Qualitative and absolute quantitative studies of the cell-nanoparticle interaction

This thesis focused on the polymer’s influence on the interaction of polymeric NPs with epithelial cells. Furthermore, the measurement of single submicron nanoparticles in a commercially available flow cytometer was established, to provide a new method in the toolbox for nanoparticle-cell studies. This gave way to develop a routine for the absolute quantification of intracellular NPs via flow cytometry. rnThe cellular uptake of poly(methyl methacrylate) (PMMA), polystyrene (PS) and poly(L-lactide) (PLLA) nanoparticles was investigated via flow cytometry. PLLA-NPs were internalized the most efficiently. But upon co-incubation of PS and PLLA particles with cells, the two particles mutually influenced their uptake, slightly shifting the relative uptake efficiencies. This phenomenon should be based on specific properties of the different polymer materials. The findings indicated a competition (which is strongly influenced by properties of the respective polymeric material) for the uptake into the cells, allegedly due to competition for specific coatings with serum components that enhances the NPs’ cellular uptake. The fluorescence of single 150 nm particles was determined with a benchtop cytometer, breaching the machine’s detection limit but yielding precise NP fluorescence standardization factors. Up to now, these standardization factors are mostly determined by spectroscopic analysis of the particles’ dye content. Finally a flow cytometric routine for absolute particle counting in cells was devised. This quantitation revealed a low uptake efficiency for un-functionalized PMMA NPs of less than 150 NPs (approx. 0,001 % of added) per cell.rn

Atropisomeric xanthines: Synthesis, stereodynamics and absolute configuration

During the thesis period a new class of atropisomeric xanthine derivatives has been studied. We decided to focus our attention on these purine bases because of their various biological activities, that could play an important role in the discovery of new bioactive atropisomers. The synthesized compounds bear an Aryl-N chiral axis in position 1 of the xanthine scaffold, around which the rotation is prevented by the presence of bulky ortho substituents. Through a retro synthetic analysis we synthesized three atropisomeric structures bearing in position 1 of the purine scaffold respectively an o-tolyl, o-nitrophenyl and a 1-naphthyl group. The conformational studies by DFT simulations showed that the interconversion energy barrier between the two available skewed conformations is higher enough to obtain thermally stable atropisomers. After the separation of the atropisomers, the experimental energy of interconversion was investigated by means of kinetic studies following the thermal racemization process using an enantioselective HPLC column. The absolute configuration of each atropisomer was assigned by experimental ECD analysis and TD-DFT simulations of the ECD spectra.

Remaining lifetime and absolute 10-year probabilities of osteoporotic fracture in Swiss men and women

SUMMARY: Remaining lifetime and absolute 10-year probabilities for osteoporotic fractures were determined by gender, age, and BMD values. Remaining lifetime probability at age 50 years was 20.2% in men and 51.3% in women and increased with advancing age and decreasing BMD. The study validates the elements required to populate a Swiss-specific FRAX model. INTRODUCTION: Switzerland belongs to high-risk countries for osteoporosis. Based on demographic projections, burden will still increase. We assessed remaining lifetime and absolute 10-year probabilities for osteoporotic fractures by gender, age and BMD in order to populate FRAX algorithm for Switzerland. METHODS: Osteoporotic fracture incidence was determined from national epidemiological data for hospitalised fractured patients from the Swiss Federal Office of Statistics in 2000 and results of a prospective Swiss cohort with almost 5,000 fractured patients in 2006. Validated BMD-associated fracture risk was used together with national death incidence and risk tables to determine remaining lifetime and absolute 10-year fracture probabilities for hip and major osteoporotic (hip, spine, distal radius, proximal humerus) fractures. RESULTS: Major osteoporotic fractures incidence was 773 and 2,078 per 100,000 men and women aged 50 and older. Corresponding remaining lifetime probabilities at age 50 were 20.2% and 51.3%. Hospitalisation for clinical spine, distal radius, and proximal humerus fractures reached 25%, 30% and 50%, respectively. Absolute 10-year probability of osteoporotic fracture increased with advancing age and decreasing BMD and was higher in women than in men. CONCLUSION: This study validates the elements required to populate a Swiss-specific FRAX model, a country at highest risk for osteoporotic fractures.

Strict and non-strict independent and-parallelism in logic programs: Correctness, efficiency, and compile-time conditions.

This paper presents some fundamental properties of independent and-parallelism and extends its applicability by enlarging the class of goals eligible for parallel execution. A simple model of (independent) and-parallel execution is proposed and issues of correctness and efficiency discussed in the light of this model. Two conditions, "strict" and "non-strict" independence, are defined and then proved sufficient to ensure correctness and efñciency of parallel execution: if goals which meet these conditions are executed in parallel the solutions obtained are the same as those produced by standard sequential execution. Also, in absence of failure, the parallel proof procedure does not genérate any additional work (with respect to standard SLD-resolution) while the actual execution time is reduced. Finally, in case of failure of any of the goals no slow down will occur. For strict independence the results are shown to hold independently of whether the parallel goals execute in the same environment or in sepárate environments. In addition, a formal basis is given for the automatic compile-time generation of independent and-parallelism: compile-time conditions to efficiently check goal independence at run-time are proposed and proved sufficient. Also, rules are given for constructing simpler conditions if information regarding the binding context of the goals to be executed in parallel is available to the compiler.