Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 29

Search results for: Giuseppe Empoli

29 The Influence of Emotional Intelligence Skills on Innovative Start-Ups Coaching: A Neuro-Management Approach

Authors: Alina Parincu, Giuseppe Empoli, Alexandru Capatina

Abstract:

The purpose of this paper is to identify the most influential predictors of emotional intelligence skills, in the case of 20 business innovation coaches, on the co-creation of knowledge through coaching services delivered to innovative start-ups from Europe, funded through Horizon 2020 – SME Instrument. We considered the emotional intelligence skills (self-awareness, self-regulation, motivation, empathy and social skills) as antecedent conditions of the outcome: the quality of coaching services, perceived by the entrepreneurs who received funding within SME instrument, using fuzzy-sets qualitative comparative analysis (fsQCA) approach. The findings reveal that emotional intelligence skills, trained with neuro-management techniques, were associated with increased goal-focused business coaching skills.

Keywords: neuro-management, innovative start-ups, business coaching, fsQCA

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28 Monitoring Systemic Risk in the Hedge Fund Sector

Authors: Frank Hespeler, Giuseppe Loiacono

Abstract:

We propose measures for systemic risk generated through intra-sectorial interdependencies in the hedge fund sector. These measures are based on variations in the average cross-effects of funds showing significant interdependency between their individual returns and the moments of the sector’s return distribution. The proposed measures display a high ability to identify periods of financial distress, are robust to modifications in the underlying econometric model and are consistent with intuitive interpretation of the results.

Keywords: hedge funds, systemic risk, vector autoregressive model, risk monitoring

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27 Aligning Organizational Culture and Compensation Strategies

Authors: Giuseppe Maria Russo, Patrícia Amélia Tomei, Antônio Linhares, André Moreira Santos

Abstract:

Alignment between management strategies, policies and practices with organizational cultures holds great potential to meet the challenges of retaining professionals and maintaining their commitment. In this article, authors consider that when it is aligned with company strategy, compensation acts as an incentive for developing common visions within the organizational culture. This article verified the correlation between types of culture and compensation’s strategic components and provided inputs for the definition of strategies aligned with cultural typologies. We conclude that the impact of compensation variables varies according to the type of organizational culture. This result reinforces the theory that different cultures define different organizational strategies. Thus, compensation strategies may explain types of organizational culture.

Keywords: compensation, Handy’s cultural typology, organizational culture, rewards

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26 Synthesis and Characterization of Model Amines for Corrosion Applications

Authors: John Vergara, Giuseppe Palmese

Abstract:

Fundamental studies aimed at elucidating the key contributions to corrosion performance are needed to make progress toward effective and environmentally compliant corrosion control. Epoxy/amine systems are typically employed as barrier coatings for corrosion control. However, the hardening agents used for coating applications can be very complex, making fundamental studies of water and oxygen permeability challenging to carry out. Creating model building blocks for epoxy/amine coatings is the first step in carrying out these studies. We will demonstrate the synthesis and characterization of model amine building blocks from saturated fatty acids and simple amines such as diethylenetriamine (DETA) and Bis(3-aminopropyl)amine. The structure-property relationship of thermosets made from these model amines and Diglycidyl ether of bisphenol A (DGBEA) will be discussed.

Keywords: building block, amine, synthesis, characterization

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25 The Effects of Key Factors in Traffic-Oriented Road Alignment Adjustment for Low Emissions Profile: A Case Study in Norway

Authors: Gaylord K. Booto, Marinelli Giuseppe, Helge Brattebø, Rolf A. Bohne

Abstract:

Emissions reduction has emerged among the principal targets in the process of planning and designing road alignments today. Intelligent road design methods that can result in optimized alignment constitute concrete and innovative responses towards better alternatives and more sustainable road infrastructures. As the largest amount of emissions of road infrastructures occur in the operation stage, it becomes very important to consider traffic weight and distribution in alignment design process. This study analyzes the effects of four traffic factors (i.e. operating speed, vehicle category, technology and fuel type) on adjusting the vertical alignment of a given road, using optimization techniques. Further, factors’ effects are assessed qualitatively and quantitatively, and the emission profiles of resulting alignment alternatives are compared.

Keywords: alignment adjustment, emissions reduction, optimization, traffic-oriented

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24 A Parallel Approach for 3D-Variational Data Assimilation on GPUs in Ocean Circulation Models

Authors: Rossella Arcucci, Luisa D'Amore, Simone Celestino, Giuseppe Scotti, Giuliano Laccetti

Abstract:

This work is the first dowel in a rather wide research activity in collaboration with Euro Mediterranean Center for Climate Changes, aimed at introducing scalable approaches in Ocean Circulation Models. We discuss designing and implementation of a parallel algorithm for solving the Variational Data Assimilation (DA) problem on Graphics Processing Units (GPUs). The algorithm is based on the fully scalable 3DVar DA model, previously proposed by the authors, which uses a Domain Decomposition approach (we refer to this model as the DD-DA model). We proceed with an incremental porting process consisting of 3 distinct stages: requirements and source code analysis, incremental development of CUDA kernels, testing and optimization. Experiments confirm the theoretic performance analysis based on the so-called scale up factor demonstrating that the DD-DA model can be suitably mapped on GPU architectures.

Keywords: data assimilation, GPU architectures, ocean models, parallel algorithm

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23 Predicting Machine-Down of Woodworking Industrial Machines

Authors: Matteo Calabrese, Martin Cimmino, Dimos Kapetis, Martina Manfrin, Donato Concilio, Giuseppe Toscano, Giovanni Ciandrini, Giancarlo Paccapeli, Gianluca Giarratana, Marco Siciliano, Andrea Forlani, Alberto Carrotta

Abstract:

In this paper we describe a machine learning methodology for Predictive Maintenance (PdM) applied on woodworking industrial machines. PdM is a prominent strategy consisting of all the operational techniques and actions required to ensure machine availability and to prevent a machine-down failure. One of the challenges with PdM approach is to design and develop of an embedded smart system to enable the health status of the machine. The proposed approach allows screening simultaneously multiple connected machines, thus providing real-time monitoring that can be adopted with maintenance management. This is achieved by applying temporal feature engineering techniques and training an ensemble of classification algorithms to predict Remaining Useful Lifetime of woodworking machines. The effectiveness of the methodology is demonstrated by testing an independent sample of additional woodworking machines without presenting machine down event.

Keywords: predictive maintenance, machine learning, connected machines, artificial intelligence

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22 Machine Learning Methods for Flood Hazard Mapping

Authors: Stefano Zappacosta, Cristiano Bove, Maria Carmela Marinelli, Paola di Lauro, Katarina Spasenovic, Lorenzo Ostano, Giuseppe Aiello, Marco Pietrosanto

Abstract:

This paper proposes a novel neural network approach for assessing flood hazard mapping. The core of the model is a machine learning component fed by frequency ratios, namely statistical correlations between flood event occurrences and a selected number of topographic properties. The proposed hybrid model can be used to classify four different increasing levels of hazard. The classification capability was compared with the flood hazard mapping River Basin Plans (PAI) designed by the Italian Institute for Environmental Research and Defence, ISPRA (Istituto Superiore per la Protezione e la Ricerca Ambientale). The study area of Piemonte, an Italian region, has been considered without loss of generality. The frequency ratios may be used as a standalone block to model the flood hazard mapping. Nevertheless, the mixture with a neural network improves the classification power of several percentage points, and may be proposed as a basic tool to model the flood hazard map in a wider scope.

Keywords: flood modeling, hazard map, neural networks, hydrogeological risk, flood risk assessment

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21 Optical Flow Based System for Cross Traffic Alert

Authors: Giuseppe Spampinato, Salvatore Curti, Ivana Guarneri, Arcangelo Bruna

Abstract:

This document describes an advanced system and methodology for Cross Traffic Alert (CTA), able to detect vehicles that move into the vehicle driving path from the left or right side. The camera is supposed to be not only on a vehicle still, e.g. at a traffic light or at an intersection, but also moving slowly, e.g. in a car park. In all of the aforementioned conditions, a driver’s short loss of concentration or distraction can easily lead to a serious accident. A valid support to avoid these kinds of car crashes is represented by the proposed system. It is an extension of our previous work, related to a clustering system, which only works on fixed cameras. Just a vanish point calculation and simple optical flow filtering, to eliminate motion vectors due to the car relative movement, is performed to let the system achieve high performances with different scenarios, cameras and resolutions. The proposed system just uses as input the optical flow, which is hardware implemented in the proposed platform and since the elaboration of the whole system is really speed and power consumption, it is inserted directly in the camera framework, allowing to execute all the processing in real-time.

Keywords: clustering, cross traffic alert, optical flow, real time, vanishing point

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20 Collaborative Writing on Line with Apps During the Time of Pandemic: A Systematic Literature Review

Authors: Giuseppe Liverano

Abstract:

Today’s school iscalledupon to take the lead role in supporting students towards the formation of conscious identity and a sense of responsible citizenship, through the development of key competencies for lifelong learning A rolethatrequiresit to be ready for change and to respond to the ever new needs of students, by adopting new pedagogical and didactic models and new didactic devices. Information and Communication Technologies, in this sense, reveal themselves to be usefulresourcesthatpermit to focus attention on the learning of eachindividualstudentunderstoodas a dynamic and relational process of constructing shared and participatedmeanings. The use of collaborative writing apps represents a democratic and shared knowledge way of constructionthroughICTs. It promotes the learning of reading-writing, literacy, and the development of transversal competencies in an inclusive perspective peer-to-peer comparison and reflectionthatstimulates the transfer of thought into speech and writing, the transformation of knowledge through a trialogicalapproach to learning generates enthusiasm and strengthensmotivationItrepresents a “different” way of expressing the training needs which come from several disciplinary fields of subjects with different cultures. The contribution aims to reflect on the formative value of collaborative writing through apps and analyse some proposals on line at school during the time of pandemic in order to highlight their critical aspects and pedagogical perspectives.

Keywords: collaborative writing, formative value, online, apps, pandemic

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19 Smart Side View Mirror Camera for Real Time System

Authors: Nunziata Ivana Guarneri, Arcangelo Bruna, Giuseppe Spampinato, Antonio Buemi

Abstract:

In the last decade, automotive companies have invested a lot in terms of innovation about many aspects regarding the automatic driver assistance systems. One innovation regards the usage of a smart camera placed on the car’s side mirror for monitoring the back and lateral road situation. A common road scenario is the overtaking of the preceding car and, in this case, a brief distraction or a loss of concentration can lead the driver to undertake this action, even if there is an already overtaking vehicle, leading to serious accidents. A valid support for a secure drive can be a smart camera system, which is able to automatically analyze the road scenario and consequentially to warn the driver when another vehicle is overtaking. This paper describes a method for monitoring the side view of a vehicle by using camera optical flow motion vectors. The proposed solution detects the presence of incoming vehicles, assesses their distance from the host car, and warns the driver through different levels of alert according to the estimated distance. Due to the low complexity and computational cost, the proposed system ensures real time performances.

Keywords: camera calibration, ego-motion, Kalman filters, object tracking, real time systems

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18 Enhancing Disaster Response Capabilities in Asia-Pacific: An Explorative Study Applied to Decision Support Tools for Logistics Network Design

Authors: Giuseppe Timperio, Robert de Souza

Abstract:

Logistics operations in the context of disaster response are characterized by a high degree of complexity due to the combined effect of a large number of stakeholders involved, time pressure, uncertainties at various levels, massive deployment of goods and personnel, and gigantic financial flow to be managed. It also involves several autonomous parties such as government agencies, militaries, NGOs, UN agencies, private sector to name few, to have a highly collaborative approach especially in the critical phase of the immediate response. This is particularly true in the context of L3 emergencies that are the most severe, large-scale humanitarian crises. Decision-making processes in disaster management are thus extremely difficult due to the presence of multiple decision-makers involved, and the complexity of the tasks being tackled. Hence, in this paper, we look at applying ICT based solutions to enable a speedy and effective decision making in the golden window of humanitarian operations. A high-level view of ICT based solutions in the context of logistics operations for humanitarian response in Southeast Asia is presented, and their viability in a real-life case about logistics network design is explored.

Keywords: decision support, disaster preparedness, humanitarian logistics, network design

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17 Axial, Bending Interaction Diagrams of Reinforced Concrete Columns Exposed to Chloride Attack

Authors: Rita Greco, Giuseppe Carlo Marano

Abstract:

Chloride induced reinforcement corrosion is widely accepted to be the most frequent mechanism causing premature degradation of reinforced concrete members, whose economic and social consequences are growing up continuously. Prevention of these phenomena has a great importance in structural design, and modern Codes and Standard impose prescriptions concerning design details and concrete mix proportion for structures exposed to different external aggressive conditions, grouped in environmental classes. This paper focuses on reinforced concrete columns load carrying capacity degradation over time due to chloride induced steel pitting corrosion. The structural element is considered to be exposed to marine environment and the effects of corrosion are described by the time degradation of the axial-bending interaction diagram. Because chlorides ingress and consequent pitting corrosion propagation are both time-dependent mechanisms, the study adopts a time-variant predictive approach to evaluate the residual strength of corroded reinforced concrete columns at different lifetimes. Corrosion initiation and propagation process is modelled by taking into account all the parameters, such as external environmental conditions, concrete mix proportion, concrete cover and so on, which influence the time evolution of the corrosion phenomenon and its effects on the residual strength of RC columns.

Keywords: pitting corrosion, strength deterioration, diffusion coefficient, surface chloride concentration, concrete structures, marine environment

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16 A Network Optimization Study of Logistics for Enhancing Emergency Preparedness in Asia-Pacific

Authors: Giuseppe Timperio, Robert De Souza

Abstract:

The combination of factors such as temperamental climate change, rampant urbanization of risk exposed areas, political and social instabilities, is posing an alarming base for the further growth of number and magnitude of humanitarian crises worldwide. Given the unique features of humanitarian supply chain such as unpredictability of demand in space, time, and geography, spike in the number of requests for relief items in the first days after the calamity, uncertain state of logistics infrastructures, large volumes of unsolicited low-priority items, a proactive approach towards design of disaster response operations is needed to achieve high agility in mobilization of emergency supplies in the immediate aftermath of the event. This paper is an attempt in that direction, and it provides decision makers with crucial strategic insights for a more effective network design for disaster response. Decision sciences and ICT are integrated to analyse the robustness and resilience of a prepositioned network of emergency strategic stockpiles for a real-life case about Indonesia, one of the most vulnerable countries in Asia-Pacific, with the model being built upon a rich set of quantitative data. At this aim, a network optimization approach was implemented, with several what-if scenarios being accurately developed and tested. Findings of this study are able to support decision makers facing challenges related with disaster relief chains resilience, particularly about optimal configuration of supply chain facilities and optimal flows across the nodes, while considering the network structure from an end-to-end in-country distribution perspective.

Keywords: disaster preparedness, humanitarian logistics, network optimization, resilience

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15 Molecularly Imprinted Nanoparticles (MIP NPs) as Non-Animal Antibodies Substitutes for Detection of Viruses

Authors: Alessandro Poma, Kal Karim, Sergey Piletsky, Giuseppe Battaglia

Abstract:

The recent increasing emergency threat to public health of infectious influenza diseases has prompted interest in the detection of avian influenza virus (AIV) H5N1 in humans as well as animals. A variety of technologies for diagnosing AIV infection have been developed. However, various disadvantages (costs, lengthy analyses, and need for high-containment facilities) make these methods less than ideal in their practical application. Molecularly Imprinted Polymeric Nanoparticles (MIP NPs) are suitable to overcome these limitations by having high affinity, selectivity, versatility, scalability and cost-effectiveness with the versatility of post-modification (labeling – fluorescent, magnetic, optical) opening the way to the potential introduction of improved diagnostic tests capable of providing rapid differential diagnosis. Here we present our first results in the production and testing of MIP NPs for the detection of AIV H5N1. Recent developments in the solid-phase synthesis of MIP NPs mean that for the first time a reliable supply of ‘soluble’ synthetic antibodies can be made available for testing as potential biological or diagnostic active molecules. The MIP NPs have the potential to detect viruses that are widely circulating in farm animals and indeed humans. Early and accurate identification of the infectious agent will expedite appropriate control measures. Thus, diagnosis at an early stage of infection of a herd or flock or individual maximizes the efficiency with which containment, prevention and possibly treatment strategies can be implemented. More importantly, substantiating the practicability’s of these novel reagents should lead to an initial reduction and eventually to a potential total replacement of animals, both large and small, to raise such specific serological materials.

Keywords: influenza virus, molecular imprinting, nanoparticles, polymers

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14 Preliminary Short-Term Results of a Population of Patients Treated with Mitraclip Therapy: One Center Experience

Authors: Rossana Taravella, Gilberto M. Cellura, Giuseppe Cirrincione, Salvatore Asciutto, Marco Caruso, Massimo Benedetto, Renato Ciofalo, Giuliana Pace, Salvatore Novo

Abstract:

Objectives: This retrospective analysis sought to evaluate 1-month outcomes and therapy effectiveness of a population of patients treated with MitraClip therapy. We describe in this article the preliminary results of primary effectiveness endpoint. Background: Percutaneous Mitral Repair is being developed to treat severe mitral regurgitation (MR), with increasing real-world cases of functional MR (FMR). In the EVEREST (Endovascular Valve Edge-to-Edge Repair Study)II trial, the percutaneous device showed superior safety but less reduction in MR at 1year. 4-year outcomes from EVEREST II trial showed no difference in the prevalence of moderate-severe and severe MR or mortality at 4years between surgical mitral repair and percutaneous approach. Methods: We analysed retrospectively collected data from one center experience in Italy enrolled from January 2011 to December 2016. The study included 62 patients [mean age 74±11years, 43 men (69%)] with MR of at least grade3+. Most of the patients had functional MR, were in New York Heart Association (NYHA) functional class III or IV, with a large portion (78%) of mild-to-moderate Tricuspid Regurgitation (TR). One or more clips were implanted in 67 procedures (62 patients). Results and Conclusions: Severity of MR was reduced in all successfully treated patients,54(90%) were discharged with MR≤2+ (primary effectiveness endpoint). Clinical 1-month follow-up data showed an improvement in NYHA functional class (42 patients (70%) in NYHA class I-II). 60 of 62 (97 %) successfully treated patients were free from death and mitral valve surgery at 1-month follow-up. MitraClip therapy reduces functional MR with acute MR reduction to <2+ in the great majority of patients, with a large freedom from death, surgery or recurrent MR in a great portion of patients.

Keywords: MitraClip, mitral regurgitation, heart valves, catheter-based therapy

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13 The Contact between a Rigid Substrate and a Thick Elastic Layer

Authors: Nicola Menga, Giuseppe Carbone

Abstract:

Although contact mechanics has been widely focused on the study of contacts between half-space, it has been recently pointed out that in presence of finite thickness elastic layers the results of the contact problem show significant difference in terms of the main contact quantities (e.g. contact area, penetration, mean pressure, etc.). Actually, there exist a wide range of industrial application demanding for this kind of studies, such as seals leakage prediction or pressure-sensitive coatings for electrical applications. In this work, we focus on the contact between a rigid profile and an elastic layer of thickness h confined under two different configurations: rigid constrain and applied uniform pressure. The elastic problem at hand has been formalized following Green’s function method and then numerically solved by means of a matrix inversion. We study different contact conditions, both considering and neglecting adhesive interactions at the interface. This leads to different solution techniques: Adhesive contacts equilibrium solution is found, in term of contact area for given penetration, making stationary the total free energy of the system; whereas, adhesiveless contacts are addressed defining an equilibrium criterion, again on the contact area, relying on the fracture mechanics stress intensity factor KI. In particular, we make the KI vanish at the edges of the contact area, as peculiar for adhesiveless elastic contacts. The results are obtained in terms of contact area, penetration, and mean pressure for both adhesive and adhesiveless contact conditions. As expected, in the case of a uniform applied pressure the slab turns out much more compliant than the rigidly constrained one. Indeed, we have observed that the peak value of the contact pressure, for both the adhesive and adhesiveless condition, is much higher for the rigidly constrained configuration than in the case of applied uniform pressure. Furthermore, we observed that, for little contact area, both systems behave the same and the pull-off occurs at approximately the same contact area and mean contact pressure. This is an expected result since in this condition the ratio between the layers thickness and the contact area is very high and both layer configurations recover the half-space behavior where the pull-off occurrence is mainly controlled by the adhesive interactions, which are kept constant among the cases.

Keywords: contact mechanics, adhesion, friction, thick layer

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12 Sound Source Localisation and Augmented Reality for On-Site Inspection of Prefabricated Building Components

Authors: Jacques Cuenca, Claudio Colangeli, Agnieszka Mroz, Karl Janssens, Gunther Riexinger, Antonio D'Antuono, Giuseppe Pandarese, Milena Martarelli, Gian Marco Revel, Carlos Barcena Martin

Abstract:

This study presents an on-site acoustic inspection methodology for quality and performance evaluation of building components. The work focuses on global and detailed sound source localisation, by successively performing acoustic beamforming and sound intensity measurements. A portable experimental setup is developed, consisting of an omnidirectional broadband acoustic source and a microphone array and sound intensity probe. Three main acoustic indicators are of interest, namely the sound pressure distribution on the surface of components such as walls, windows and junctions, the three-dimensional sound intensity field in the vicinity of junctions, and the sound transmission loss of partitions. The measurement data is post-processed and converted into a three-dimensional numerical model of the acoustic indicators with the help of the simultaneously acquired geolocation information. The three-dimensional acoustic indicators are then integrated into an augmented reality platform superimposing them onto a real-time visualisation of the spatial environment. The methodology thus enables a measurement-supported inspection process of buildings and the correction of errors during construction and refurbishment. Two experimental validation cases are shown. The first consists of a laboratory measurement on a full-scale mockup of a room, featuring a prefabricated panel. The latter is installed with controlled defects such as lack of insulation and joint sealing material. It is demonstrated that the combined acoustic and augmented reality tool is capable of identifying acoustic leakages from the building defects and assist in correcting them. The second validation case is performed on a prefabricated room at a near-completion stage in the factory. With the help of the measurements and visualisation tools, the homogeneity of the partition installation is evaluated and leakages from junctions and doors are identified. Furthermore, the integration of acoustic indicators together with thermal and geometrical indicators via the augmented reality platform is shown.

Keywords: acoustic inspection, prefabricated building components, augmented reality, sound source localization

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11 Influence of the Popularity of Opera during Risorgimento on Foreign Presence in Italy

Authors: Andrew Wee

Abstract:

As a result of the Italian Independence Wars starting in 1848, Italy began to change through unification. People gradually moved away from some of their traditional practices and values, such as the long-held belief that women were inferior to men, as part of the Risorgimento. Italians began to take interest in opera as a form of emotional release. As opera became more popular and prominent in their culture, it aided in the dissemination of ideas, especially stimulating the spread of imperialism, in the late 19th century, as Italy began extending its presence to other countries. In order to collect the information needed to analyze Italy’s foreign presence, it was necessary to consult texts concerning the culture of the Risorgimento. These texts included primary sources from operatic composers and contemporary recorded accounts. Letters from Giuseppe Verdi, a leader in opera during the Risorgimento, have been scrutinized for indications of popular attitudes of the time. The cultural context of the Risorgimento is essential to understanding the Italian motives and attitudes towards the outside world. On the more political side, research has also entailed the study of historical data of general laws, policies, and their purposes concerning geopolitical boundaries and foreign affairs, such as Edward Said’s thesis on Orientalism. By establishing these two characteristics of Italy, the paper will thoroughly illustrate Italy’s presence in foreign affairs. Texts have been searched with the intent of using information that reveals Italian attitudes toward exotic countries to determine whether their demeanor was positive or condescending. Motives behind sources have been interpreted in context in order to form a complete picture of the Italian sentiment towards foreigners. Additionally, research pertaining to Italian nationalism and imperialism such as song and literature has been used. The primary form of research has been the division of sources that are culturally based and those that are political in nature. Opera had always been developing since its creation in the 17th century, and in the 19th century, the bel canto movement revolutionized opera and its role in Italian society. This paper uses evidence that popular sentiment was influenced by opera to support the belief that the evolution of opera was as a result of the nationalist sentiment, and in turn fueled the cultural movement known as the Risorgimento. In this way, opera proceeded to affect Italian culture by spreading the idea of imperialism.

Keywords: opera, Italian unification, music history, imperialism

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10 Mirna Expression Profile is Different in Human Amniotic Mesenchymal Stem Cells Isolated from Obese Respect to Normal Weight Women

Authors: Carmela Nardelli, Laura Iaffaldano, Valentina Capobianco, Antonietta Tafuto, Maddalena Ferrigno, Angela Capone, Giuseppe Maria Maruotti, Maddalena Raia, Rosa Di Noto, Luigi Del Vecchio, Pasquale Martinelli, Lucio Pastore, Lucia Sacchetti

Abstract:

Maternal obesity and nutrient excess in utero increase the risk of future metabolic diseases in the adult life. The mechanisms underlying this process are probably based on genetic, epigenetic alterations and changes in foetal nutrient supply. In mammals, the placenta is the main interface between foetus and mother, it regulates intrauterine development, modulates adaptive responses to sub optimal in uterus conditions and it is also an important source of human amniotic mesenchymal stem cells (hA-MSCs). We previously highlighted a specific microRNA (miRNA) profiling in amnion from obese (Ob) pregnant women, here we compared the miRNA expression profile of hA-MSCs isolated from (Ob) and control (Co) women, aimed to search for any alterations in metabolic pathways that could predispose the new-born to the obese phenotype. Methods: We isolated, at delivery, hA-MSCs from amnion of 16 Ob- and 7 Co-women with pre-pregnancy body mass index (mean/SEM) 40.3/1.8 and 22.4/1.0 kg/m2, respectively. hA-MSCs were phenotyped by flow cytometry. Globally, 384 miRNAs were evaluated by the TaqMan Array Human MicroRNA Panel v 1.0 (Applied Biosystems). By the TargetScan program we selected the target genes of the miRNAs differently expressed in Ob- vs Co-hA-MSCs; further, by KEGG database, we selected the statistical significant biological pathways. Results: The immunophenotype characterization confirmed the mesenchymal origin of the isolated hA-MSCs. A large percentage of the tested miRNAs, about 61.4% (232/378), was expressed in hA-MSCs, whereas 38.6% (146/378) was not. Most of the expressed miRNAs (89.2%, 207/232) did not differ between Ob- and Co-hA-MSCs and were not further investigated. Conversely, 4.8% of miRNAs (11/232) was higher and 6.0% (14/232) was lower in Ob- vs Co-hA-MSCs. Interestingly, 7/232 miRNAs were obesity-specific, being expressed only in hA-MSCs isolated from obese women. Bioinformatics showed that these miRNAs significantly regulated (P<0.001) genes belonging to several metabolic pathways, i.e. MAPK signalling, actin cytoskeleton, focal adhesion, axon guidance, insulin signaling, etc. Conclusions: Our preliminary data highlight an altered miRNA profile in Ob- vs Co-hA-MSCs and suggest that an epigenetic miRNA-based mechanism of gene regulation could affect pathways involved in placental growth and function, thereby potentially increasing the newborn’s risk of metabolic diseases in the adult life.

Keywords: hA-MSCs, obesity, miRNA, biosystem

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9 Acrylate-Based Photopolymer Resin Combined with Acrylated Epoxidized Soybean Oil for 3D-Printing

Authors: Raphael Palucci Rosa, Giuseppe Rosace

Abstract:

Stereolithography (SLA) is one of the 3D-printing technologies that has been steadily growing in popularity for both industrial and personal applications due to its versatility, high accuracy, and low cost. Its printing process consists of using a light emitter to solidify photosensitive liquid resins layer-by-layer to produce solid objects. However, the majority of the resins used in SLA are derived from petroleum and characterized by toxicity, stability, and recalcitrance to degradation in natural environments. Aiming to develop an eco-friendly resin, in this work, different combinations of a standard commercial SLA resin (Peopoly UV professional) with a vegetable-based resin were investigated. To reach this goal, different mass concentrations (varying from 10 to 50 wt%) of acrylated epoxidized soybean oil (AESO), a vegetable resin produced from soyabean oil, were mixed with a commercial acrylate-based resin. 1.0 wt% of Diphenyl(2,4,6-trimethylbenzoyl) phosphine oxide (TPO) was used as photo-initiator, and the samples were printed using a Peopoly moai 130. The machine was set to operate at standard configurations when printing commercial resins. After the print was finished, the excess resin was drained off, and the samples were washed in isopropanol and water to remove any non-reacted resin. Finally, the samples were post-cured for 30 min in a UV chamber. FT-IR analysis was used to confirm the UV polymerization of the formulated resin with different AESO/Peopoly ratios. The signals from 1643.7 to 1616, which corresponds to the C=C stretching of the AESO acrylic acids and Peopoly acrylic groups, significantly decreases after the reaction. The signal decrease indicates the consumption of the double bonds during the radical polymerization. Furthermore, the slight change of the C-O-C signal from 1186.1 to 1159.9 decrease of the signals at 809.5 and 983.1, which corresponds to unsaturated double bonds, are both proofs of the successful polymerization. Mechanical analyses showed a decrease of 50.44% on tensile strength when adding 10 wt% of AESO, but it was still in the same range as other commercial resins. The elongation of break increased by 24% with 10 wt% of AESO and swelling analysis showed that samples with a higher concentration of AESO mixed absorbed less water than their counterparts. Furthermore, high-resolution prototypes were printed using both resins, and visual analysis did not show any significant difference between both products. In conclusion, the AESO resin was successful incorporated into a commercial resin without affecting its printability. The bio-based resin showed lower tensile strength than the Peopoly resin due to network loosening, but it was still in the range of other commercial resins. The hybrid resin also showed better flexibility and water resistance than Peopoly resin without affecting its resolution. Finally, the development of new types of SLA resins is essential to provide new sustainable alternatives to the commercial petroleum-based ones.

Keywords: 3D-printing, bio-based, resin, soybean, stereolithography

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8 A Comparative Study of the Tribological Behavior of Bilayer Coatings for Machine Protection

Authors: Cristina Diaz, Lucia Perez-Gandarillas, Gonzalo Garcia-Fuentes, Simone Visigalli, Roberto Canziani, Giuseppe Di Florio, Paolo Gronchi

Abstract:

During their lifetime, industrial machines are often subjected to chemical, mechanical and thermal extreme conditions. In some cases, the loss of efficiency comes from the degradation of the surface as a result of its exposition to abrasive environments that can cause wear. This is a common problem to be solved in industries of diverse nature such as food, paper or concrete industries, among others. For this reason, a good selection of the material is of high importance. In the machine design context, stainless steels such as AISI 304 and 316 are widely used. However, the severity of the external conditions can require additional protection for the steel and sometimes coating solutions are demanded in order to extend the lifespan of these materials. Therefore, the development of effective coatings with high wear resistance is of utmost technological relevance. In this research, bilayer coatings made of Titanium-Tantalum, Titanium-Niobium, Titanium-Hafnium, and Titanium-Zirconium have been developed using magnetron sputtering configuration by PVD (Physical Vapor Deposition) technology. Their tribological behavior has been measured and evaluated under different environmental conditions. Two kinds of steels were used as substrates: AISI 304, AISI 316. For the comparison with these materials, titanium alloy substrate was also employed. Regarding the characterization, wear rate and friction coefficient were evaluated by a tribo-tester, using a pin-on-ball configuration with different lubricants such as tomato sauce, wine, olive oil, wet compost, a mix of sand and concrete with water and NaCl to approximate the results to real extreme conditions. In addition, topographical images of the wear tracks were obtained in order to get more insight of the wear behavior and scanning electron microscope (SEM) images were taken to evaluate the adhesion and quality of the coating. The characterization was completed with the measurement of nanoindentation hardness and elastic modulus. Concerning the results, thicknesses of the samples varied from 100 nm (Ti-Zr layer) to 1.4 µm (Ti-Hf layer) and SEM images confirmed that the addition of the Ti layer improved the adhesion of the coatings. Moreover, results have pointed out that these coatings have increased the wear resistance in comparison with the original substrates under environments of different severity. Furthermore, nanoindentation hardness results showed an improvement of the elastic strain to failure and a high modulus of elasticity (approximately 200 GPa). As a conclusion, Ti-Ta, Ti-Zr, Ti-Nb, and Ti-Hf are very promising and effective coatings in terms of tribological behavior, improving considerably the wear resistance and friction coefficient of typically used machine materials.

Keywords: coating, stainless steel, tribology, wear

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7 University Building: Discussion about the Effect of Numerical Modelling Assumptions for Occupant Behavior

Authors: Fabrizio Ascione, Martina Borrelli, Rosa Francesca De Masi, Silvia Ruggiero, Giuseppe Peter Vanoli

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The refurbishment of public buildings is one of the key factors of energy efficiency policy of European States. Educational buildings account for the largest share of the oldest edifice with interesting potentialities for demonstrating best practice with regards to high performance and low and zero-carbon design and for becoming exemplar cases within the community. In this context, this paper discusses the critical issue of dealing the energy refurbishment of a university building in heating dominated climate of South Italy. More in detail, the importance of using validated models will be examined exhaustively by proposing an analysis on uncertainties due to modelling assumptions mainly referring to the adoption of stochastic schedules for occupant behavior and equipment or lighting usage. Indeed, today, the great part of commercial tools provides to designers a library of possible schedules with which thermal zones can be described. Very often, the users do not pay close attention to diversify thermal zones and to modify or to adapt predefined profiles, and results of designing are affected positively or negatively without any alarm about it. Data such as occupancy schedules, internal loads and the interaction between people and windows or plant systems, represent some of the largest variables during the energy modelling and to understand calibration results. This is mainly due to the adoption of discrete standardized and conventional schedules with important consequences on the prevision of the energy consumptions. The problem is surely difficult to examine and to solve. In this paper, a sensitivity analysis is presented, to understand what is the order of magnitude of error that is committed by varying the deterministic schedules used for occupation, internal load, and lighting system. This could be a typical uncertainty for a case study as the presented one where there is not a regulation system for the HVAC system thus the occupant cannot interact with it. More in detail, starting from adopted schedules, created according to questioner’ s responses and that has allowed a good calibration of energy simulation model, several different scenarios are tested. Two type of analysis are presented: the reference building is compared with these scenarios in term of percentage difference on the projected total electric energy need and natural gas request. Then the different entries of consumption are analyzed and for more interesting cases also the comparison between calibration indexes. Moreover, for the optimal refurbishment solution, the same simulations are done. The variation on the provision of energy saving and global cost reduction is evidenced. This parametric study wants to underline the effect on performance indexes evaluation of the modelling assumptions during the description of thermal zones.

Keywords: energy simulation, modelling calibration, occupant behavior, university building

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6 Energy Refurbishment of University Building in Cold Italian Climate: Energy Audit and Performance Optimization

Authors: Fabrizio Ascione, Martina Borrelli, Rosa Francesca De Masi, Silvia Ruggiero, Giuseppe Peter Vanoli

Abstract:

The Directive 2010/31/EC 'Directive of the European Parliament and of the Council of 19 may 2010 on the energy performance of buildings' moved the targets of the previous version toward more ambitious targets, for instance by establishing that, by 31 December 2020, all new buildings should demand nearly zero-energy. Moreover, the demonstrative role of public buildings is strongly affirmed so that also the target nearly zero-energy buildings is anticipated, in January 2019. On the other hand, given the very low turn-over rate of buildings (in Europe, it ranges between 1-3%/yearly), each policy that does not consider the renovation of the existing building stock cannot be effective in the short and medium periods. According to this proposal, the study provides a novel, holistic approach to design the refurbishment of educational buildings in colder cities of Mediterranean regions enabling stakeholders to understand the uncertainty to use numerical modelling and the real environmental and economic impacts of adopting some energy efficiency technologies. The case study is a university building of Molise region in the centre of Italy. The proposed approach is based on the application of the cost-optimal methodology as it is shown in the Delegate Regulation 244/2012 and Guidelines of the European Commission, for evaluating the cost-optimal level of energy performance with a macroeconomic approach. This means that the refurbishment scenario should correspond to the configuration that leads to lowest global cost during the estimated economic life-cycle, taking into account not only the investment cost but also the operational costs, linked to energy consumption and polluting emissions. The definition of the reference building has been supported by various in-situ surveys, investigations, evaluations of the indoor comfort. Data collection can be divided into five categories: 1) geometrical features; 2) building envelope audit; 3) technical system and equipment characterization; 4) building use and thermal zones definition; 5) energy building data. For each category, the required measures have been indicated with some suggestions for the identifications of spatial distribution and timing of the measurements. With reference to the case study, the collected data, together with a comparison with energy bills, allowed a proper calibration of a numerical model suitable for the hourly energy simulation by means of EnergyPlus. Around 30 measures/packages of energy, efficiency measure has been taken into account both on the envelope than regarding plant systems. Starting from results, two-point will be examined exhaustively: (i) the importance to use validated models to simulate the present performance of building under investigation; (ii) the environmental benefits and the economic implications of a deep energy refurbishment of the educational building in cold climates.

Keywords: energy simulation, modelling calibration, cost-optimal retrofit, university building

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5 The Last National Anthem of the Ottoman Empire: Musical Code, Sociopolitical Control and Historical Realities

Authors: Nuray Ocakli

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19th century was the era of changes and transformations for the Ottoman Empire. The first sultan of this century, Mahmud II (1808-1839), was the architect of Ottoman modernization and fundamental changes. The most radical of these was abolishing the Janissary corps and the traditional Ottoman military band, Mehteran. Mahmud II introduced modernized military corps as well as western style royal and military music. Mahmut II invited the Italian composer Giuseppe Donizetti to establish a modern military band for the new army and to compose the Sultan’s royal anthem. In 1828, Donizetti composed the first western-style Ottoman anthem, Mahmudiyye anthem. During the 19th and early 20th century, four other western style Ottoman anthems (Aziziyye, Mecidiyye, Hamidiyye, and Resadiyye) were composed but the last anthem adopted in the reign of Mehmet VI (r. 1918-1922) was again Mahmudiyye anthem. This paper aims to analyze the Mahmudiyye anthem composed as royal anthem in 1828 but adopted as national anthem in 1918. Research questions of this paper are as follows: What were the characteristics of the Mahmudiyye anthem making it the best choice of the last sultan for the last national anthem? Are there specific reasons of the last sultan to adopt Mahmudiyye anthem or not to adopt any of the other four anthems? The musical characteristics of the anthem are analyzed based on the Cerulo’s empirical research. Cerulo examined the musical structures of 124 western style anthems from 150 countries in the 1580-1976 period. Cerulo’s research categorizes musical codes of the anthems as basic and embellished related with the level of sociopolitical control. Musical analysis of the anthem indicates that the basic musical code of the anthem implies a high level of socio-political control during the reign of both Mahmut II and Mehmet VI. Historical analysis of each sultans’ reign shows that both sultans were autocratic. Mahmut II designed authoritarian government policies to suppress possible reactions against his reforms. On the other hand, authoritarian policies of Mehmet VI are related with the domestic and international political conditions following the World War I. Historical analysis of the research questions show that compared to the other western style Ottoman anthems, Mahmudiyye anthem remained the only neutral anthem symbolizing modernization and westernization of the empire. Other anthems were all the symbols of failed ideologies such as Ottomanism, pan-Islamism, and pan-Turkism. In the early 20th century, there were a few common things remained among the diverse communities of the Ottoman Empire: The land they shared as homeland and the idea of modernization to save the homeland. For this reason, the last sultan Mehmet VI adopted Mahmudiyye anthem as the memory of a unified empire under the rule of a powerful and modernist sultan. The last sultan’s reign lasted just for four years, and the Ottoman Empire disintegrated in 1922, but his adaptation of the Mahmudiyye anthem indicates his unifying policies, his attitudes to save the empire and the caliphate.

Keywords: Mahmudiyye anthem, musical code, national anthem, Ottoman Empire, royal anthem

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4 Corrosion Protective Coatings in Machines Design

Authors: Cristina Diaz, Lucia Perez, Simone Visigalli, Giuseppe Di Florio, Gonzalo Fuentes, Roberto Canziani, Paolo Gronchi

Abstract:

During the last 50 years, the selection of materials is one of the main decisions in machine design for different industrial applications. It is due to numerous physical, chemical, mechanical and technological factors to consider in it. Corrosion effects are related with all of these factors and impact in the life cycle, machine incidences and the costs for the life of the machine. Corrosion affects the deterioration or destruction of metals due to the reaction with the environment, generally wet. In food industry, dewatering industry, concrete industry, paper industry, etc. corrosion is an unsolved problem and it might introduce some alterations of some characteristics in the final product. Nowadays, depending on the selected metal, its surface and its environment of work, corrosion prevention might be a change of metal, use a coating, cathodic protection, use of corrosion inhibitors, etc. In the vast majority of the situations, use of a corrosion resistant material or in its defect, a corrosion protection coating is the solution. Stainless steels are widely used in machine design, because of their strength, easily cleaned capacity, corrosion resistance and appearance. Typical used are AISI 304 and AISI 316. However, their benefits don’t fit every application, and some coatings are required against corrosion such as some paintings, galvanizing, chrome plating, SiO₂, TiO₂ or ZrO₂ coatings, etc. In this work, some coatings based in a bilayer made of Titanium-Tantalum, Titanium-Niobium, Titanium-Hafnium or Titanium-Zirconium, have been developed used magnetron sputtering configuration by PVD (Physical Vapor Deposition) technology, for trying to reduce corrosion effects on AISI 304, AISI 316 and comparing it with Titanium alloy substrates. Ti alloy display exceptional corrosion resistance to chlorides, sour and oxidising acidic media and seawater. In this study, Ti alloy (99%) has been included for comparison with coated AISI 304 and AISI 316 stainless steel. Corrosion tests were conducted by a Gamry Instrument under ASTM G5-94 standard, using different electrolytes such as tomato salsa, wine, olive oil, wet compost, a mix of sand and concrete with water and NaCl for testing corrosion in different industrial environments. In general, in all tested environments, the results showed an improvement of corrosion resistance of all coated AISI 304 and AISI 316 stainless steel substrates when they were compared to uncoated stainless steel substrates. After that, comparing these results with corrosion studies on uncoated Ti alloy substrate, it was observed that in some cases, coated stainless steel substrates, reached similar current density that uncoated Ti alloy. Moreover, Titanium-Zirconium and Titanium-Tantalum coatings showed for all substrates in study including coated Ti alloy substrates, a reduction in current density more than two order in magnitude. As conclusion, Ti-Ta, Ti-Zr, Ti-Nb and Ti-Hf coatings have been developed for improving corrosion resistance of AISI 304 and AISI 316 materials. After corrosion tests in several industry environments, substrates have shown improvements on corrosion resistance. Similar processes have been carried out in Ti alloy (99%) substrates. Coated AISI 304 and AISI 316 stainless steel, might reach similar corrosion protection on the surface than uncoated Ti alloy (99%). Moreover, coated Ti Alloy (99%) might increase its corrosion resistance using these coatings.

Keywords: coatings, corrosion, PVD, stainless steel

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3 Microsimulation of Potential Crashes as a Road Safety Indicator

Authors: Vittorio Astarita, Giuseppe Guido, Vincenzo Pasquale Giofre, Alessandro Vitale

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Traffic microsimulation has been used extensively to evaluate consequences of different traffic planning and control policies in terms of travel time delays, queues, pollutant emissions, and every other common measured performance while at the same time traffic safety has not been considered in common traffic microsimulation packages as a measure of performance for different traffic scenarios. Vehicle conflict techniques that were introduced at intersections in the early traffic researches carried out at the General Motor laboratory in the USA and in the Swedish traffic conflict manual have been applied to vehicles trajectories simulated in microscopic traffic simulators. The concept is that microsimulation can be used as a base for calculating the number of conflicts that will define the safety level of a traffic scenario. This allows engineers to identify unsafe road traffic maneuvers and helps in finding the right countermeasures that can improve safety. Unfortunately, most commonly used indicators do not consider conflicts between single vehicles and roadside obstacles and barriers. A great number of vehicle crashes take place with roadside objects or obstacles. Only some recent proposed indicators have been trying to address this issue. This paper introduces a new procedure based on the simulation of potential crash events for the evaluation of safety levels in microsimulation traffic scenarios, which takes into account also potential crashes with roadside objects and barriers. The procedure can be used to define new conflict indicators. The proposed simulation procedure generates with the random perturbation of vehicle trajectories a set of potential crashes which can be evaluated accurately in terms of DeltaV, the energy of the impact, and/or expected number of injuries or casualties. The procedure can also be applied to real trajectories giving birth to new surrogate safety performance indicators, which can be considered as “simulation-based”. The methodology and a specific safety performance indicator are described and applied to a simulated test traffic scenario. Results indicate that the procedure is able to evaluate safety levels both at the intersection level and in the presence of roadside obstacles. The procedure produces results that are expressed in the same unity of measure for both vehicle to vehicle and vehicle to roadside object conflicts. The total energy for a square meter of all generated crash can be used and is shown on the map, for the test network, after the application of a threshold to evidence the most dangerous points. Without any detailed calibration of the microsimulation model and without any calibration of the parameters of the procedure (standard values have been used), it is possible to identify dangerous points. A preliminary sensitivity analysis has shown that results are not dependent on the different energy thresholds and different parameters of the procedure. This paper introduces a specific new procedure and the implementation in the form of a software package that is able to assess road safety, also considering potential conflicts with roadside objects. Some of the principles that are at the base of this specific model are discussed. The procedure can be applied on common microsimulation packages once vehicle trajectories and the positions of roadside barriers and obstacles are known. The procedure has many calibration parameters and research efforts will have to be devoted to make confrontations with real crash data in order to obtain the best parameters that have the potential of giving an accurate evaluation of the risk of any traffic scenario.

Keywords: road safety, traffic, traffic safety, traffic simulation

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2 Affordable and Environmental Friendly Small Commuter Aircraft Improving European Mobility

Authors: Diego Giuseppe Romano, Gianvito Apuleo, Jiri Duda

Abstract:

Mobility is one of the most important societal needs for amusement, business activities and health. Thus, transport needs are continuously increasing, with the consequent traffic congestion and pollution increase. Aeronautic effort aims at smarter infrastructures use and in introducing greener concepts. A possible solution to address the abovementioned topics is the development of Small Air Transport (SAT) system, able to guarantee operability from today underused airfields in an affordable and green way, helping meanwhile travel time reduction, too. In the framework of Horizon2020, EU (European Union) has funded the Clean Sky 2 SAT TA (Transverse Activity) initiative to address market innovations able to reduce SAT operational cost and environmental impact, ensuring good levels of operational safety. Nowadays, most of the key technologies to improve passenger comfort and to reduce community noise, DOC (Direct Operating Costs) and pilot workload for SAT have reached an intermediate level of maturity TRL (Technology Readiness Level) 3/4. Thus, the key technologies must be developed, validated and integrated on dedicated ground and flying aircraft demonstrators to reach higher TRL levels (5/6). Particularly, SAT TA focuses on the integration at aircraft level of the following technologies [1]: 1)    Low-cost composite wing box and engine nacelle using OoA (Out of Autoclave) technology, LRI (Liquid Resin Infusion) and advance automation process. 2) Innovative high lift devices, allowing aircraft operations from short airfields (< 800 m). 3) Affordable small aircraft manufacturing of metallic fuselage using FSW (Friction Stir Welding) and LMD (Laser Metal Deposition). 4)       Affordable fly-by-wire architecture for small aircraft (CS23 certification rules). 5) More electric systems replacing pneumatic and hydraulic systems (high voltage EPGDS -Electrical Power Generation and Distribution System-, hybrid de-ice system, landing gear and brakes). 6) Advanced avionics for small aircraft, reducing pilot workload. 7) Advanced cabin comfort with new interiors materials and more comfortable seats. 8) New generation of turboprop engine with reduced fuel consumption, emissions, noise and maintenance costs for 19 seats aircraft. (9) Alternative diesel engine for 9 seats commuter aircraft. To address abovementioned market innovations, two different platforms have been designed: Reference and Green aircraft. Reference aircraft is a virtual aircraft designed considering 2014 technologies with an existing engine assuring requested take-off power; Green aircraft is designed integrating the technologies addressed in Clean Sky 2. Preliminary integration of the proposed technologies shows an encouraging reduction of emissions and operational costs of small: about 20% CO2 reduction, about 24% NOx reduction, about 10 db (A) noise reduction at measurement point and about 25% DOC reduction. Detailed description of the performed studies, analyses and validations for each technology as well as the expected benefit at aircraft level are reported in the present paper.

Keywords: affordable, European, green, mobility, technologies development, travel time reduction

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1 In vivo Evaluation of LAB Probiotic Potential with the Zebrafish Animal Model

Authors: Iñaki Iturria, Pasquale Russo, Montserrat Nacher-Vázquez, Giuseppe Spano, Paloma López, Miguel Angel Pardo

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Introduction: It is known that some Lactic Acid Bacteria (LAB) present an interesting probiotic effect. Probiotic bacteria stimulate host resistance to microbial pathogens and thereby aid in immune response, and modulate the host's immune responses to antigens with a potential to down-regulate hypersensitivity reactions. Therefore, probiotic therapy is valuable against intestinal infections and may be beneficial in the treatment of Inflammatory Bowel Disease (IBD). Several in vitro tests are available to evaluate the probiotic potential of a LAB strain. However, an in vivo model is required to understand the interaction between the host immune system and the bacteria. During the last few years, zebrafish (Danio rerio) has gained interest as a promising vertebrate model in this field. This organism has been extensively used to study the interaction between the host and the microbiota, as well as the host immune response under several microbial infections. In this work, we report on the use of the zebrafish model to investigate in vivo the colonizing ability and the immunomodulatory effect of probiotic LAB. Methods: Lactobacillus strains belonging to different LAB species were fluorescently tagged and used to colonize germ-free zebrafish larvae gastrointestinal tract (GIT). Some of the strains had a well-documented probiotic effect (L. acidophilus LA5); while others presented an exopolysaccharide (EPS) producing phenotype, thus allowing evaluating the influence of EPS in the colonization and immunomodulatory effect. Bacteria colonization was monitored for 72 h by direct observation in real time using fluorescent microscopy. CFU count per larva was also evaluated at different times. The immunomodulatory effect was assessed analysing the differential expression of several innate immune system genes (MyD88, NF-κB, Tlr4, Il1β and Il10) by qRT- PCR. The anti-inflammatory effect was evaluated using a chemical enterocolitis zebrafish model. The protective effect against a pathogen was also studied. To that end, a challenge test was developed using a fluorescently tagged pathogen (Vibrio anguillarum-GFP+). The progression of the infection was monitored up to 3 days using a fluorescent stereomicroscope. Mortality rates and CFU counts were also registered. Results and conclusions: Larvae exposed to EPS-producing bacteria showed a higher fluorescence and CFU count than those colonized with no-EPS phenotype LAB. In the same way, qRT-PCR results revealed an immunomodulatory effect on the host after the administration of the strains with probiotic activity. A downregulation of proinflammatory cytoquines as well as other cellular mediators of inflammation was observed. The anti-inflammatory effect was found to be particularly marked following exposure to LA% strain, as well as EPS producing strains. Furthermore, the challenge test revealed a protective effect of probiotic administration. As a matter of fact, larvae fed with probiotics showed a decrease in the mortality rate ranging from 20 to 35%. Discussion: In this work, we developed a promising model, based on the use of gnotobiotic zebrafish coupled with a bacterial fluorescent tagging in order to evaluate the probiotic potential of different LAB strains. We have successfully used this system to monitor in real time the colonization and persistence of exogenous LAB within the gut of zebrafish larvae, to evaluate their immunomodulatory effect and for in vivo competition assays. This approach could bring further insights into the complex microbial-host interactions at intestinal level.

Keywords: gnotobiotic, immune system, lactic acid bacteria, probiotics, zebrafish

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