Systematic Analysis of FKBP Inducible Degradation Domain Tagging Strategies for the Human Malaria Parasite Plasmodium falciparum

Targeted regulation of protein levels is an important tool to gain insights into the role of proteins essential to cell function and development. In recent years, a method based on mutated forms of the human FKBP12 has been established and used to great effect in various cell types to explore protein function. The mutated FKBP protein, referred to as destabilization domain (DD) tag when fused with a native protein at the N- or C-terminus targets the protein for proteosomal degradation. Regulated expression is achieved via addition of a compound, Shld-1, that stabilizes the protein and prevents degradation. A limited number of studies have used this system to provide powerful insight into protein function in the human malaria parasite Plasmodium falciparum. In order to better understand the DD inducible system in P. falciparum, we studied the effect of Shld-1 on parasite growth, demonstrating that although development is not impaired, it is delayed, requiring the appropriate controls for phenotype interpretation. We explored the quantified regulation of reporter Green Fluorescent Protein (GFP) and luciferase constructs fused to three DD variants in parasite cells either via transient or stable transfection. The regulation obtained with the original FKBP derived DD domain was compared to two triple mutants DD24 and DD29, which had been described to provide better regulation for C-terminal tagging in other cell types. When cloned to the C-terminal of reporter proteins, DD24 provided the strongest regulation allowing reporter activity to be reduced to lower levels than DD and to restore the activity of stabilised proteins to higher levels than DD29. Importantly, DD24 has not previously been applied to regulate proteins in P. falciparum. The possibility of regulating an exported protein was addressed by targeting the Ring-Infected Erythrocyte Surface Antigen (RESA) at its C-terminus. The tagged protein demonstrated an important modulation of its expression.

An assessment of the business value of traceability practices in the italian fishery processing industry

Traceability is often perceived by food industry executives as an additional cost of doing business, one to be avoided if possible. However, a traceability system can in fact comply the regulatory requirements, increase food safety and recall performance, improving marketing performances and, as well as, improving supply chain management. Thus, traceability affects business performances of firms in terms of costs and benefits determined by traceability practices. Costs and benefits affect factors such as, firms’ characteristics, level of traceability and ,lastly, costs and benefits perceived prior to traceability implementation. This thesis was undertaken to understand how these factors are linked to affect the outcome of costs and benefits. Analysis of the results of a plant level survey of the Italian ichthyic processing industry revealed that processors generally adopt various level of traceability while government support appears to increase the level of traceability and the expectations and actual costs and benefits. None of the firms’ characteristics, with the exception of government support, influences costs and level of traceability. Only size of firms and level of QMS certifications are linked with benefits while precision of traceability increases benefits without affecting costs. Finally, traceability practices appear due to the request from “external“ stakeholders such as government, authority and customers rather than “internal” factors (e.g. improving the firm management) while the traceability system does not provide any added value from the market in terms of price premium or market share increase.

The Council of Ministers of the European Union After the Lisbon Treaty

The Treaty of Lisbon has brought remarkable changes and innovations to the European Union. As far as the Council of Ministers of the European Union (“the Council” hereinafter) is concerned, there are two significant innovations: double qualified majority voting and new rotating Presidency scheme, which are considered to make the working of the Council more efficiently, stably and consistently. With the modification relating to other key institutions, the Commission and the European Parliament, and with certain procedures being re-codified, the power of the Council varies accordingly, where the inter-institutional balance counts for more research. As the Council is one of the co-legislatures of the Union, the legislative function of it would be probably influenced, positively or negatively, by the internal innovations and the inter-institutional re-balance. Has the legislative function of the Council been reinforced or not? How could the Council better reach its functional goal designed by the Treaties’ drafter? How to evaluate the Council’s evolution after Lisbon Treaty in the light of European integration? This thesis is attempting to find the answers by analyzing two main internal innovations and inter-institutional re-balance thereinafter.

Welfare Lobby Groups responding to Globalisation: A Comparison of the Australian Council of Social Service (ACOSS) and the UK Child Poverty Action Group (CPAG)

The past decade has witnessed a period of intense economic globalisation. The growing significance of international trade, investment, production and financial flows appears to be curtailing the autonomy of individual nation states. In particular, globalisation appears to be encouraging, if not demanding, a decline in social spending and standards. However, many authors believe that this thesis ignores the continued impact of national political and ideological pressures and lobby groups on policy outcomes. In particular, it has been argued that national welfare consumer and provider groups remain influential defenders of the welfare state. For example, US aged care groups are considered to be particularly effective defenders of social security pensions. According to this argument, governments engaged in welfare retrenchment may experience considerable electoral backlash (Pierson 1996; Mishra 1999). Yet, it is also noted that governments can take action to reduce the impact of such groups by reducing their funding, and their access to policy-making and consultation processes. These actions are then justified on the basis of removing potential obstacles to economic competitiveness (Pierson 1994; Melville 1999).

CAP Reform vs WTO: Crop Diversification or Crop Rotation? : Legal opinion prepared by Dr. Christian Häberli at the request of APRODEV, the Association of World Council of Churches related Development Organisations in Europe

The European Commission’s proposals for the Legislative Framework of the Common Agricultural Policy (CAP) in the period 2014-2020 include, inter alia, the introduction of a “strong greening component”. For the first time, all EU farmers in receipt of support are to “go beyond the requirements of cross compliance and deliver environmental and climate benefits as part of their everyday activities crop diversification as a contribution to all EU farmers in receipt of support go beyond the requirements of cross compliance and deliver environmental and climate benefits as part of their everyday activities.” In a legal opinion prepared at the request of APRODEV, the Association of World Council of Churches related Development Organisations in Europe (www.aprodev.eu), Christian Häberli examines the WTO implications of this proposal, as compared with an alternative proposal to rather link direct payments to crop rotation. The conclusions are twofold: 1. Crop rotation is at least as likely to be found Green Box-compatible as crop diversification. Moreover, it will be more difficult to argue that crop diversification is “not more than minimally production-distorting” because it entails for most farmers less cost and work. 2. Even if (either of the two cropping schemes) were to be found “amber”, the EU would not have to relinquish this conditionality. This is because the direct payments involved would in all likelihood not, together with the other price support instruments, exceed the amount available under the presently scheduled maximum.