Targeting a phospho-STAT3-miRNAs pathway improves vesicular hepatic steatosis in an in vitro and in vivo model

Non-alcoholic fatty liver disease (NAFLD) is a leading cause of chronic liver disease. Although genetic predisposition and epigenetic factors contribute to the development of NAFLD, our understanding of the molecular mechanism involved in the pathogenesis of the disease is still emerging. Here we investigated a possible role of a microRNAs-STAT3 pathway in the induction of hepatic steatosis. Differentiated HepaRG cells treated with the fatty acid sodium oleate (fatty dHepaRG) recapitulated features of liver vesicular steatosis and activated a cell-autonomous inflammatory response, inducing STAT3-Tyrosine-phosphorylation. With a genome-wide approach (Chromatin Immunoprecipitation Sequencing), many phospho-STAT3 binding sites were identified in fatty dHepaRG cells and several STAT3 and/or NAFLD-regulated microRNAs showed increased expression levels, including miR-21. Innovative CARS (Coherent Anti-Stokes Raman Scattering) microscopy revealed that chemical inhibition of STAT3 activity decreased lipid accumulation and deregulated STAT3-responsive microRNAs, including miR-21, in lipid overloaded dHepaRG cells. We were able to show in vivo that reducing phospho-STAT3-miR-21 levels in C57/BL6 mice liver, by long-term treatment with metformin, protected mice from aging-dependent hepatic vesicular steatosis. Our results identified a microRNAs-phosphoSTAT3 pathway involved in the development of hepatic steatosis, which may represent a molecular marker for both diagnosis and therapeutic targeting

Genome-wide identification of direct HBx genomic targets

Background: The Hepatitis B Virus (HBV) HBx regulatory protein is required for HBV replication and involved in HBV-related carcinogenesis. HBx interacts with chromatin modifying enzymes and transcription factors to modulate histone post-translational modifications and to regulate viral cccDNA transcription and cellular gene expression. Aiming to identify genes and non-coding RNAs (ncRNAs) directly targeted by HBx, we performed a chromatin immunoprecipitation sequencing (ChIP-Seq) to analyse HBV recruitment on host cell chromatin in cells replicating HBV. Results: ChIP-Seq high throughput sequencing of HBx-bound fragments was used to obtain a high-resolution, unbiased, mapping of HBx binding sites across the genome in HBV replicating cells. Protein-coding genes and ncRNAs involved in cell metabolism, chromatin dynamics and cancer were enriched among HBx targets together with genes/ncRNAs known to modulate HBV replication. The direct transcriptional activation of genes/miRNAs that potentiate endocytosis (Ras-related in brain (RAB) GTPase family) and autophagy (autophagy related (ATG) genes, beclin-1, miR-33a) and the transcriptional repression of microRNAs (miR-138, miR-224, miR-576, miR-596) that directly target the HBV pgRNA and would inhibit HBV replication, contribute to HBx-mediated increase of HBV replication. Conclusions: Our ChIP-Seq analysis of HBx genome wide chromatin recruitment defined the repertoire of genes and ncRNAs directly targeted by HBx and led to the identification of new mechanisms by which HBx positively regulates cccDNA transcription and HBV replication

Drivers of success in implementing sustainable tourism policies in urban areas

The existing literature in the field of sustainable tourism highlights a number of barriers that impede the implementation of policies in this area. Yet, not many studies have so far considered the factors that would contribute to putting this concept into practice, and few address the case of urban areas. The concept of sustainability has only received limited attention in urban tourism research, even though large cities are recognised as one of the most important tourist destinations that attract vast numbers of visitors. Adopting a case study approach, this paper discusses a number of drivers of success identified by policy-makers in London to contribute to the implementation of sustainable tourisms policies at the local level, and briefly looks at the relationship between these drivers and the constraints perceived by the respondents to hinder the implementation of such policies in practice. These findings may help policy-makers in other large cities to successfully develop and implement policies towards sustainable development of tourism in their area

Students’ Confidence and Interest in Palliative and Bereavement Care: A European Study

As part of a European Erasmus Plus project entitled Death Education for Palliative Psychology, this study assessed the ways in which Master’s Degree students in psychology and the creative arts therapies self-rated their confidence and interest in death education and palliative and bereavement care. In five countries (Austria, Israel, Italy, Poland, Romania), 344 students completed an online questionnaire, and 37 students were interviewed to better understand their views, interest, and confidence. The results revealed some significant differences between countries, and showed that older respondents with previous experience as formal caregivers for end-of-life clients showed greater interest in obtaining practical clinical competence in these fields. A mediation analysis indicated that students’ previous care experiences and past loss experiences were related to students’ current interest in death education and palliative and bereavement care through the mediation of their sense of confidence in this field. The qualitative findings identified five shared themes: life and death, learning about death, the psychological burden, personal experience and robust training, and four key training needs. Overall, students’ interest in studying and working with terminal illness and death are rooted in internal resources, a preliminary sense of confidence, but also external requirements

Italian Ragweed Pollen Inventory

This study provides the first spatially detailed and complete inventory of Ambrosia pollen sources in Italy – the third largest centre of ragweed in Europe. The inventory relies on a well tested top-down approach that combines local knowledge, detailed land cover, pollen observations and a digital elevation model that assumes permanent ragweed populations mainly grow below 745m. The pollen data were obtained from 92 volumetric pollen traps located throughout Italy during 2004-2013. Land cover is derived from Corine Land cover information with 100m resolution. The digital elevation model is based on the NASA shuttle radar mission with 90m resolution. The inventory is produced using a combination of ArcGIS and Python for automation and validated using cross-correlation and has a final resolution of 5km x 5km. The method includes a harmonization of the inventory with other European inventories for the Pannonian Plain, France and Austria in order to provide a coherent picture of all major ragweed sources. The results show that the mean annual pollen index varies from 0 in South Italy to 6779 in the Po Valley. The results also show that very large pollen indexes are observed in the Milan region, but this region has smaller amounts of ragweed habitats compared to other parts of the Po Valley and known ragweed areas in France and the Pannonian Plain. A significant decrease in Ambrosia pollen concentrations was recorded in 2013 by pollen monitoring stations located in the Po Valley, particularly in the Northwest of Milan. This was the same year as the appearance of the Ophraella communa leaf beetle in Northern Italy. These results suggest that ragweed habitats near to the Milan region have very high densities of Ambrosia plants compared to other known ragweed habitats in Europe. The Milan region therefore appears to contain habitats with the largest ragweed infestation in Europe, but a smaller amount of habitats is a likely cause the pollen index to be lower compared to central parts of the Pannonian Plain. A low number of densely packed habitats may have increased the impact of the Ophraella beetle and might account for the documented decrease in airborne Ambrosia pollen levels, an event that cannot be explained by meteorology alone. Further investigations that model atmospheric pollen before and after the appearance of the beetle in this part of Northern Italy are needed to assess the influence of the beetle on airborne Ambrosia pollen concentrations. Future work will focus on short distance transport episodes for stations located in the Po Valley, and long distance transport events for stations in Central Italy that exhibit peaks in daily airborne Ambrosia pollen levels

Prognostic impact of genetic characterization in the GIMEMA LAM99P multicenter study for newly diagnosed acute myeloid leukemia

Recent advances in genetic characterization of acute myeloid leukemia indicate that combined cytogenetic and molecular analyses provide better definition of prognostic groups. The aim of this study was to verify this prospectively in a large group of patients