The concept of decentralised international cooperation promotes the involvement of sub-national actors, both local communities and local authorities, in the governance of transboundary natural resources and spaces. This concept describes a global legal phenomenon that can be observed in different regions of the world wherever decentralised cooperative mechanisms are at work. These mechanisms are context-specific and tailor cooperation to geographical peculiarities and local needs. They have a transboundary but localised spatial dimension that is ecologically functional, complements inter-state cooperation, and enables the participation of sub-national actors across borders. This thesis explores both the theoretical and practical dimensions of decentralised international cooperation. This concept is framed in existing international environmental law principles and regimes, and is used as an interpretative approach to provide an innovative and bottom-up reading of international environmental law. I argue that this concept acknowledges and legitimises the role of local actors at the international level and has implications in terms of effective participation, benefit-sharing, and environmental governance more generally. Four case studies are used to show how this concept has been operationalised in the European and southern African contexts: respectively, two European Groupings of Territorial Cooperation (EGTCs) – the ZASNET and Alpi Marittime-Mercantour – and two Transfrontier Conservation Areas (TFCAs) – the Kavango Zambezi and Great Limpopo. A comparative analysis of these cases provides a useful approach not only for understanding and potentially strengthening existing instances of decentralised international cooperation, but also for facilitating the developement of such cooperation in other contexts. This comparison revealed a number of trends (legal harmonisation, stretching of normative boundaries, etc.) that may be useful in improving biodiversity conservation. Moreover, the availability of institutional mechanisms appears to affect how quickly and efficiently participation of local actors can take place. The main strengths of decentralised international cooperation are that it promotes the transboundary dimension of nature conservation and the active involvement of sub-national actors. By so doing, it bridges between governance levels and can contribute to shaping a more appropriate and participatory framework for the governance of shared natural resources.

This thesis aimed to explore eukaryotic cellular processes upon the virulent attack of low doses of a well-known pore forming toxin (staphylococcal α-hemolysin (αHL)) and to develop a new biotech application using the same protein.

Post-conflict situations, regime changes, and peace processes within situations of active hostility pose tremendous challenges for the governments and societies involved. The inherent dilemma between justice and the international obligations to investigate and prosecute a legacy of large-scale human rights abuses, and the role of alternative justice mechanisms, which seek to pursue more abstract goals such as peace and reconciliation, has its roots in the current discourse of transitional justice. By analyzing the provisions in which the obligations to investigate and prosecute are enshrined or derived, this work disentangles the common misconception that such procedural obligations are naturally rooted and clearly spelled out in the most important international humanitarian and human rights instruments considered by this study. It also dispels the notion that successor governments facing a transition operate in a normative vacuum: international law, human rights bodies, the ICC, and the UN impose limitations on the sovereign exercise of states’ prerogatives, thus shaping local approaches to policies of justice. But an uncompromising call for prosecution overlooks realpolitik considerations, practical difficulties, and the peculiar needs of fragile successor governments with limited power and room for maneuver. Granted that the rule of law cannot be reestablished in a society in which perpetrators enjoy impunity, since de facto and de jure impunity do per se constitute a breach of states parties’ duties under the conventions, criminal prosecution cannot be the only tool for reckoning with the past because punishment also encompasses non-criminal sanctions. A pluralistic notion of accountability and an integrated approach to peacemaking are thus advocated. International law is only capable of adjusting to the peculiarities of transitions if it is flexibly understood and applied. To remain relevant as a legal regime it needs to accord with political realities but should always be interpreted in a manner consistent with its rationale.

Atrial fibrillation (AF) is the most common sustained arrhythmia worldwide and a frequent cause of hospitalization. Moreover, it represents one of the most frequent complication following cardiac surgery with an incidence of around 30% and an important predictor of patient morbidity. The exact pathophysiological mechanisms responsible for the onset and perpetuation of AF are not completely understood. However, clinical and experimental insights on the factors causing AF have suggested that atrial fibrillation is a multi-factorial phenomenon. Atrial fibrillation is characterized by a highly complex and irregular electrical activation of the atrial tissue, which is the manifestation of diverse abnormalities (electrical, structural, metabolic, neurohormonal, and/or molecular alterations) in diverse pathological conditions. In particular, it has been shown that fibrosis, a phenomenon in which extracellular matrix (ECM) components, mainly fibrillar collagen, accumulate between cardiomyocytes, leads to the inhomogeneous atrial electrical conduction typical of fibrillation. Recent studies have suggested that the deregulation of gene expression may act as a molecular mechanism of arrhythmogenesis. In particular, miRNAs, a new class of non-coding RNAs have rapidly emerged as one of the key players in the gene expression regulatory network, so variations in their expression levels may constitute a pathway for the arrhythmia-induced atrial remodeling. The present study aims to investigate the structural and molecular features of atrial tissue, with particular attention to fibrosis, which may be involved in the formation of a pro-arrhythmic substrate. By using both histological and advanced microscopy techniques, intramural fibrotic content and 3D collagen network properties were determined in atrial samples, collected during cardiac surgery in patients who developed or not AF. The quantitative analysis indicated a general decrease of collagen content from the outer (the epicardium) to the inner (the endocardium) myocardial wall, in the overall patient population. However, AF patients presented higher fibrotic values compared to sinus rhythm (SR) patients in the deeper myocardial layers, thus supporting the hypothesis that an accumulation of fibrotic tissue within the myocardial wall may represent an important structural contributor in the pathophysiology of AF. In addition to a quantitative assessment, collagen properties such as fibers orientation (degree and anisotropy) and scale dimension, were determined by non-linear optical microscopy techniques. The analysis revealed that in SR patients collagen network showed a fine architecture characterized by thin fibrils with changing angles and directions compared to AF, where fibers tended to pack-up in larger bundles of defined directions. A quantitative analysis of the 3D collagen network features, throughout the atrial wall, revealed that fibers orientation and scale dimension changed along tissue depth in both SR and AF patients, with larger values of orientation and fiber changes in AF tissues. These results highlight the spatial rearrangement and thickening of the 3D collagen network in AF patients, suggesting its possible role in the maintenance of the arrhythmia. Numerous evidence indicated that also an altered regulation of gene expression may play an important role in the mechanisms of atrial remodeling which underlie AF. In this perspective, the expression pattern of some miRNAs known to target different genes involved in diverse mechanisms that underlie AF was evaluated. A panel of miRNAs (miR-1, miR-133a/b, miR-30c, miR-29a/b, miR-208a/b, miR-328, miR-499, miR-590 and miR-21), principally involved in the formation of a pro-arrhythmic substrate, was selected after an accurate review of the literature and analyzed by RT-qPCR, in AF patients versus SR individuals. To accurately determine the levels of analyzed miRNAs, their expression data are usually normalized relatively to endogenous and/or exogenous reference genes. To date, no general agreement between different normalization strategies has been found, in particular in cardiac tissue, for the study of AF. For these reasons, a preliminary study aiming to establish the best endogenous reference genes for miRNAs data normalization was performed. Specifically, different well-established analysis tools such as NormFinder, GeNorm, BestKeeper and ∆Ct method, were applied on five commonly used endogenous reference transcripts such as 5S, U6, SNORD48, SNORD44 and miR-16. The suitable reference gene obtained, SNORD48, was applied for miRNAs data normalization. Our findings revealed that miRNAs expression levels were different in AF compared to SR patients. MiR-208a and miR-208b displayed statistically significant differences between the two populations. To investigate possible relationships between miRNAs expression levels and the fibrotic content a correlation measurement was also performed. Our analysis revealed that miR-21 and miR-208b were close to a significant correlation with fibrosis. In conclusion, this work introduced new techniques and implemented new methods of analysis for the study of the substrate of AF. In particular, the results obtained with this multiscale approach, from structural to molecular level, exacerbated the role of fibrosis as a critical contributor in the formation of a pro-arrhythmic substrate. Nonetheless, further studies are needed for a better understanding of the ways in which structural, molecular and also cellular remodeling may alter the impulse propagation in the myocardium.

The DPRK is the only State that attempted to leave the Nuclear Non-Proliferation Treaty (NPT) in 1993 and 2003. In accordance with Article X(1) of the NPT, a withdrawing State is required, inter alia, to submit notifications of withdrawal to all States Parties to the NPT and to the United Nations Security Council (UNSC). However, the role of the UNSC in such a case is not defined in Article X(1) of the NPT. Thus, both in 1993 and 2003, there were disagreements among the UNSC members on the involvement of the UNSC into the matter and its possible actions to respond to the announcements of withdrawal from the NPT. The UNSC was criticized for either not fully deploying its mandate under the UN Charter, or intervening in the matter of withdrawal. In some cases, actions of the UNSC were regarded as an infringement of the sovereign right of States to leave Treaties. This PhD dissertation assesses the powers of the UNSC under the UN Charter that gives the UNSC the mandate to take actions in case of threat to international peace and security. The dissertation focuses on Article 39 of the UN Charter, under which the UNSC defines such threat; and on the competence of the UNSC to pass binding decisions under Chapter VII of the UN Charter. The dissertation concludes that the UNSC has the authority to define withdrawal from the NPT as a threat to international peace and the security and consequently to take actions under Chapter VII.

Structural optimization has become an important tool for structural designers, since it allows a better exploitation of material, thus decreasing structure self-weight and saving material costs. Moreover, it helps the designer to find innovative design solutions and structural forms that not only better exploit material but also give the structure higher aesthetic value from an architectural point of view. When applied to real scale structures like bridges, this approach leads to the definition of voids patterns delimiting regions where fluxes of force migrate from force application point to boundary regions and suggests innovative layouts without renouncing to formal and structural aspects. Nevertheless, the criticality of this powerful tool is related to the ease of defining entire families of possible candidate solutions, by modifying input volume reduction ratio to reduce structural weight as much as possible or defining several starting trial solutions based on the judgment of designer. In this case, structural optimization still leads to the best material distribution, but finding the best compromise between material saving and structural performance is a designer choice. To face this aspect, a global optimization index (GOI) has been defined and applied to the structural optimization of a steel-concrete arch bridge built is San Donà in the province of Venice, Italy. On the basis of this work, a generalized version of the optimization index is proposed and its analytical formulation is discussed in detail in this thesis. The application of proposed optimization index is extended from topology optimization to other optimization techniques. Moreover it allows not only to identify best candidate solution originated by a unique reference model, but even comparing structural performances between candidates solution derived by several starting trial solutions. Through structural optimization procedure performed on three different type bridges, namely footbridges supported by concrete shell, Calatrava Bridge (steel arch bridge) and two cable-stayed bridges, the effectiveness of proposed optimization index is validated. The results show that the proposed optimization index provides to the designer a mathematical procedure able to highlight the best choice among several candidate solutions obtained by the optimization procedure. With the proposed optimization index, a suitable score for each design solution of specific starting layout is assigned, therefore the best overall layout solution which is the best compromise between material saving and structural performance can be highlighted among single-family multi-solutions or multi-families or multi-solutions.

The students of the Self-Reliant Learning Program of the Halton Board of Education participate in an alternative secondary school program. They work on individualized, student-centered programs and proceed at a self-determined rate. The students range in age from 17 to 70 years, have a greater sense of their educational needs and direction and may bring to the classroom wider experiences than the students of a regular secondary school. A problem-based course has been developed to assist the students in developing skills for dealing with problems while gaining knowledge of biology. The limited field testing that has been carried out indicates that the students in the course are improving their organizational and problem-solving skills and showing improved understanding of biological concepts. The educational rationale and an evaluation of the program are presented along with the materials comprising the seven units of the course. / Thesis / Master of Science (Teaching)

Background and rationale Translation is a fundamental biological process occurring in cells, carried out by ribosomes simultaneously bound to an mRNA molecule (polyribosomes). It has been exhaustively demonstrated that dysregulation of translation is implicated in a wide collection of pathologies including tumours and neurological disorders. Latest findings reveal the existence of translational regulatory mechanisms acting in cis or trans with respect to the mRNAs and governing the movement and the position of ribosomes along transcripts or directly impacting on the ribosome catalogue of its constituent proteins. For this reason, translational controls also account for widespread uncoupling between transcript and protein abundances in cells. To explain the poor correlation between transcripts and protein levels, many computational models of translation have been developed. Usually, these approaches aim at predicting protein abundances in cells starting from the mRNA abundance. Despite the efforts of these modelling studies, a consensus model remains elusive, drawing to contradictory conclusions concerning the role of mRNA regulatory elements such as the usage of codons (codon usage bias) and slowdown mechanism at the beginning of the coding sequence (ramp). More recently, following the rapid and widespread diffusion of ribosome footprinting assays (RiboSeq), which enables the dissection of translation at single nucleotide resolution, a number of computational pipelines dedicated to the analysis of RiboSeq data have been proposed. These tools are typically designed for extracting gene expression alterations at the translational level, while the positional information describing fluxes and positions of ribosomes along the transcript is still underutilized. Therefore, the polysome organization, in term of number and position of ribosomes along the transcript and the translational controls directed in shaping cellular phenotypes is still open to breakthrough discoveries. Broad objectives The aim of my thesis is the development of mathematical and computational tools integrated with experimental data for a comprehensive understanding of translation regulation and polysome organization rules governing the number of ribosomes per polysome and the ribosome position along transcripts. Project design and methods With this purpose, I developed riboWaves, an integrated bioinformatics suite divided in two branches. riboWaves includes in the first branch two modeling modules: riboAbacus, predicting the number of ribosomes per transcript, and riboSim, predicting ribosome localization along mRNAs. In the second branch, riboWaves provides two pipelines, riboWaltz and riboScan, for detailed analyses of ribosome profiling data aimed at providing meaningful and yet unexplored ribosome positional information. The models and the pipelines are implemented in C and R, respectively. riboAbacus and riboWaltz are available on GitHub. Results To predict the number of ribosomes per transcript and the position of ribosomes on mRNAs, I applied riboAbacus and riboSim, respectively, to transcriptomes of different organisms (yeast, mouse, human) for understanding the role of translational regulatory elements in tuning polysome in different organisms. First, I trained and validated performances of riboAbacus taking advantage of Atomic Force Microscopy images of polysomes, while performances of riboSim were assessed employing ribosome profiling data. Predictions provided by riboAbacus and riboSim were evaluated in parallel. I showed that the average number of ribosomes translating a molecule of mRNA can be well explained by the deterministic model, riboAbacus, that includes as features the mRNA levels, the mRNA sequences, the codon usage bias and a slowdown mechanism at the beginning of the CDS (ramp hypothesis). The predictions of ribosome localization by riboSim that used as features the mRNA sequence, the codon usage and the ramp, were run for yeast, mouse and human. I observed a good similarity between the predicted and experimental positions of ribosomes along transcripts in yeast, while poor similarity was obtained between predicted and experimental ribosome positions in the two mammals, suggesting the presence of more elaborate controls that tune ribosomes movement in higher eukaryotes than in simple species. After having developed two tools for the analyses of RiboSeq data and extraction of positional information on ribosome localization along transcripts, I applied both riboWaltz and riboScan in a case study. The aim was to dissect possible defects in ribosome localization in tissues of a mouse model of Spinal Muscular Atrophy (SMA). SMA is a neurodegenerative disorder caused by low levels of the Survival of Motor Neuron protein (SMN) in which translational impairments are recently emerging as possible cause of the disease. I analysed ribosome profiling data obtained from three different types of RiboSeq variants in healthy and SMA-affected mouse brains at the early-symptomatic stage of the disease. I observed i) a significant drop-off of translating ribosomes along the coding sequence in the SMA condition (using riboWaltz); ii) in SMA-affected mice, the possible accumulation of ribosomes along the 3' UTR in neuro-related mRNAs (using riboScan); iii) the involvement of SMN-specialized ribosomes in playing a very intimate role with the elongation stage of translation of the first codons of transcripts (riboWaltz), iv) the loss of ribosomes at the 3rd codon in SMA in transcripts bound by SMN-specialized ribosomes and v) a remarkable connection between SMN and the down-regulation of genes in SMA-affected mice. Overall, these findings confirmed previous observation about possible SMN-related dysregulations of local protein synthesis in neurons. More importantly, they unravel a completely new role of SMN in tuning translation at multiple levels (initiation, elongation and the recycling of terminating ribosomes), opening new hypotheses and scenarios for explaining the most devastating genetic disease, leading cause worldwide of infant mortality. Conclusions The present work provides a new comprehensive and integrated scenario for better understanding translation and demonstrates that this approach is a very powerful strategy to pave the way for new understanding of fine alteration in polysome organization and functional control in both physiological and pathological conditions.

The duty not to betray the social group is an ancient socio-political concept adopted by authorities to regulate the loyal behaviour of their members. Such duty has been legally regulated for centuries and now forms part of the domestic legal systems of contemporary states. Known in English as allegiance, the duty of fidelity is a fundamental concept of law that gains notoriety in times of armed conflicts. However, allegiance has been overlooked by scholars of international law as a factor in the design, interpretation and application of the rules of international humanitarian law (IHL) that protect individuals during armed conflicts and limit the capacity of states to choose methods of warfare. This study analyses the role that the concept of allegiance plays in the rules of IHL applicable in international armed conflicts. This is done by determining the contemporary meaning of allegiance in state practice, the ways it has been introduced into international law, and its use by relevant actors in six areas of IHL. The investigation reveals a rich historical practice around the concept of allegiance, its incorporation in rules and discussions of IHL, and the employment of this concept for multiple and diverse purposes: from the alleged basis for conferring protected status to civilians, to the alleged basis for denying protected status to combatants; from a protected element of occupied population, to a corruptible element of enemy population outside occupation; and as a presumption of dangerousness for enemy aliens, the required mens rea for desertion, and a vague term to determine connection between belligerent parties and armed groups. This study clarifies the contemporary understanding of allegiance, and confronts the concept with treaty provisions, case law, and academic debates relevant to the use of allegiance in IHL. Additionally, the study explores the relation between domestic law and international law applicable during international armed conflicts and the overlooked position of individuals of dubious loyalties such as traitors, collaborators, deserters, internal enemies, and corrupters.

The provision of relief to civilians in armed conflict is a sensitive activity, subject to specific regulation in IHL treaties. Challenges emerged on the ground have questioned the comprehensive nature of this legal framework and generated debate on the concept of humanitarian assistance itself, the role of different kinds of actors (local/external, governmental/nongovernmental, armed/unarmed) in providing it, and the value and meaning of the principles traditionally associated to it—humanity, impartiality, neutrality, independence. This research, examining the evolution of State practice and opinio juris, provides a comprehensive analysis of the legal regime applicable to the provision of relief to civilians in armed conflict and the different categories of actors involved in it, identifying answers offered by international law (primarily IHL) to issues emerged in practice. It is argued that humanitarian assistance is a well-defined and limited concept under IHL. Rules on this issue have been subject to progressive development, e.g. those on the protection of humanitarian workers in non-international armed conflict, but State practice has revealed that sovereignty remains important, and the principles of humanitarian assistance continue to embody the balance acceptable to States between military necessity and humanitarian considerations. No right to access or to provide humanitarian assistance without consent from the Parties concerned has developed, including no right to provide relief in non-international armed conflict in territory controlled by non-State armed groups without State consent. Participation in the provision of humanitarian assistance by local and external actors is not prohibited, but the level of protection they enjoy depends on their position under IHL. Different regimes are applicable to distinct armed actors (belligerents/peacekeepers/external armed forces/private security companies) but in all cases respect for the principle of distinction is central. In general, special protection for relief actions and actors remains connected to respect for the principles of humanitarian assistance. This has been confirmed by belligerents’ reactions to the increased engagement of humanitarian organisations in protection, as the second essential component of humanitarian action: belligerents have claimed their entitlement to require respect by humanitarian personnel also for the most contested principle—neutrality, meaning non-interference in hostilities and even abstention from involvement in politics.