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    Recent developments in peptide-based nucleic acid delivery
    (Basel : MDPI, 2008) Veldhoen, Sandra; Laufer, Sandra D.; Restle, Tobias
    Despite the fact that non-viral nucleic acid delivery systems are generally considered to be less efficient than viral vectors, they have gained much interest in recent years due to their superior safety profile compared to their viral counterpart. Among these synthetic vectors are cationic polymers, branched dendrimers, cationic liposomes and cellpenetrating peptides (CPPs). The latter represent an assortment of fairly unrelated sequences essentially characterised by a high content of basic amino acids and a length of 10-30 residues. CPPs are capable of mediating the cellular uptake of hydrophilic macromolecules like peptides and nucleic acids (e.g. siRNAs, aptamers and antisenseoligonucleotides), which are internalised by cells at a very low rate when applied alone. Up to now, numerous sequences have been reported to show cell-penetrating properties and many of them have been used to successfully transport a variety of different cargos into mammalian cells. In recent years, it has become apparent that endocytosis is a major route of internalisation even though the mechanisms underlying the cellular translocation of CPPs are poorly understood and still subject to controversial discussions. In this review, we will summarise the latest developments in peptide-based cellular delivery of nucleic acid cargos. We will discuss different mechanisms of entry, the intracellular fate of the cargo, correlation studies of uptake versus biological activity of the cargo as well as technical problems and pitfalls.
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    Fast and sensitive screening of transparent composite materials using polarized light image processing
    (Saarbrücken : Leibniz-Institut für Neue Materialien, 2009) Weiss, Ingrid M.; Heiland, Birgit
    Composite materials of natural origin have remarkable material properties. In order to mimic the various functions of such materials for technical applications, it is necessary to understand the structure and the mechanisms of their formation, a scientific field which is called structural biology. A milestone in the era of structural biology was the application of the polarized light microscope for investigating biological specimens by W. J. Schmidt in 1924. A recent development in polarization technology is the LC-PolScope (Abrio IM™ Imaging System) that has been developed by R. Oldenbourg and his colleagues during the past 20 years. Like conventional polarization microscopy, it probes the local anisotropy of the specimen’s optical properties, such as birefringence or dichroism. Since there is a close relationship between such properties and molecular order in bulk materials, polarization microscopy can be regarded as a submicroscopic technique. The LC-PolScope is especially suitable for investigating biological materials without the need for using contrasting agents such as fluorescent markers. We demonstrated the applicability of this technique for the fast and sensitive screening of biological composite materials. Implications for the characterization of biomineralization phenomena in a quantitative manner are discussed.
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    The analysis of arabidopsis nicotianamine synthase mutants reveals functions for nicotianamine in seed iron loading and iron deficiency responses
    (Rockville, MD : American Society of Plant Biologists, 2009) Fink-Straube, Claudia; Klatte, Marco; Schuler, Mara; Wirtz, Markus; Hell, Rüdiger; Bauer, Petra
    Nicotianamine chelates and transports micronutrient metal ions in plants. It has been speculated that nicotianamine is involved in seed loading with micronutrients. A tomato (Solanum lycopersicum) mutant (chloronerva) and a tobacco (Nicotiana tabacum) transgenic line have been utilized to analyze the effects of nicotianamine loss. These mutants showed early leaf chlorosis and had sterile flowers. Arabidopsis (Arabidopsis thaliana) has four NICOTIANAMINE SYNTHASE (NAS) genes. We constructed two quadruple nas mutants: one had full loss of NAS function, was sterile, and showed a chloronerva-like phenotype (nas4x-2); another mutant, with intermediate phenotype (nas4x-1), developed chlorotic leaves, which became severe upon transition from the vegetative to the reproductive phase and upon iron (Fe) deficiency. Residual nicotianamine levels were sufficient to sustain the life cycle. Therefore, the nas4x-1 mutant enabled us to study late nicotianamine functions. This mutant had no detectable nicotianamine in rosette leaves of the reproductive stage but low nicotianamine levels in vegetative rosette leaves and seeds. Fe accumulated in the rosette leaves, while less Fe was present in flowers and seeds. Leaves, roots, and flowers showed symptoms of Fe deficiency, whereas leaves also showed signs of sufficient Fe supply, as revealed by molecular-physiological analysis. The mutant was not able to fully mobilize Fe to sustain Fe supply of flowers and seeds in the normal way. Thus, nicotianamine is needed for correct supply of seeds with Fe. These results are fundamental for plant manipulation approaches to modify Fe homeostasis regulation through alterations of NAS genes.