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Nell’ambito della sua mission di Agenzia Nazionale per le nuove tecnologie, l’energia e lo sviluppo economico sostenibile, l’ENEA è focalizzata in modo preminente sulla ricerca ‘applicata’, ovvero sul rendere disponibili tecnologie innovative e servizi avanzati nei diversi settori di competenza, al mondo delle imprese, alle pubbliche amministrazioni e agli stessi cittadini . Da questo punto di vista, l’economia circolare è una delle priorità strategiche e si concretizza attraverso numerosi progetti operativi con l’obiettivo di andare ‘oltre gli slogan’ e tradurre in realtà il principio fondante della chiusura dei cicli, quale volano di crescita e competitività e, allo stesso tempo, strumento essenziale per un modello di società incentrato sulla sostenibilità. In questo opuscolo sono illustrati alcuni dei progetti, delle iniziative e delle attività, ma anche gli strumenti, le metodologie, gli approcci e i modelli che ENEA sta mettendo in campo per supportare la trasformazione verso l’economia circolare, con un focus anche sulla formazione ed informazione, aspetti di grande rilievo per operare il cambiamento culturale e di comportamenti necessario. In questi progetti il Dipartimento “Sostenibilità dei Sistemi Produttivi e Territoriali” (SSPT) ha un ruolo di primo piano, come coordinatore o referente di attività in collaborazione con altri Dipartimenti dell’Agenzia, partner nazionali ed internazionali, imprese e istituzioni centrali locali, ed altri soggetti a livello territoriale e urbano. Queste attività impegnano oltre cento ricercatori e tecnologi del Dipartimento SSPT e una rete di infrastrutture, hall tecnologiche, impianti pilota e laboratori analitici avanzati specializzati nell’eco-innovazione di processo, di prodotto e di sistema. I progetti consentono di applicare l’economia circolare in diversi ambiti applicativi quali: • Aree urbane: ENEA progetta e sviluppa modelli e sistemi di gestione integrata e circolare di funzionamento urbano per città più sostenibili, circolari e inclusive, per promuovere stili di vita e consumo sostenibili, per effettuare una corretta gestione dell’acqua e dei rifiuti urbani anche in chiave di valorizzazione, salvaguardare salute e sicurezza, stimolare le industrie culturali, turismo e best practices in contesti urbani e periurbani; • Territorio e mare: ENEA pianifica e sviluppa processi e metodologie per la gestione sostenibile di territorio e mare in ottica di economia circolare attraverso la Gestione porti e aree costiere, il Turismo sostenibile, lo sviluppo della filiera ittica; • Sistema industriale: ENEA sviluppa e implementa tecnologie e metodologie per modelli di produzione e consumo più sostenibili e rigenerativi a supporto dell’industria con tecnologie innovative e nuovi modelli di business (simbiosi industriale, Modelli di circular design, Sharing economy, etc), strumenti per le imprese, riqualificazione di siti industriali in ottica circolare; • Catena del valore: ENEA sviluppa approcci di sistema per promuovere e facilitare la chiusura dei cicli nelle filiere produttive e lungo il ciclo di vita di prodotti e materiali attraverso attività per promuovere la collaborazione tra diversi attori e settori, approccio integrato e multidisciplinare (life cycle thinking e misura della circolarità), nuovi modelli di business e analisi di mercato.

In the wake of the 2008 financial crisis the green economy emerged as a policy approach to help solve two problems: the economic slowdown and consequent loss of jobs, and the continuing deterioration of the natural environment, most visible in terms of climate change and ecosystem degradation. This potential double dividend led many European countries to adopt recovery policies in 2008 and 2009 that had a green economy focus. However, as policy focus shifted to the fiscal consolidation and sovereign debt crises in Europe, the concept of the green economy began to lose appeal within short‑term macro‑economic policies. At the same time, the green economy became a pillar of major European and international strategies: most notably within the Europe 2020 strategy adopted in 2010 by the EU to drive sustainable growth, and in the Rio+20 outcome The future we want (UN, 2012) as a tool for achieving sustainable development. The green economy can now be seen as an approach that can achieve a structural and permanent transformation of the economy. This report is guided by the definition of a green economy as: …. combining enhanced resource efficiency with environmental resilience, while boosting prosperity and equity in society (4). Specifically, it analyses macro‑level trends in Europe since the 1990s and addresses those economic factors, drivers, and outcomes that could support a stable shift to a resource‑efficient green economy (hereafter called simply green economy): green innovation, green finance, environmental fiscal reforms and the changing sectoral mix of the economy. A specific focus is the link between the shift to a green economy and the EU policy framework. As illustrated in Towards a green economy in Europe — EU environmental policy targets and objectives 2010–2050 (EEA, 2013), a wide range of specific policies is already in place to support change. However, this report also considers the links between the green economy and policies outside the environmental sector, such as manufacturing and competitiveness policies. The logic of the report's approach and structure is presented in the figure on the following page. Chapters 1–4 highlight the macro‑level trends towards a green economy emerging from the policy framework and the major transformation of the EU economy over recent decades. In Chapter 1, different concepts and definitions of the green economy are discussed. In addition, a brief history of the emergence of the concept of a green economy in the wake of the financial crisis is given. The EU policy landscape, focusing on different EU policy initiatives that promote the green economy, is addressed in Chapter 2. In Chapter 3, recent developments and progress in achieving environmental policy targets are studied. The changing structure of the EU's economy in terms of environmental pressures, and the extent to which manufacturing, services, and trade can play a role, are analysed in Chapter 4. Given the combination of policy pushes and macro‑level transformations of the economy, Chapters 5–8 address key enabling factors that can help to promote or hasten the development of a green economy (Figure I.1). Chapter 5 focuses on the role of eco‑innovation diffusion as an established enabling condition in Europe. The role of the international transfer of green knowledge, another enabling factor, is presented in Chapter 6. The role of environmental fiscal reforms and economic instruments are analysed in Chapter 7. Finally, the role of finance, and particularly of innovative green finance as an enabling condition of the green economy, is analysed in Chapter 8. In short, eco‑innovation, green knowledge transfer, environmental fiscal reform and green finance (Chapters 5–8) are seen as levers for effecting a shift within the framework of favourable and unfavourable macro trends emerging from EU policies and the transformation of its economic structure (Chapters 1–4). With this logic, the report seeks to contribute to an understanding of the major forces that promote or hinder progress in Europe, and of the policies needed to guide it.

Several reports suggest that xanthine dehydrogenase (XDH) and its oxidase form (XO) play an important role in various forms of ischemic and vascular injuries. Recently we have demonstrated that 17β-estradiol (E2) induces a significant decrease of the expression and activity of XDH and its conversion to XO in human mammary epithelial cells. E2 is known to induce upregulation of eNOS gene expression in aortic endothelial cells. In light of the ability of XO-derived O2•¯ to combine with •NO to yield ONOO¯, and considering that ONOO¯ converts XDH to XO, it is important to protect tissues against the XO increased activity and ROS increased production, that would in turn react with •NO to augment ONOO¯ production, thus creating a vicious cycle of oxidative stress. Our previous studies have indicated that sildenafil has a protective effect on human mammary epithelial cells as a consequence of the inhibition of XO and of the resulting decrease of free oxygen radical that may influence the expression of NADPH oxidase and PDE-5. We report that the contemporary inhibitory effect played by sildenafil on XO and PDE-5 is due to the structural modification induced by O2•¯, which involves the release of a piperazine group able to inhibit XO.

PMID: 18615489 [PubMed - in process]The in vitro effects of polyalkyl-imide hydrogel (PAI) containing 96% pyrogen-free water on the viability, apoptosis, cell shape and metabolic activities of murine 3T3 and human Detroit 555 fibroblasts were investigated. Analysis of the viscous-elastic properties and the ultrastructure of PAI, performed by rheometer and AFM respectively, showed that the material has the typical characteristics of hydrogel, including a three-dimensional configuration of molecules arranged in a regular network with many discrete caveolae where most of the water is captured. Hydrogel biocompatibility was found to be high for both cell lines, with some differences. Cell viability decreased more in 3T3 cells than Detroit 550 fibroblasts when cultured in the presence of 100 mg/ml hydrogel but not at concentrations of 25 and 50 mg/ml hydrogel. The period of incubation with PAI (24 and 48 h) only partially affected cell viability. Apoptosis, most likely due to cells’ inability to adhere to the polymer, was the only type of cell death observed. Fibroblasts grown in the presence of polymer were always metabolically active since they continued to synthesize collagen. In conclusion, PAI hydrogel, even at high concentrations, was biocompatible for both fibroblasts, but in particular for human cells, thus encouraging its use as a dermal filler.

An important requirement in cost-efficient lignocellulosics-to-ethanol process is to achieve concentrated hydrolyzates. The research in the last years has been focused on improving the enzymes performances in terms of internal dosage of the specific components and increased thermostability. These improved features could facilitate the processing of concentrated biomass suspensions up to 20-30% solids. However, the most important problem in the hydrolysis at high solid loadings is related to the fact that, these suspensions have a strong non-Newtonian behavior with high viscosities values that cause mixing and mass transfer limitations, and, presumably increased inhibition by intermediates. In this paper preliminary results from the rheological characterization of concentrated biomass suspensions are presented with the scope of gaining data for the process design and optimization. In particular various geometries have been evaluated for the rheological characterizations of these fluids (namely vane, plate/plate, cone/plate), and several tests were done to measure the viscosity, the, the consistency and fluidity index under different process conditions.., Proceedings of the 20th European Biomass Conference and Exhibition, 18-22 June 2012, Milan, Italy, pp. 1681-1684

Hepatitis A virus infection is usually asymptomatic in children. Classic symptomatic forms and atypical clinical manifestations are known. We report a paediatric case of hepatitis A with marked cholestasis, treated with steroids, and with an unusual prolonged course.

Acute acalculous cholecystitis (CAA) is very rare in children and it is usually related to infectious agents. We report 2 paediatric cases of CAA complicating hepatitis type A, with a favourable evolution with conservative treatment.

Extracellular vesicles (EVs) constitute one of the main mechanisms by which cells communicate with the surrounding tissue or at distance. Vesicle secretion is featured by most cell types, and adult mesenchymal stromal cells (MSCs) of different tissue origins have shown the ability to produce them. In recent years, several reports disclosed the molecular composition and suggested clinical indications for EVs derived from adult MSCs. The parental cells were already known for their roles in different disease settings in regulating inflammation, immune modulation, or transdifferentiation to promote cell repopulation. Interestingly, most reports also suggested that part of the properties of parental cells were maintained by isolated EV populations. This review analyzes the recent development in the field of cell-free therapies, focusing on several adult tissues as a source of MSC-derived EVs and the available clinical data from in vivo models.

Mesenchymal stromal/stem cells (MSCs) are believed to function in vivo as a homeostatic tool that shows therapeutic properties for tissue repair/regeneration. Conventionally, these cells are expanded in two-dimensional (2D) cultures, and, in that case, MSCs undergo genotypic/phenotypic changes resulting in a loss of their therapeutic capabilities. Moreover, several clinical trials using MSCs have shown controversial results with moderate/insufficient therapeutic responses. Different priming methods were tested to improve MSC effects, and three-dimensional (3D) culturing techniques were also examined. MSC spheroids display increased therapeutic properties, and, in this context, it is crucial to understand molecular changes underlying spheroid generation. To address these limitations, we performed RNA-seq on human amnion-derived MSCs (hAMSCs) cultured in both 2D and 3D conditions and examined the transcriptome changes associated with hAMSC spheroid formation. We found a large number of 3D culture-sensitive genes and identified selected genes related to 3D hAMSC therapeutic effects. In particular, we observed that these genes can regulate proliferation/differentiation, as well as immunomodulatory and angiogenic processes. We validated RNA-seq results by qRT-PCR and methylome analysis and investigation of secreted factors. Overall, our results showed that hAMSC spheroid culture represents a promising approach to cell-based therapy that could significantly impact hAMSC application in the field of regenerative medicine.

Aiming at valorizing the ricotta cheese exhausted whey (RCEW), one of the most abundant by-products from the dairy industry, a biotechnological protocol to obtain bioactive peptides with angiotensin-I-converting enzyme (ACE)—inhibitory activity was set up. The approach was based on the combination of membrane filtration and fermentation. A Lactobacillus helveticus strain selected to be used as starter for the fermentation of the ultrafiltration protein-rich retentate (R-UF) obtained from RCEW. The fermented R-UF was characterized by a high anti-ACE activity. Peptides responsible for the bioactivity were purified and identified through nano-LC–ESI–MS/MS. The sequences identified in the purified active fractions of the fermented R-UF showed partial or complete overlapping with previously reported κ-casein antihypertensive fragments. The fermented R-UF was spray-dried and used to enrich ricotta cheese at different fortification level (1 and 5% w/w). An integrated approach including the assessment of the microbiological, chemical, functional, textural, and sensory properties was used to characterize the fortified products. A significantly higher anti-ACE activity was found in the ricotta cheese fortified with fermented R-UF as compared to the control and to the samples obtained with the unfermented R-UF fraction at the same levels of fortification. In particular, a 100 g portion of the ricotta cheese produced at 5% fortification level contained circa 30 mg of bioactive peptides. The fortification led to a moderate acidification, increased hardness and chewiness, and decreased the milk odor and taste of the ricotta cheese as compared to the control, while flavor persistence and sapidity improved.

The prevalence of end-stage liver diseases has reached very high levels globally. The election treatment for affected patients is orthotopic liver transplantation, which is a very complex procedure, and due to the limited number of suitable organ donors, considerable research is being done on alternative therapeutic options. For instance, the use of cell therapy, such as the transplantation of hepatocytes to promote liver repair/regeneration, has been explored, but standardized protocols to produce suitable human hepatocytes are still limited. On the other hand, liver progenitor and multipotent stem cells offer potential cell sources that could be used clinically. Different studies have reported regarding the therapeutic effects of transplanted mesenchymal stromal/stem cells (MSCs) on end-stage liver diseases. Moreover, it has been shown that delivery of MSC-derived conditioned medium (MSC-CM) can reduce cell death and enhance liver proliferation in fulminant hepatic failure. Therefore, it is believed that MSC-CM contains many factors that probably support liver regeneration. In our work, we used an in vitro model of human liver organoids to study if the paracrine components secreted by human amnion-derived MSCs (hAMSCs) affected liver stem/progenitor cell differentiation. In particular, we differentiated liver organoids derived from bipotent EpCAM+ human liver cells and tested the effects of hAMSC secretome, derived from both two-dimensional (2D) and three-dimensional (3D) hAMSC cultures, on that model. Our analysis showed that conditioned medium (CM) produced by 3D hAMSCs was able to induce an over-expression of mature hepatocyte markers, such as ALB, NTCP, and CYP3A4, compared with both 2D hAMSC cultures and the conventional differentiation medium (DM). These data were confirmed by the over-production of ALB protein and over-activity of CYP3A4 observed in organoids grown in 3D hAMSC-CM. Liver repair dysfunction plays a role in the development of liver diseases, and effective repair likely requires the normal functioning of liver stem/progenitor cells. Herein, we showed that hAMSC-CM produced mainly by 3D cultures had the potential to increase hepatic stem/progenitor cell differentiation, demonstrating that soluble factors secreted by those cells are potentially responsible for the reaction. This work shows a potential approach to improve liver repair/regeneration also in a transplantation setting.

Designing bioactive materials, with controlled metal ion release, exerting a significant biological action and associated to low toxicity for humans, is nowadays one of the most important challenges for our community. The most looked-for nanoantimicrobials are capable of releasing metal species with defined kinetic profiles, either by slowing down or inhibiting bacterial growth and pathogenic microorganism diffusion. In this study, laser ablation synthesis in solution (LASiS) has been used to produce bioactive Ag-based nanocolloids, in isopropyl alcohol, which can be used as water-insoluble nano-reservoirs in composite materials like poly(3-hydroxybutyrate-co-3-hydroxyvalerate). Infrared spectroscopy was used to evaluate the chemical state of pristine polymer and final composite material, thus providing useful information about synthesis processes, as well as storage and processing conditions. Transmission electron microscopy was exploited to study the morphology of nano-colloids, along with UV-Vis for bulk chemical characterization, highlighting the presence of spheroidal particles with average diameter around 12 nm. Electro-thermal atomic absorption spectroscopy was used to investigate metal ion release from Ag-modified products, showing a maximum release around 60 ppb, which ensures an efficient antimicrobial activity, being much lower than what recommended by health institutions. Analytical spectroscopy results were matched with bioactivity tests carried out on target microorganisms of food spoilage.

Human mesenchymal stromal/stem cells (MSCs), being immunoprivileged and having immunomodulatory ability, represent a promising tool to be applied in the field of regenerative medicine. Based on numerous in vitro evidences, the immunological effects of MSCs on immune cells could depend on different mechanisms as cell-to-cell contact and paracrine signals. Furthermore, recent studies have shown that the immunomodulatory activity of MSCs is initiated by activated immune cells; thus, their interaction represents a potential homeostatic mechanism by which MSCs regulate the immune response. MSCs also release exosomes able to give different effects, in a paracrine manner, by influencing inflammatory processes. In this study, we aimed to establish the potential role of human amnion-derived MSCs (hAMSCs), in immunomodulation. We found that the immunosuppressive properties of hAMSCs are not constitutive, but require “supportive signals” capable of promoting these properties. Indeed, we observed that hAMSCs alone are not able to produce an adequate amount of soluble immunomodulatory factors. Here, we studied, in depth, the strong immunomodulatory licensing signal deriving from the direct interaction between hAMSCs and stimulated peripheral blood mononuclear cells. We found that the immunomodulatory effect of hAMSCs also depends on cell-to-cell contact through the contribution of the PDL-1/PD-1 axis. We then investigated the IFN-γ priming of hAMSCs (γ-hAMSCs), which induce the increase of PDL-1 expression, high production of IDO, and upregulation of different immunomodulatory exosome-derived miRNAs. Our miRNA–target network analysis revealed that nine of the deregulated miRNAs are involved in the regulation of key proteins that control both T cell activation/anergy and monocyte differentiation pathways. Finally, we observed that γ-hAMSCs induce in monocytes both M2-like phenotype and the increase of IL-10 production. The extensive implications of MSCs in modulating different aspects of the immune system make these cells attractive candidates to be employed in therapeutic application in immune-based diseases. For these reasons, we aimed, with this study, to shed light on the potential of hAMSCs, and how they could become a useful tool for treating different inflammatory diseases, including end-stage pathologies or adverse effects in transplanted patients.

Different approaches have been studied in both preclinical and clinical settings to develop cell-based therapies and/or engineered cell-based therapies to better integrate grafts with the host. In these techniques, much attention is addressed to the use of adult stem cells such as mesenchymal stem cells (MSCs), but identifying and obtaining sufficient numbers of therapeutic cells, and the right route of administration, is often a challenge. In this study, we tested the feasibility of encapsulating human amnion-derived MSCs (hAMSCs) in a semipermeable and biocompatible fiber as a new approach for regenerative medicine. Our data showed that hAMSCs aggregated in the device constitutes an effective system for enhancing, or at least for maintaining, the paracrine activity of these cells in order to better promote tissue regeneration in an immune isolated state. In our new experimental approach, the hAMSCs retained their therapeutic potential, as shown by both the production of specific immunomodulatory/angiogenic factors and immunomodulatory and angiogenic ability observed in vitro. Unlike cell infusion methods, the use of encapsulated-cells leads to minimally invasive approaches, avoiding a direct interaction with the host. Therefore, the potentiality of an allograft or xenograft without the need for immunosuppression, and the lack of tumorigenesis is very intriguing.

Age is the pivotal risk factor for different common medical conditions such as cardiovascular diseases, cancer and dementia. Among age-related disorders, cardiovascular and cerebrovascular diseases, represent the leading causes of premature mortality strictly related to vascular ageing, a pathological condition characterized by endothelial dysfunction, atherosclerosis, hypertension, heart disease and stroke. These features negatively impact on the brain, owing to altered cerebral blood flow, neurovascular coupling and impaired endothelial permeability leading to cerebrovascular diseases (CVDs) as Vascular Dementia (VD) and Parkinsonism (VP). It is an increasing opinion that neurodegenerative disorders and cerebrovascular diseases are associated from a pathogenetic point of view, and in this review, we discuss how cerebrovascular dysfunctions, due to epigenetic alterations, are linked with neuronal degeneration/dysfunction that lead to cognitive impairment. The relation between neurodegenerative and cerebrovascular diseases are reviewed with a focus on role of ncRNAs in age-related vascular diseases impairing the endothelium in the blood-brain barrier with consequent dysfunction of cerebral blood flow. In this review we dissert about different regulatory mechanisms of gene expression implemented by ncRNAs in the pathogenesis of age-related neurovascular impairment, aiming to highlight the potential use of ncRNAs as biomarkers for diagnostic/prognostic purposes as well as novel therapeutic targets.