University of Bologna

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    Fluorogenic hyaluronan-based probe: characterization and use in advanced microscopy

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    Among all, the application of nanomaterials in biomedical research and most recently in the environmental one has opened the fields of nanomedicine and nanoremediation. Sensing methods based on fluorescence optical probe are generally requested for their selectivity, sensitivity. However, most imaging methods in literature rely on a fluorescent covalent labelling of the system. Therefore, the main aim of this project was to synthetise a biocompatible fluorogenic hyaluronan probe (HA) polymer functionalised with a rhomadine B (RB) moieties and study its behaviour as an optical probe with different materials with microscopy techniques. A derivatization of HA with RB (HA-RB) was successfully obtained providing a photophysical characterization showing a particular fluorescence mechanism of the probe. Firstly, we tested the interaction with different lab-grade micro and nanoplastics in water. Thanks to the peculiar photophysical behaviour of the probe nanoplastics can be detected with confocal microscopy and more interestingly their nature can be discriminated based on the fluorescence lifetime decay with FLIM microscopy. After, the interaction of a model plant derived metabolic enzyme GAPC1 undergoing oxidative-triggered aggregation was explored with the HA-RB. We highlighted the probe interaction with the protein even at early stage of the kinetic. Moreover, nanoparticle tracking analysis (NTA) experiment demonstrates that the probe is in fact able to interact with the small pre-aggregates in the early stage of the aggregation kinetic. Ultimately, we focused on the possibility to apply the probe in a super resolution microscopy technique, PALM, exploiting its aspecific interaction to characterize the surface topography of PTFE polydisperse microplastics. Optimal conditions were reached at high concentration of the probe (70 nM) where 0.5-5 nM is always advisable for this technique. Thanks to the polymeric nature and fluorescence mechanism of the probe, this technique was able to reveal features of PTFE surface under the diffraction limit (< 250 nm)

    EBV Type II latency relies on PARP1 activity and is essential for gastric epithelial cell transformation in EBV-associated Gastric Cancer (EBVaGC)

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    Epstein-Barr virus (EBV) establishes a lifelong asymptomatic infection by replicating its chromatinized genome, called episome, together with the host genome. EBV exhibits different latency-associated transcriptional repertoires that mirror its three-dimensional structures of the genome. CTCF, Cohesin and PARP1 are involved in maintaining viral latency and establishing episome architecture. Epstein-Barr virus-associated gastric cancer (EBVaGC) represents almost 10% of all gastric cancers globally. EBVaGC exhibit an intermediate viral transcription profile known as "Latency II", expressing specific viral genes and non-coding RNAs. In this study, we investigated the impact of PARP1 inhibition on CTCF/Cohesin binding in Type II latency. We observed a destabilization of the binding of both factors, leading to a disrupted three-dimensional architecture of the episomes and consequently, an altered viral gene expression. Despite sharing the same CTCF binding profile, Type I, II, and III latencies display different 3D episomal structures that correlate with variations in viral gene expression. Additionally, our analysis of H3K27ac-enriched chromatin interactions revealed differences between Type II latency episomes and a link to cellular transformation through docking of the EBV episomes at specific sites of the Human genome, thus promoting oncogene expression. Overall, this work provides insights into the role of PARP1 in maintaining active latency and novel mechanisms of EBV-induced cellular transformation

    An architecture for network acceleration as a service in the cloud continuum

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    The pervasive availability of connected devices in any industrial and societal sector is pushing for an evolution of the well-established cloud computing model. The emerging paradigm of the cloud continuum embraces this decentralization trend and envisions virtualized computing resources physically located between traditional datacenters and data sources. By totally or partially executing closer to the network edge, applications can have quicker reactions to events, thus enabling advanced forms of automation and intelligence. However, these applications also induce new data-intensive workloads with low-latency constraints that require the adoption of specialized resources, such as high-performance communication options (e.g., RDMA, DPDK, XDP, etc.). Unfortunately, cloud providers still struggle to integrate these options into their infrastructures. That risks undermining the principle of generality that underlies the cloud computing scale economy by forcing developers to tailor their code to low-level APIs, non-standard programming models, and static execution environments. This thesis proposes a novel system architecture to empower cloud platforms across the whole cloud continuum with Network Acceleration as a Service (NAaaS). To provide commodity yet efficient access to acceleration, this architecture defines a layer of agnostic high-performance I/O APIs, exposed to applications and clearly separated from the heterogeneous protocols, interfaces, and hardware devices that implement it. A novel system component embodies this decoupling by offering a set of agnostic OS features to applications: memory management for zero-copy transfers, asynchronous I/O processing, and efficient packet scheduling. This thesis also explores the design space of the possible implementations of this architecture by proposing two reference middleware systems and by adopting them to support interactive use cases in the cloud continuum: a serverless platform and an Industry 4.0 scenario. A detailed discussion and a thorough performance evaluation demonstrate that the proposed architecture is suitable to enable the easy-to-use, flexible integration of modern network acceleration into next-generation cloud platforms

    Transatlantic regulatory divergence in pharmaceuticals: exploring economic and cultural explanations

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    The analysis delves into the regulatory divergence between the EU and the US in the pharmaceutical sector. Through law and economics, political science, and cultural theory perspectives, the study explores how collective preferences within each jurisdiction shape regulations. By identifying cultural orientations such as individualism in the US and hierarchy in the EU, the research suggests that underlying cultural factors play a significant role in regulatory decisions. Despite calls for regulatory harmonization, the recognition of legitimate differences in preferences due to cultural diversity emphasizes that uniformity may not always be the best solution, showcasing the importance of understanding cultural influences in shaping regulatory landscapes

    Black hole scattering from Worldline Quantum Field Theory and the Classical Double Copy of Spinning particles

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    This PhD thesis focuses on studying the classical scattering of massive/massless particles toward black holes, and investigating double copy relations between classical observables in gauge theories and gravity. This is done in the Post-Minkowskian approximation i.e. a perturbative expansion of observables controlled by the gravitational coupling constant κ = 32πGN, with GN being the Newtonian coupling constant. The investigation is performed by using the Worldline Quantum Field Theory (WQFT), displaying a worldline path integral describing the scattering objects and a QFT path integral in the Born approximation, describing the intermediate bosons exchanged in the scattering event by the massive/massless particles. We introduce the WQFT, by deriving a relation between the Kosower- Maybee-O’Connell (KMOC) limit of amplitudes and worldline path integrals, then, we use that to study the classical Compton amplitude and higher point amplitudes. We also present a nice application of our formulation to the case of Hard Thermal Loops (HTL), by explicitly evaluating hard thermal currents in gauge theory and gravity. Next we move to the investigation of the classical double copy (CDC), which is a powerful tool to generate integrands for classical observables related to the binary inspiralling problem in General Relativity. In order to use a Bern-Carrasco-Johansson (BCJ) like prescription, straight at the classical level, one has to identify a double copy (DC) kernel, encoding the locality structure of the classical amplitude. Such kernel is evaluated by using a theory where scalar particles interacts through bi-adjoint scalars. We show here how to push forward the classical double copy so to account for spinning particles, in the framework of the WQFT. Here the quantization procedure on the worldline allows us to fully reconstruct the quantum theory on the gravitational side. Next we investigate how to describe the scattering of massless particles off black holes in the WQFT

    Bridging the gap:exploring the cognitive impact of InterpretBank on Chinese interpreting trainees

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    This exploratory research project developed a cognitive situated approach to studying aspects of simultaneous interpreting with quantitative, confirmatory methods. To do so, it explored how to determine the potential benefits of using a computer-assisted interpreting tool, InterpretBank, among 22 Chinese interpreting trainees with Chinese L1 and English L2. The informants were mostly 2nd-year female students with an average age of 24.7 enrolled in Chinese MA interpreting programs. The study adopted a pretest and posttest design with three cycles. The independent variable was using Excel or InterpretBank. After Cycle I (pre-test), the sample split into control (Excel) and experimental (InterpretBank) groups. Tool choice was compulsory in Cycle II but not Cycle III. The source materials for each cycle were pairs of matching transcripts from popular science podcasts. Informants compiled glossaries out of one transcript, while the other one was edited for simultaneous interpreting, with 39 terms as potential problem triggers. Quantitative profiling results showed that InterpretBank informants spent less time on glossary compilation, generated more terms faster than Excel informants, but their glossaries were less diverse (personal) and longer. The booth tasks yielded no significant differences in fluency indicators except for more bumps (200-600ms silent time gaps) for InterpretBank in Cycle II. InterpretBank informants had more correct renditions in Cycles II and III but there was no statistically significant difference among accuracy indicators per cycle. Holistic quality assessments by PhD raters showed InterpretBank consistently outperforming Excel, suggesting a positive InterpretBank impact on SI quality. However, some InterpretBank implementations raised cognitive ergonomic concerns for Chinese, potentially undermining its utility. Overall, results were mixed regarding InterpretBank benefits for Chinese trainees, but the project was successful in developing cognitive situated interpreting study methods, constructs and indicators

    RNA interference technology as a potential control method for fruit and horticultural crops pathogens Botrytis cinerea and Plasmopara viticola

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    Chapter 1, a general introduction on Botrytis cinerea and its threat to crop production is presented. What Botrytis looks like, its life cycle, why it is a threat to agricultural production, its worldwide pest status, and its current state of management is further elaborated on. Chapter 2, a general introduction on Plasmopara viticola, its threat to grape production and management strategies presented. Chapter 3, titled " RNA Interference Strategies for Future Management of Plant Pathogenic Fungi: Prospects and Challenges ", presents the rapid improvement and extensive implementation of RNA interference (RNAi) technology for the management of fungal pathogens. In this chapter, we describe the application of exogenous RNAi involved in plant pathogenic fungi and discuss dsRNA production, formulation, and RNAi delivery methods. Chapter 4, titled " Exogenous dsRNAs against chitin synthase and glucan synthase genes suppress the growth of the pathogenic fungus Botrytis cinerea " addresses two important questions: Is RNAi technology functional for B. cinerea control ? And which target genes can be exploited for RNAi-based B.cinerea disease control ? Upon target genes selections, an exogenous RNAi protocol was set up and we could effectively deliver a known dose of bacterially produced double stranded RNA (dsRNA) to induce RNAi in B. cinerea. Chapter 5, titled " Double-Stranded RNA Targeting Dicer-Like Genes Compromises the Pathogenicity of Plasmopara viticola on Grapevine “, which deals mainly on RNAi induction against Plasmopara viticola. This chapter addresses two main questions: Is RNAi technology functional in contrasting Plasmopara viticola? And which target genes can be exploited for RNAi-based disease control in Plasmopara viticola?. In the last Chapter (Chapter 6) titled “General discussions and perspectives for future research”, the major research findings from this thesis are discussed together with perspectives for future research

    Il conte di Carmagnola from the last autograph draft to the printing: a new critical edition and a model for a digital edition

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    A seguito degli studi condotti sul manoscritto inedito della copia per la censura della tragedia manzoniana Il Conte di Carmagnola, conservato presso la Houghton Library dell’Università di Harvard (MS Ital 72), il presente lavoro propone una nuova edizione critica delle ultime fasi elaborative dell’opera, dall’ultima redazione autografa (manoscritti Manz.V.S.XI.4 e Manz.B.X.2) alla prima edizione a stampa (1820), muovendosi negli ambiti, ancora scarsamente dialoganti ma complementari, della filologia d’autore e dell’editoria critica digitale. L’edizione, grazie al rinvenimento documentale – già segnalato nel 1990 (Severino 1990), ma ignorato dalla critica fino alla recente menzione di Giulia Raboni (Raboni 2017) – corregge la precedente ricostruzione filologica, sanando alcune erronee interpretazioni causate dalla lacuna testimoniale e rendendo possibile una più precisa ricostruzione delle fasi redazionali dell’opera. La connessa indagine critica degli interventi correttori ascrivibili alle ultime revisioni precedenti la pubblicazione permette inoltre di proporre spunti esegetici più esattamente rispondenti alla progressiva elaborazione del progetto manzoniano, consentendo anche di riassegnare all’autore la quasi totalità delle correzioni intercorse tra autografi e stampa, prima di impossibile o dubbia attribuzione. Lo studio del testo e dei manoscritti, condotto mediante un approccio interdisciplinare volto a misurare le possibilità di integrazione reciproca di filologia tradizionale e digitale, è infine punto di partenza per la realizzazione di un modello di annotazione XML/TEI. Tale schema si offre sia come caso di studio concreto per la rappresentazione del processo genetico della tragedia, anche attraverso la proposta di mockup per la visualizzazione, sia come occasione di riflessione generale sulle possibilità applicative del vocabolario TEI a obiettivi specifici della filologia d’autore, attualmente quasi del tutto esclusa dalle edizioni critiche digitali realizzate.Based on the study of the unpublished manuscript of the censorship copy of Manzoni’s tragedy Il conte di Carmagnola, housed in the Houghton Library of Harvard University (MS Ital 72), this work provides a new critical edition of the last redactions of the tragedy, from the last autograph draft (manuscripts Manz. V.S.XI.4 and Manz.B.X.2) to the first printed edition (1820). The edition – thanks to the analysis of the document, discovered in 1990 (Severino 1990), but ignored by the critics until the recent mention by Giulia Raboni (Raboni 2017) – intends to correct the previous philological reconstruction, by rectifying some misinterpretations due to the witness gap and by presenting a more accurate reconstruction of the editorial phases of the text. Linked to this, a critical investigation of the corrections dating back to the last revisions before the publishing lead me to exegetical observations which respond more likely to the progressive elaboration of the manzonian project and helped me reassigning the author almost the totality of the corrections between autographs and printing which were before of impossible or questionable attribution. The study of the text and the manuscripts – carried out following a multidisciplinary approach which integrates both traditional and digital philology, between which there is a scarce and yet increasingly necessary dialogue – is the starting point for the creation of a XML/TEI encoding model. This scheme offers itself for different purposes. The first is being a concrete case of study for the representation of the genetic process of the tragedy, also through mockup for the visualization. The second is that of being an opportunity for general reflection on application possibilities of TEI vocabulary on specific targets of authorial philology, which is currently underrepresented among digital critical editions

    Investigation of organic semiconductor/water interfaces for optobioelectronic devices

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    The thesis investigates the potential of photoactive organic semiconductors as a new class of materials for developing bioelectronic devices that can convert light into biological signals. The materials can be either small molecules or polymers. When these materials interact with aqueous biological fluids, they give rise to various electrochemical phenomena, including photofaradaic or photocapacitive processes, depending on whether photogenerated charges participate in redox processes or accumulate at an interface. The thesis starts by studying the behavior of the H2Pc/PTCDI molecular p/n thin-film heterojunction in contact with aqueous electrolyte. An equivalent circuit model is developed, explaining the measurements and predicting behavior in wireless mode. A systematic study on p-type polymeric thin-films is presented, comparing rr-P3HT with two low bandgap conjugated polymers: PBDB-T and PTB7. The results demonstrate that PTB7 has superior photocurrent performance due to more effective electron-transfer onto acceptor states in solution. Furthermore, the thesis addresses the issue of photovoltage generation for wireless photoelectrodes. An analytical model based on photoactivated charge-transfer across the organic-semiconductor/water interface is developed, explaining the large photovoltages observed for polymeric p-type semiconductor electrodes in water. Then, flash-precipitated nanoparticles made of the same three photoactive polymers are investigated, assessing the influence of fabrication parameters on the stability, structure, and energetics of the nanoparticles. Photocathodic current generation and consequent positive charge accumulation is also investigated. Additionally, newly developed porous P3HT thin-films are tested, showing that porosity increases both the photocurrent and the semiconductor/water interfacial capacity. Finally, the thesis demonstrates the biocompatibility of the materials in in-vitro experiments and shows safe levels of photoinduced intracellular ROS production with p-type polymeric thin-films and nanoparticles. The findings highlight the potential of photoactive organic semiconductors in the development of optobioelectronic devices, demonstrating their ability to convert light into biological signals and interface with biological fluids

    Diffusing alpha particles radiation treatment optimization in clinical settings

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    Alpha-particle emitters, notably used in 224Ra-DaRT, have emerged as effective in overcoming radiation resistance and providing targeted cancer therapy. These emitters cause DNA double-strand breaks, visualizable in human lymphocytes. The 224Ra DaRT technique, using a decay chain from seeds, extends alpha particle range, achieving complete tumor destruction while sparing healthy tissue. This thesis examines a biokinetic model, validated with patient data, and a feasibility study on skin squamous cell carcinomas are discussed. The study reports 75% tumor complete response rate and 48% patients experiencing acute grade 2 toxicity, resolving within a month. An observed abscopal effect (AE), where tumor regression occurs at non-irradiated sites, is examined, highlighting DaRT's potential in triggering anti-tumor immune responses. This effect, coupled with DaRT's high-linear energy transfer (LET), suggests its superiority over low-LET radiation in certain clinical scenarios. Improvements to DaRT, including the use of an external radio-opaque template for treatment planning, are explored. This advancement aids in determining source numbers for optimal tumor coverage, enhancing DaRT’s safety. The thesis outlines a typical DaRT procedure, from tumor measurements to source assessment and administration, emphasizing the importance of precise seed positioning. Furthermore, the thesis discusses DaRT's potential in treating prostate cancer, a prevalent global health issue, by offering an alternative to traditional salvage therapies. DaRT seeds, delivering alpha particle-based interstitial radiation, require precision in seed insertion due to their limited tissue range. In conclusion, the thesis advocates for DaRT's role in treating solid tumors, emphasizing its improved radiobiological potency and potential benefits over beta and gamma source-based therapies. Ongoing studies are assessing DaRT's feasibility in treating various solid tumors, including pancreatic, breast, prostate, and vulvar malignancies, suggesting a promising future in cancer treatment


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