Stabilization of troponin I for applications in clinical chemistry and forensic medicine. Cardiac troponin I: A time of death marker

Cardiac troponin I (cTnI) is one of the most useful serum marker test for the determination of myocardial infarction (MI). The first commercial assay of cTnI was released for medical use in the United States and Europe in 1995. It is useful in determining if the source of chest pains, whose etiology may be unknown, is cardiac related. Cardiac TnI is released into the bloodstream following myocardial necrosis (cardiac cell death) as a result of an infarct (heart attack). In this research project the utility of cardiac troponin I as a potential marker for the determination of time of death is investigated. The approach of this research is not to investigate cTnI degradation in serum/plasma, but to investigate the proteolytic breakdown of this protein in heart tissue postmortem. If our hypothesis is correct, cTnI might show a distinctive temporal degradation profile after death. This temporal profile may have potential as a time of death marker in forensic medicine. The field of time of death markers has lagged behind the great advances in technology since the late 1850's. Today medical examiners are using rudimentary time of death markers that offer limited reliability in the medico-legal arena. Cardiac TnI must be stabilized in order to avoid further degradation by proteases in the extraction process. Chemically derivatized magnetic microparticles were covalently linked to anti-cTnI monoclonal antibodies. A charge capture approach was also used to eliminate the antibody from the magnetic microparticles given the negative charge on the microparticles. The magnetic microparticles were used to extract cTnI from heart tissue homogenate for further bio-analysis. Cardiac TnI was eluted from the beads with a buffer and analyzed. This technique exploits banding pattern on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by a western blot transfer to polyvinylidene fluoride (PVDF) paper for probing with anti-cTnI monoclonal antibodies. Bovine hearts were used as a model to establish the relationship of time of death and concentration/band-pattern given its homology to human cardiac TnI. The final concept feasibility was tested with human heart samples from cadavers with known time of death. ^

Drosophila Flight Stabilization Depends on Apparent Distances in Optic Flow Field

To perform daily flight tasks, insects rely heavily on their visual perception of a dynamic environment. They must process visual signals quickly and accurately and update their behavior. Flies are vulnerable to environmental disturbances, such as gusts of wind blowing them off course, but they may use the altered visual field to compensate and regain their original course. In studies using Drosophila melanogaster, it has been shown that their corrective responses can be analyzed by measuring changes in their wing beats. By enclosing a tethered fly in a cuboidal visual arena displaying a computerized optic flow field, it is possible to calculate the change in wing beat amplitudes from an infrared shadow of its wings using photodiodes and a custom wing beat analyzer. In this experiment, manipulations ofthe optic flow field are used to create a field where points have varying relative forward speed, to study how the insect performs corrective maneuvers. The results show that Drosophila have a stronger corrective response to the quickly moving, apparently near points compared to the slower moving, apparently distant points. This implies the flies are distinguishing points based on their relative speeds, inferring distance, and adjusting their corrective actions with this information.

Democratization, stabilization and social movements : the Bolivian case

In this thesis I assessed the state responses to social movements and in turn the impact of these movements on state policy within the context of the democratization occurring in Bolivia. The democratization process is affected by the conflict between political and economic goals. Politically the governments are faced with the demands from social groups. At the same time, the Bolivian government faces an economic crisis which requires stabilization, impairing the same individuals needed for legitimacy and political support. Two cases which depicted the key issues of this thesis are: the indigenous groups in the Bolivian Beni region and the coca growers, mainly of the Chapare area in the Cochabamba department of Bolivia. To achieve support and legitimacy, the new civilian administrations had no choice but to listen to the requests of the social mobilizations. Because of the economic crisis, conflicting domestic pressures and international influence, however, the government could not accede to all their demands.

The Central European initiative : an approach to regional stabilization and democratic consolidation

This thesis examines how multilateral institutions can contribute to democratization and regional stability. It is a case study of the Central European Initiative (CEI), a regional intergovernmental organization founded by Italy and Austria in 1989 to undertake a regional integration process, and of its role in stabilizing post-communist democracies. Documents were collected at the CEI offices in Trieste, Italy, and interviews conducted with CEI officers, and data obtained from the websites of related organizations and of CEI member countries. The thesis probes the relevance of concepts derived from theories of international regimes and social constructivism. It shows that the CEI diffuses norms and institutional rules conducive to consolidating democracies, including the development of a free civil society, a relatively autonomous political society, rule of law, state bureaucracies that are usable by the new democratic governments, and functioning free market economies, addition, and fosters habits of dialogue, socializes participants, and supports the creation of supra-national identities.