LA PROCURA EUROPEA NEL CONTESTO DEL PROCESSO DI INTEGRAZIONE IN MATERIA PENALE TRA VINCOLI NAZIONALI E SOVRANAZIONALI

La tesi affronta il tema della realizzazione della Procura europea (European Public Prosecutor\u2019s Office, acronimo EPPO), istituita con il regolamento (UE) 2017/1939 del Consiglio del 12 ottobre 2017, dopo quasi un ventennio di studi e proposte e non senza significative rinunce rispetto alle originarie aspirazioni. Tradizionalmente il tema \ue8 stato affrontato dagli studiosi attraverso l\u2019impiego di categorie concettuali proprie dei settori disciplinari del diritto penale e del diritto processuale penale, con tutti i meriti, ma ovviamente anche i limiti, di un\u2019indagine che comporta l\u2019adozione di una prospettiva di diritto interno. La predilezione per un simile approccio si deve, sostanzialmente, alla genesi del progetto che si \ue8 sviluppato a partire da uno studio comparatistico, ma soprattutto alle sue implicazioni. Comprensibilmente, infatti, la dottrina italiana e straniera si \ue8 interessata principalmente della sua compatibilit\ue0 con gli ordinamenti nazionali, misurando la fattibilit\ue0 e realizzabilit\ue0 del progetto con le eterogeneit\ue0 proprie delle tradizioni giuridiche degli stessi. L\u2019elaborato inquadra, invece, il tema trattato dal punto di osservazione del diritto dell\u2019Unione europea sul presupposto l\u2019Ufficio \ue8 destinato ad inserirsi nel contesto giuridico-istituzionale dell\u2019UE, nel rispetto dei principi che fondano la competenza dell\u2019UE in materia penale, ne regolamentano l\u2019esercizio e, infine, stabiliscono la portata e l\u2019intensit\ue0 dell\u2019azione dell\u2019UE in tale \u201cdelicato\u201d settore. Nella medesima ottica, merita altrettanta considerazione il fatto che EPPO si collocher\ue0 nel panorama degli attori ed organismi di cooperazione e coordinamento gi\ue0 esistenti, aventi specifiche competenze in materia penale ed attivi anche nel campo della protezione degli interessi finanziari dell\u2019UE. In tale prospettiva, il primo capitolo intende costituire la premessa metodologica e la chiave di lettura dell\u2019intero lavoro, inquadrando l\u2019argomento trattato nel contesto delle specificit\ue0 del processo di integrazione europea in materia penale, con particolare attenzione ai suoi principi ispiratori ed ai suoi connaturati limiti. Il secondo capitolo ripercorre gli studi, le proposte, i documenti (istituzionali e non) che hanno dato avvio al dibattito intorno alla figura di un\u2019autorit\ue0 inquirente europea e che hanno condotto all\u2019introduzione di una base giuridica ad hoc per l\u2019istituzione dell\u2019ufficio - inserita dal trattato di Lisbona all\u2019art. 86 TFUE \u2013 sul cui fondamento \ue8 stato intrapreso un lungo e complesso iter legislativo conclusosi, in data 12 ottobre 2017, con l\u2019approvazione del regolamento. Il terzo, quarto e quinto capitolo sono dedicati, rispettivamente, ai profili istituzionali, alla competenza materiale e, da ultimo, agli aspetti operativi relativi al funzionamento dell\u2019Ufficio ed ai rapporti con i \u201cpartners\u201d. Le previsioni del testo di regolamento sono esaminate mettendone in evidenza i profili critici e potenzialmente problematici. Il lavoro indaga anche le specifiche implicazioni connesse al ricorso alla cooperazione rafforzata.The thesis deals with the issue of the creation of the European Public Prosecutor's Office (EPPO), established by Council Regulation (EU) 2017/1939 of 12 October 2017, after almost two decades of studies and proposals. Traditionally, the subject has been studied in the disciplinary fields of criminal law and criminal procedure, with all the merits but also all the limits of a research that involves the adoption of a perspective of domestic law. The predilection for such an approach was essentially due to the genesis of the project that developed from a comparative study, but above all to its implications. Reasonably, in fact, the Italian and foreign doctrine was mainly concerned with its compatibility with national laws and national legal traditions. Unlike these studies, the paper examines the subject from the perspective of EU law. In fact, EPPO set-up regulation must respect the principles that base the competence of the EU in criminal matters, regulate the exercise and finally establish the scope and the intensity of EU action in this "sensitive" sector. Furthermore, the Office will place itself in the panorama of the already existing cooperation and coordination actors and bodies, having specific competences in criminal matters and active also in the field of the protection of the financial interests of the EU. From this perspective, the first chapter represents the methodological premise and the key to understanding the entire work. In this chapter the issue is framed in the context of the specificities of the process of European integration in criminal matters, with particular attention to its inspiring principles and its inherent limits. The second chapter examines the studies, proposals, documents (both institutional and non-institutional) that started the debate around the figure of a European investigating authority and which led to the introduction of an ad hoc legal basis for the establishment of the Office, inserted by the Treaty of Lisbon in the art. 86 TFEU. The chapter ends with the long and complex legislative process that led to the approval of the regulation even if with significant renunciations compared to the original aspirations. First of all, the use of enhanced cooperation. The third, fourth and fifth chapters are dedicated, respectively, to institutional profiles, material competence and, lastly, to the operational aspects related to the functioning of the Office and to relations with "partners". In this part, the provisions of the regulation text are examined by highlighting the critical and potentially problematic profiles. The work also investigates the specific implications related to the use of enhanced cooperation

Simulated hypergravity induces changes in human tendon-derived cells: from cell morphology to gene expression

Gravity influences physical and biological processes, having an impact on development, as well as homeostasis of living systems. The musculoskeletal system is comprised of several mechano- responsive tissues and altered gravitational forces are known to influence distinct properties, including bone mineral density and skeletal muscle mass. This is particularly relevant in a near- weightlessness (microgravity) environment, which is found during spaceflight and, not less importantly, during bed resting. Over the years, several studies have been conducted under simulated conditions of altered gravity owing to the advances on ground-based facilities, such as bioreactors for microgravity / hypo-gravity (1g) studies. Interestingly, microgravity-induced alterations are comparable to tissue degeneration caused by disuse and ageing. In turn, exposing musculoskeletal tissues to hypergravity may constitute a way of simulating (over)loading or, eventually, to be used as a measure to rescue cell phenotype after exposure to near-weightlessness conditions. Different studies have focused on bone, cartilage and skeletal muscle, but effects on tendons and ligaments have been underappreciated. Therefore, we evaluated the influence of increasing g-levels (5g, 10g, 15g and 20g) and different hypergravity exposure periods (4 and 16 h) on the behaviour of human tendon- derived cells (hTDCs). For this purpose, hTDCs were exposed to simulated hypergravity conditions using the Large Diameter Centrifuge (LDC) from the European Space Research and Technology Centre (ESTEC, ESA, The Netherlands). Human TDCs cultured under standard conditions (1g, normogravity, Earth gravity force) were used as controls. The effects of hypergravity on the viability of hTDCs, as well as on the expression of tendon related markers at the gene level were evaluated. Simulated hypergravity resulted in a reduced cell content after 16 h independently of g-level, as determined by DNA quantification. Additionally, the different g-levels studied led to changes in cell and cytoskeleton morphology. Strikingly, a 16-hour period of exposure resulted in alterations of gene expression profiles. Overall, gene expression of tendon-related markers, including collagen types I (col1a1) and III (col3a1), scleraxis (scx), tenomodulin (tnmd), decorin (dcn) and tenascin (tnc), seemed to be increased upon hypergravity stimulation and in comparison to cells cultured under control conditions. Altogether, these results highlight that altered gravity, particularly simulated hypergravity, has an influence on the phenotype of tendon cells, opening new avenues for research focused on using altered gravity as a model for overloading-induced tendon tissue injury or as measure to rescue the phenotype of degenerated tendon cells. Acknowledgements The authors would like to thank ESA Education Office for Spin Your Thesis! 2016 programme. R.C-A acknowledges the PhD grant SFRH/BD/96593/2013 from FCT â Fundação para a Ciência e a Tecnologia. SFRH/BD/96593/2013 from FCT –Fundação para a Ciência e a Tecnologiainfo:eu-repo/semantics/publishedVersio

Rapid production of human liver scaffolds for functional tissue engineering by high shear stress oscillation-decellularization

The development of human liver scaffolds retaining their 3-dimensional structure and extra-cellular matrix (ECM) composition is essential for the advancement of liver tissue engineering. We report the design and validation of a new methodology for the rapid and accurate production of human acellular liver tissue cubes (ALTCs) using normal liver tissue unsuitable for transplantation. The application of high shear stress is a key methodological determinant accelerating the process of tissue decellularization while maintaining ECM protein composition, 3D-architecture and physico-chemical properties of the native tissue. ALTCs were engineered with human parenchymal and non-parenchymal liver cell lines (HepG2 and LX2 cells, respectively), human umbilical vein endothelial cells (HUVEC), as well as primary human hepatocytes and hepatic stellate cells. Both parenchymal and non-parenchymal liver cells grown in ALTCs exhibited markedly different gene expression when compared to standard 2D cell cultures. Remarkably, HUVEC cells naturally migrated in the ECM scaffold and spontaneously repopulated the lining of decellularized vessels. The metabolic function and protein synthesis of engineered liver scaffolds with human primary hepatocytes reseeded under dynamic conditions were maintained. These results provide a solid basis for the establishment of effective protocols aimed at recreating human liver tissue in vitro

The Future of Personalized Medicine in Space: From Observations to Countermeasures

The aim of personalized medicine is to detach from a “one-size fits all approach” and improve patient health by individualization to achieve the best outcomes in disease prevention, diagnosis and treatment. Technological advances in sequencing, improved knowledge of omics, integration with bioinformatics and new in vitro testing formats, have enabled personalized medicine to become a reality. Individual variation in response to environmental factors can affect susceptibility to disease and response to treatments. Space travel exposes humans to environmental stressors that lead to physiological adaptations, from altered cell behavior to abnormal tissue responses, including immune system impairment. In the context of human space flight research, human health studies have shown a significant inter-individual variability in response to space analogue conditions. A substantial degree of variability has been noticed in response to medications (from both an efficacy and toxicity perspective) as well as in susceptibility to damage from radiation exposure and in physiological changes such as loss of bone mineral density and muscle mass in response to deconditioning. At present, personalized medicine for astronauts is limited. With the advent of longer duration missions beyond low Earth orbit, it is imperative that space agencies adopt a personalized strategy for each astronaut, starting from pre-emptive personalized pre-clinical approaches through to individualized countermeasures to minimize harmful physiological changes and find targeted treatment for disease. Advances in space medicine can also be translated to terrestrial applications, and vice versa. This review places the astronaut at the center of personalized medicine, will appraise existing evidence and future preclinical tools as well as clinical, ethical and legal considerations for future space travel

Site Specific Modification of Adeno-Associated Virus Enables Both Fluorescent Imaging of Viral Particles and Characterization of the Capsid Interactome

Adeno-associated viruses (AAVs) are attractive gene therapy vectors due to their low toxicity, high stability, and rare integration into the host genome. Expressing ligands on the viral capsid can re-target AAVs to new cell types, but limited sites have been identified on the capsid that tolerate a peptide insertion. Here, we incorporated a site-specific tetracysteine sequence into the AAV serotype 9 (AAV9) capsid, to permit labelling of viral particles with either a fluorescent dye or biotin. We demonstrate that fluorescently labelled particles are detectable in vitro, and explore the utility of the method in vivo in mice with time-lapse imaging. We exploit the biotinylated viral particles to generate two distinct AAV interactomes, and identify several functional classes of proteins that are highly represented: actin/cytoskeletal protein binding, RNA binding, RNA splicing/processing, chromatin modifying, intracellular trafficking and RNA transport proteins. To examine the biological relevance of the capsid interactome, we modulated the expression of two proteins from the interactomes prior to AAV transduction. Blocking integrin αVβ6 receptor function reduced AAV9 transduction, while reducing histone deacetylase 4 (HDAC4) expression enhanced AAV transduction. Our method demonstrates a strategy for inserting motifs into the AAV capsid without compromising viral titer or infectivity

Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein

A method for non-invasive visualization of genetically labelled cells in animal disease models with micron-level resolution would greatly facilitate development of cell-based therapies. Imaging of fluorescent proteins (FPs) using red excitation light in the “optical window” above 600 nm is one potential method for visualizing implanted cells. However, previous efforts to engineer FPs with peak excitation beyond 600 nm have resulted in undesirable reductions in brightness. Here we report three new red-excitable monomeric FPs obtained by structure-guided mutagenesis of mNeptune, previously the brightest monomeric FP when excited beyond 600 nm. Two of these, mNeptune2 and mNeptune2.5, demonstrate improved maturation and brighter fluorescence, while the third, mCardinal, has a red-shifted excitation spectrum without reduction in brightness. We show that mCardinal can be used to non-invasively and longitudinally visualize the differentiation of myoblasts and stem cells into myocytes in living mice with high anatomical detail