Role of Magnesium and its mitochondrial transporter MRS2 in the modulation of drug-induced apoptosis leading to multidrug resistance phenotype

Magnesium is an essential element for many biological processes crucial for cell life and proliferation. Growing evidences point out a role for this cation in the apoptotic process and in developing multi drug resistance (MDR) phenotype. The first part of this study aimed to highlight the involvement of the mitochondrial magnesium channel MRS2 in modulating drug-induced apoptosis. We generated an appropriate transgenic cellular system to regulate expression of MRS2 protein. The cells were then exposed to two different apoptotic agents commonly used in chemotherapy. The obtained results showed that cells overexpressing MRS2 channel are less responsiveness to pharmacological insults, looking more resistant to the induced apoptosis. Moreover, in normal condition, MRS2 overexpression induces higher magnesium uptake into isolated mitochondria respect to control cells correlating with an increment of total intracellular magnesium concentration. In the second part of this research we investigated whether magnesium intracellular content and compartmentalization could be used as a signature to discriminate MDR tumour cells from their sensitive counterparts. As MDR model we choose colon carcinoma cell line sensitive and resistant to doxorubicin. We exploited a standard-less approach providing a complete characterization of whole single-cells by combining X-Ray Fluorescence Microscopy , Atomic Force Microscopy and Scanning Transmission X-ray Microscopy. This method allows the quantification of the intracellular spatial distribution and total concentration of magnesium in whole dehydrated cells. The measurements, carried out in 27 single cells, revealed a different magnesium pattern for both concentration and distribution of the element in the two cellular strains. These results were then confirmed by quantifying the total amount of intracellular magnesium in a large populations of cells by using DCHQ5 probe and traditional fluorimetric technique.

La trasformazione dei processi legislativi nella società dell'informazione: dall'e-Legislation all'e-Parliament

The transformation of legislative processes in the Information society: from eLegislation to eParliament This research analyzes, by means of an interdisciplinary and comparative approach, the transformation of legislative processes produced by the introduction of new ICT technologies. The use of ICT in support of parliamentary activities is concerned with efficiency of parliamentary process and aims at more transparent procedures, improved access to documents, social participation and cooperation among institutions. With ICT Parliaments are now able to improve their efficiency and optimize their business; they can advance the dialogue with their citizen both, through the real access and the effective availability of information and, through new way of participation in the democratic process. Finally, sharing information, know-out, best practices and other records, Parliaments will be able to develop new information and knowledge and to strengthen the role and power of Institutions. Only through a global vision of the full process, re-thinking and develop rules and uniform standard and so implementing the new opportunities carrying out by ICT, it will be possible to put in practice concrete eParliament results. The Research goals are at least three: 1. To Analysed the legislative process and the ICT opportunities to understand the impact of the latter on the former. In particular to check up the problems that ICT can raise in relation of the constitutional principles ensuring the process itself. 2. To realized an abstract model representing the legislative process regardless of the form of government, chambers composition, legal system, etc. 3. To suggest standard, structural, linguistic and ontological, able to implement the new opportunities of sharing, cooperation and reuse among the many and various stakeholders of the democratic/legislative view.

Rebooting the constitutional debate: deliberative constitutionalism in the European Union

The aim of my thesis is to investigate the possibility and necessity to rethink a constitutional framework and debate in a transnational polity such as the European Union. My effort focuses on a promising theory called deliberative constitutionalism, which carries on new insights on how democracy and constitutions relate each other. The EU is a unique political entity which poses unanswered questions about its political legitimacy and constitutional foundation, if a Constitution will ever be possible. Going beyond the classical conception of the national and sovereign ‘people’, we keep wondering how citizens may deliberate and discuss about their rights and political communities across borders, in what could be defined as a transnational civic society. The development of the latter brings with it necessary constitutional changes, if not an evolution of constitutionalism itself. Chapter 1 deals with defining the theoretical framework, which develops the distinctiveness of the deliberative constitutional paradigm not only with respect to other more 'classical' models of democracy, but also with respect to other deliberative models that have marked the constructivist debate. Chapter 2 presents a conceptual history of constituent power, mainly studying the evolution of the constitution-sovereignty-constituent power dialectic, up to contemporary theories that explain the negation, separation, union or plurality of a transnational constituent with respect to its national counterparts. Chapter 3 develops the discourse of constitutional pluralism, through its main claims and strands that especially pertain to Neil Walker's (2002, 2016) institutional and epistemic claims. Chapter 4 applies a deliberative constitutionalist framework to the case of the European Union. Through the exposition of DC normative tenets, a form of self-learning process is proposed that can reconcile the heterarchical arrangement of constitutional claims and the new demand for legitimacy, as well as the relationship between European peoples and European citizens.