Cities. Annex III: Metropolitan City of Turin case study

The Metropolitan City of Turin, which is one of the case studies of the ESPON METRO project, is a wide area, second level local authority which replaced the former Province of Turin from January 1st, 2015. The metropolitan institution brings together 312 municipalities, covering a very large and heterogeneous territory, from dense urban areas to small towns and villages, much larger than the functional urban area of Turin. It is the largest metropolitan city in Italy, fourth in population size (2.2Mln inhabitants) and seventh in population density (330 inh/km2). Metropolitan Cities are administrative units formally established in Italy by the reform of local authorities (National Law 56/2014), replacing the respective Province authorities. The Italian Metropolitan cities still perform all the functions of the previous Province authorities, and have additional functions, such as strategic, spatial and mobility planning, organization of coordinated systems for the management of public services, mobility and transport, promotion and coordination of digitalization and economic and social development. Despite the high level of institutionalization and competences of the Metropolitan City of Turin, which is formally acknowledged as an entity also enjoying a supranational relevance when it comes to access EU funds, the institution does not have relevant role and competences in the elaboration of key policy and programming documents of the EU cohesion policy and in their management and implementation, while the Region and the national level are the main actors. In this case study report, the potential and actual role of the Metropolitan City of Turin in the implementation of the cohesion policy, and the impact of the cohesion policy on metropolitan development and governance are analysed. After a territorial and socio-economic contextualization of the area, the metropolitan governance and cooperation activities are presented. Then, the cohesion policy institutional architecture and policy instruments are presented, and the role of the metropolitan actors in the planning and implementation of the cohesion policy is examined, as well as its coordination with metropolitan governance. The cohesion policy impact on metropolitan development and governance is then presented, analyzing the coherence and synergies of its instruments and goals with metropolitan ones, the funding magnitude and the related impact. Issues related to the response to the COVID-19 emergency are presented in each of the three core sections of the report. Finally, the last section of the report summarizes the main challenges and recommendations

ESPON SUPER – Sustainable Urbanisation and land-use Practices in European Regions. A GUIDE TO SUSTAINABLE URBANISATION AND LAND-USE

Guides help you do things. You turn to them when you need to find out how to solve a problem. They are a form of knowledge transfer, written by experts in a way that is accessible and helpful to a wide audience. This guide was written by the researchers engaged in the ESPON 2020 applied research project on Sustainable Urbanisation and Land-Use Practices in European Regions (SUPER). It aims to help people and institutions engaged with land-use management at various levels across Europe to promote sustainable urbanisation in their territories. Overall, the guide offers information, ideas and perspectives to help decision-makers and policymakers to proactively contribute to more equal, balanced, and sustainable territorial development. The decision to convert land to a different use influences our quality of life and that of future generations, and, as this Guide shows, a large toolbox of interventions exists that can help alter prevailing land-use practices. Choosing among them is a tough decision, and implementation may require strong political commitment and bold leadership. We hope that this Guide provides the inspiration to accept this challenge

Angiogenic potential in biological hydrogels

Hydrogels are three-dimensional (3D) materials able to absorb and retain water in large amounts while maintaining their structural stability. Due to their considerable biocompatibility and similarity with the body\u2019s tissues, hydrogels are one of the most promising groups of biomaterials. The main application of these hydrogels is in regenerative medicine, in which they allow the formation of an environment suitable for cell differentiation and growth. Deriving from these hydrogels, it is, therefore, possible to obtain bioactive materials that can regenerate tissues. Because vessels guarantee the right amount of oxygen and nutrients but also assure the elimination of waste products, angiogenesis is one of the processes at the base of the regeneration of a tissue. On the other hand, it is a very complex mechanism and the parameters to consider are several. Indeed, the factors and the cells involved in this process are numerous and, for this reason, it has been a challenge to recreate a biomaterial able to adequately sustain the angiogenic process. However, in this review the focal point is the application of natural hydrogels in angiogenesis enhancing and their potential to guide this process

The correlation between low back pain and strength training in elite athletes: a literature review

This study aims to analyze the correlation between LBP and elite athletes who practice sports where strength training intervenes via weightlifting. To analyse the correlation between low back pain and athletes who practice sports where strength training programs, a narrative review was conducted by two independent author through MEDLINE database search. Inclued study’s methodology quality has been evaluated using NIH quality assessment tool for Observational Cohort and Cross-Sectional Studies. Out of 830 retrieved articles, after titles, abstracts and full text assessment, four studies met the inclusion criteria and were included in the present narrative review. The NIH total score ranged from 10 to 12 points. Demographic and sport-specific factors can influence the prevalence of LBP. Our findings highlight the importance of developing future research to provide prevention programs to reduce the incidence of LBP, taking into account the demographics of athletes and the unique nature of their sport activity

High-throughput assessment of the antibody profile in ovarian cancer ascitic fluids

The identification of effective biomarkers for early diagnosis, prognosis, and response to treatments remains a challenge in ovarian cancer (OC) research. Here, we present an unbiased high-throughput approach to profile ascitic fluid autoantibodies in order to obtain a tumor-specific antigen signature in OC. We first reported the reactivity of immunoglobulins (Igs) purified from OC patient ascites towards two different OC cell lines. Using a discovery set of Igs, we selected tumor-specific antigens from a phage display cDNA library. After biopanning, 700 proteins were expressed as fusion protein and used in protein array to enable large-scale immunoscreening with independent sets of cancer and noncancerous control. Finally, the selected antigens were validated by ELISA. The initial screening identified eight antigenic clones: CREB3, MRPL46, EXOSC10, BCOR, HMGN2, HIP1R, OLFM4, and KIAA1755. These antigens were all validated by ELISA in a study involving ascitic Igs from 153 patients (69 with OC, 34 with other cancers and 50 without cancer), with CREB3 showing the highest sensitivity (86.95%) and specificity (98%). Notably, we were able to identify an association between the tumor-associated (TA) antibody response and the response to a first-line tumor treatment (platinum-based chemotherapy). A stronger association was found by combining three antigens (BCOR, CREB3, and MRLP46) as a single antibody signature. Measurement of an ascitic fluid antibody response to multiple TA antigens may aid in the identification of new prognostic signatures in OC patients and shift attention to new potentially relevant targets

The RNA-binding protein ILF3 binds to transposable element sequences in SINEUP lncRNAs

Transposable elements (TEs) compose about half of the mammalian genome and, as embedded sequences, up to 40% of long noncoding RNA (lncRNA) transcripts. Embedded TEs may represent functional domains within lncRNAs, providing a structured RNA platform for protein interaction. Here we show the interactome profile of the mouse inverted short interspersed nuclear element (SINE) of subfamily B2 (invSINEB2) alone and embedded in antisense (AS) ubiquitin C-terminal hydrolase L1 (Uchl1), an lncRNA that is AS to Uchl1 gene. AS Uchl1 is the representative member of a functional class of AS lncRNAs, named SINEUPs, in which the invSINEB2 acts as effector domain (ED)-enhancing translation of sense protein-coding mRNAs. By using RNA-interacting domainome technology, we identify the IL enhancer-binding factor 3 (ILF3) as a protein partner of AS Uchl1 RNA. We determine that this interaction is mediated by the RNA-binding motif 2 of ILF3 and the invSINEB2. Furthermore, we show that ILF3 is able to bind a free right Arthrobacter luteus (Alu) monomer sequence, the embedded TE acting as ED in human SINEUPs. Bioinformatic analysis of Encyclopedia of DNA Elements-enhanced cross-linking immunoprecipitation data reveals that ILF3 binds transcribed human SINE sequences at transcriptome-wide levels. We then demonstrate that the embedded TEs modulate AS Uchl1 RNA nuclear localization to an extent moderately influenced by ILF3. This work unveils the existence of a specific interaction between embedded TEs and an RNA-binding protein, strengthening the model of TEs as functional modules in lncRNAs.-Fasolo, F., Patrucco, L., Volpe, M., Bon, C., Peano, C., Mignone, F., Carninci, P., Persichetti, F., Santoro, C., Zucchelli, S., Sblattero, D., Sanges, R., Cotella, D., Gustincich, S. The RNA-binding protein ILF3 binds to transposable element sequences in SINEUP lncRNAs