1,720,979 research outputs found
Development of novel strategies for measuring extracellular ATP dynamics
No full textLa segnalazione purinergica rappresenta un sofisticato meccanismo di risposta cellulare mediato dalle purine, tra le quali ATP, ADP, UDP e adenosina. Questo sistema di segnalazione svolge un ruolo decisivo in numerosi processi fisiologici, consentendo alle cellule di comunicare e rispondere in modo efficiente agli stimoli esterni. In particolare, l'ATP extracellulare riveste una funzione centrale nella segnalazione purinergica e nei processi infiammatori da essa mediata, regolando sia fenomeni fisiologici che patologici. In quanto molecola chiave della risposta infiammatoria e precursore dell'adenosina, un potente agente immunosoppressivo, l'ATP extracellulare è coinvolto in numerosi meccanismi nell’ambito del sistema nervoso centrale, della maturazione delle cellule immunitarie, del microambiente tumorale e della sepsi. Da tale premessa nasce la necessità di disporre di strumenti affidabili per misurare l'ATP extracellulare in vitro e soprattutto in vivo, per comprendere meglio il suo ruolo fisiologico e patologico.
Questa tesi presenta due sistemi innovativi per la misurazione dell'ATP extracellulare: la sonda raziometrica pmeLUC/nilla e un costrutto nanobody-luciferasi. La pmeLUC/nilla rappresenta un'evoluzione della sonda pmeLUC, ormai largamente utilizzata, e consiste in un vettore bicistronico contenente due luciferasi, la pmeLUC e la Renilla luciferasi, separate da un elemento IRES. Questo sistema permette la misurazione normalizzata dell'ATP extracellulare, risultando indipendente dall'efficienza di trasfezione e dai livelli di espressione e garantendo una maggiore accuratezza in diversi contesti sperimentali.
Il secondo sistema descritto, un costrutto nanobody-luciferasi, è stato progettato in modo da legare il recettore P2X7 e ha prodotto risultati promettenti per la rilevazione dell’ATP extracellulare in vitro. Il costrutto denominato 7E2opt-dim-lucYY5 ha mostrato elevata specificità per le cellule HEK-293 che esprimono il recettore P2X7 murino, permettendo di correlare accuratamente la luminescenza emessa alla concentrazione di ATP. Questo sistema presenta un notevole vantaggio rispetto ad altre sonde per la misurazione dell'ATP, in quanto non richiede trasfezione ed è pronto all'uso. Inoltre, si programma in futuro di testare questa sonda in vivo.
Complessivamente, questi due sistemi rappresentano un importante progresso nella misurazione dell'ATP extracellulare, con potenziali applicazioni nel campo dell'oncologia, dell'immunologia e in genere nella ricerca biomedica. Questi strumenti potrebbero contribuire all'ambizioso progetto di mappare le fluttuazioni dell'ATP extracellulare nell'organismo, offrendo nuove prospettive per comprendere più a fondo il suo ruolo nei processi fisiologici e patologici.Purinergic signaling represents a sophisticated cellular response mechanism mediated by purine compounds, particularly ATP, ADP, UDP, and adenosine. This signaling pathway plays a crucial role in various physiological processes, allowing cells to communicate and respond effectively to changes in their environment. Extracellular ATP plays a central role in purinergic signaling and inflammation, influencing several physiological and pathological processes. As a key inflammatory signaling molecule and precursor of the immunosuppressive adenosine, extracellular ATP is involved in critical mechanisms in the central nervous system, tumor microenvironment, immune cell maturation, and the onset of sepsis, among others. This highlights the need for reliable tools to measure extracellular ATP in vitro and, more importantly, in vivo, to better understand its involvement in health and disease.
This thesis presents two innovative systems for measuring extracellular ATP: the pmeLUC/nilla ratiometric probe and a nanobody-luciferase construct. pmeLUC/nilla is an improvement of the widely used pmeLUC probe and consist of a bicistronic vector containing two luciferases, the pmeLUC and the Renilla luciferase separated by an IRES element. The pmeLUC/nilla allows the normalized measurement of extracellular ATP, thus it is independent from transfection efficiency and expression levels. Thanks to this feature this system enables a more accurate measurement of extracellular ATP in several research contexts.
The second system is a nanobody-luciferase construct that targets the P2X7 receptor and has shown promising results for in vitro ATP detection. The construct named 7E2opt-dim-lucYY5 demonstrated high specificity for HEK-293 cells expressing murine P2X7 and provided reliable luminescence readings, dependent on ATP concentration. This system offers substantial advantages over the other probes for ATP detection because it does not require transfection being a ready-to-use tool. It will be further characterized for an in vivo use.
Together, these systems provide substantial advancements in extracellular ATP measurement, with potential applications in cancer biology, immunology, and broader fields of biomedical research. Ultimately, these tools may contribute to the ambitious goal of mapping extracellular ATP fluctuations throughout the body, facilitating deeper insights into its role in health and disease
LCA (Life Cycle Assessment) of roses and cyclamens in greenhouse cultivation
No full textThe flower market of the Terlizzi agro-industrial district (Bari, Italy) represents the 20% of the Italian production, which is the second in Europe after the Netherlands. The main products are cut flowers and plants in vase, in particular roses and cyclamens. In this framework the European Project "Ecoflower Terlizzi" was carried out to support the implementation of a sustainable environmental policy in the Terlizzi district. The project was aimed at defining the criteria of environmental quality of the flowers produced in this area and includes both the definition of an EPD (Environmental Product Declaration) programme and of a local eco-label (type I). To reach this objective environmental analyses and LCA (Life Cycle Assessment) studies were carried out on a sample of seven enterprises representative of the Terlizzi's production systems. They were analysed on the basis of on-site data collection including materials and components for structures and equipments, energy and water consumption, fertilizers, pesticides and other chemicals used in the cultivation phase. Three farms produce roses with soilless cultivation systems, two produce roses in soil and two produce cyclamens' pots. In this paper the main issues concerning the application of the LCA methodology to the production of roses and cyclamens and the results obtained will be presented. Thermal energy consumption gives the main contribution to the environmental impacts of the roses' life cycle. For the cyclamens' pots the main contribution comes from the seedlings production. For both productions, electricity consumption, structures and equipments give a significant contribution to the environmental impacts
Analysis of efforts needed to modify LCI databases to be compliant with two data quality requirements - Final Technical Report
No full textThe final technical report documents the course of the project, including organisational and administrative issues. This report comprehends an executive summary describing main results, conclusions, recommendations and the monthly progress emails (project diary)
Analysis of efforts needed to modify LCI databases to be compliant with two data quality requirements – Methodology Report
No full textThis document is the deliverable of Task 1 Define the methodology to be used to analyse a sample of datasets and to extrapolate the findings to the database level” of the Service Contract 388957 between JRC IES and ENEA on the subject “Analysis of efforts needed to modify LCI databases to be compliant with two data quality requirements”.
This report is focussed on the demonstration of the methodology adopted to analyse the compliance of the datasets and database, defined and detailed in the Technical Offer to the Tender and further specified in the Inception Report, using two datasets: one unit process (from Ecoinvent) and one aggregated (from Gabi).
After a description of the goal and scope of the methodology test, chapter 3 focuses on compliance criteria. Its purpose is to point out and discuss possible inconsistencies in the quality criteria, raised during the discussion with the databases’ developers. Moreover, the feasibility of evaluating the scores for all the 6 criteria is discussed. Then, chapter 4 goes into the detail of the methodology test, by describing first how the datasets for the analysis have been selected, then discussing the structure and business model of the two databases (very important for both the analysis of the compliance and the estimation of the effort for solving identified non compliances) and finally reporting the results of the test. Chapter 5 closes the report, providing some recommendations for the next phases of the analysis
The Mediterranean Eco-Industrial Development Model
No full textIndustrial areas are a fundamental element of territorial development, since they can promote growth, jobs and entrepreneurship. Nevertheless the excessive exploitation of raw materials, the use of non-renewable energy resources, the emission of pollutants and waste production have up to now represented a weakness in the present production model. In this context Mediterranean Eco-Industrial Development (MEID) project, funded by the European program MED, aims to define a joint model to plan, build and manage sustainable Industrial Areas in the Mediterranean region, improving sustainable development and SMEs competitiveness. The model intends to enhance capacities and develop decision tools for Competent Authorities and Industrial Area Managers to integrate environmental friendly solutions into the Regional and Interregional Industrial Development Strategies. An incremental approach has been adopted to ensure the model applicability to new industrial areas, as well as already operating non structured and structured areas. Fundamental parts of the management model are high level infrastructures and innovative services to support SMEs to create networks, exploit the eco-innovation opportunities and face the challenges of the Green economy
Necessary steps and efforts to make existing LCI databases meet the ILCD Data Network entry level requirements and PEF data requirements Rev 1
No full textThis document is the deliverable of Task 2 “Apply the methodology to analyse a sample of datasets and to extrapolate the findings to the database level to the selected datasets / databases” of the Service Contract 388957 between JRC IES and ENEA on the subject “Analysis of efforts needed to modify LCI databases to be compliant with two data quality requirements”.
The report is focused on the results obtained by the application of the methodology adopted to analyse the compliance of datasets and databases, defined and detailed in the methodological report (Report ENEA UTVALAMB P9VF 013), and applied to three commercial databases: GaBi, Ecoinvent and EIME, as reported in ENEA UTVALAMB P9VF 016, ENEA UTVALAMB P9VF 017 and ENEA UTVALAMB P9VF 018, respectively.
After a description of the aim of the document and the goal and scope of the analysis, chapter 3 focuses on the method used. Its purpose is to describe how the results of the compliance analysis on the three databases have been generalized. Then, chapter 4 goes into the detail of the obtained results and, starting from the estimated effort needed to make each database compliant with the two quality requirements (ILCD entry level and PEF), proposes a road map for their implementation, considering the differences and commonalities among the databases. An estimation of the required efforts is also provided when available. Finally, Chapter 5 closes the report, providing some general conclusions and recommendations.
In Appendix, proposals of changes for PEF data quality indicators and rating are suggested.
This revision 1 of the report includes the comments received by JRC and the databases’ providers
Analysis of efforts needed to modify LCI databases to be comliant with two data quality requirements – Interim Technical Report
No full textThe interim technical report documents the course of the project, including organisational and administrative issues. It is composed of the monthly progress emails. This report comprehends a short summary and an adjusted project work-flow chart of the completed and of the foreseen tasks and deliverables of the project as well as timing
Life Cycle Assessment of waste management systems in Italian industrial areas: Case study of 1st Macrolotto of Prato
No full textLife Cycle Assessment (LCA) is not widely used as a decision supporting tool in Italy, despite recent European Commission policies foster its adoption to achieve an energy and resource efficient economy. In this paper, a LCA of waste management system of 1st Macrolotto industrial area is presented. The aims of the study were to identify the environmental critical points of the system and to evaluate opportunities and problems in applying this methodology at industrial area level. After a description of the waste management system of 1st Macrolotto industrial area, the main assumptions of the study and some elements of the system modelling are presented. Results confirmed that door-to-door separated collection and packaging recycling are successful strategies for reducing the use of natural resources. The cooperation of the waste management company, which was part of the area management structure, was strategic for the completion of the analysis. Country specific databases and models should be developed for a widespread application of the LCA methodology to waste management in Italian industrial areas
Towards Sustainable Industrial Areas: building a Mediterranean model
No full textAn EU research project (Mediterranean Eco-Industrial Development-MEID) which involves different actors from six Mediterranean regions is currently in progress. Its objective is to provide a decision support tool (model) to plan, build and govern more competitive Sustainable Industrial Areas (SIA) in this region. Indeed, well planned and equipped Industrial Areas (IAs) stimulate the relocation of industries and help to relieve congestion and pollution in metropolitan areas. In order to correspond to a coherent solution a SWOT analysis of several target IAs is performed. Following its results, it is possible to define the most relevant aspects aiming at sustainability of IAs in the Mediterranean area. In this paper the general context and the first results of the MEID project are presented
Analysis of efforts needed to modify LCI databases to be compliant with two data quality requirements – Inception Report
No full textThe European Commission, Joint Research Centers, Institute of Environmental Science (JRC IES) awarded ENEA with the Service Contract 388957 on the subject “Analysis of efforts needed to modify LCI databases to be compliant with two data quality requirements”. Start date of the project is December 12th 2012, defined as T0 in the contract and its annexes.
On February 21st 2013, in ISPRA at the JRC IES premises, the kickoff meeting of the project was held, aimed at presenting, discussing and agreeing upon administrative, technical and magerial aspects of the project. In particular, the methodology proposed in the offer (annex III of the contract) was further detailed by ENEA. This document, Inception Report, further details the concepts and procedure proposed in the offer taking into account the outcome of the kickoff meeting
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