Scientific publications

BACKGROUND: Mollusc shells are commonly investigated using high-resolution imaging techniques based on cryo-fixation. Less detailed information is available regarding the light-optical properties. Sea shells of Haliotis pulcherina were embedded for polishing in defined orientations in order to investigate the interface between prismatic calcite and nacreous aragonite by standard materialographic methods. A polished thin section of the interface was prepared with a defined thickness of 60 µm for quantitative birefringence analysis using polarized light and LC-PolScope microscopy. Scanning electron microscopy images were obtained for comparison. In order to study structural-mechanical relationships, nanoindentation experiments were performed.RESULTS:Incident light microscopy revealed a super-structure in semi-transparent regions of the polished cross-section under a defined angle. This super-structure is not visible in transmitted birefringence analysis due to the blurred polarization of small nacre platelets and numerous organic interfaces. The relative orientation and homogeneity of calcite prisms was directly identified, some of them with their optical axes exactly normal to the imaging plane. Co-oriented "prism colonies" were identified by polarized light analyses. The nacreous super-structure was also visualized by secondary electron imaging under defined angles. The domains of the super-structure were interpreted to consist of crystallographically aligned platelet stacks. Nanoindentation experiments showed that mechanical properties changed with the same periodicity as the domain size.CONCLUSIONS:In this study, we have demonstrated that insights into the growth mechanisms of nacre can be obtained by conventional light-optical methods. For example, we observed super-structures formed by co-oriented nacre platelets as previously identified using X-ray Photo-electron Emission Microscopy (X-PEEM) [Gilbert et al., Journal of the American Chemical Society 2008, 130:17519–17527]. Polarized optical microscopy revealed unprecedented super-structures in the calcitic shell part. This bears, in principle, the potential for in vivo studies, which might be useful for investigating the growth modes of nacre and other shell types.

We analyze theoretically the moment of inertia of the desert ant Cataglyphis (C. bicolor and C. fortis) around a vertical axis through its own center of mass when the animal raises its gaster to a vertical position. Compared to the value when the gaster is horizontal, the moment of inertia is reduced to one half; this implies that when increasing its angular acceleration the ant need apply only half the level of torque when the gaster is raised, compared to when the gaster is lowered. As an example, we analyze the cases of an ant running on circular and sinusoidal paths. In both cases, the ant must apply a sideways thrust, anti-roll and anti-pitch torques to avoid toppling, and, on the circular path when accelerating and throughout the sinusoidal trajectory, a torque to enable turning as the path curves. When the ant is accelerating in a very tight circle or running on a very narrow sinusoidal path, in which the amplitude of the sinusoid is less than the length of the ant's body, the forces required for the turning torque can equal and exceed those required for the sideways thrust, and can be reduced significantly by the ant raising the gaster, whereas the foot-thrust for the anti-roll and anti-pitch torques rises only modestly when the gaster is up. This suggests that there may be an evolutionary advantage for employing the gaster-raising mode of locomotion, since this habit will allow desert ants to use lower forces and less energy, and perhaps run faster on more tortuous paths.

Localization and intracellular migration of 32 and 83 nm SiO2 nanoparticles in relation to the nucleus was evaluated in vitro on undifferentiated human colon carcinoma (Caco-2) cells. The fluorescence dye Atto647N was incorporated into the particles, which enabled detection by high resolution, nondiffraction limited stimulated emission depletion (STED) microscopy. The distribution and agglomeration of nanoparticles was measured with STED microscopy after 5, 24, 48, and 72 h. Analyses revealed that only 32 nm silica particles penetrated into the nucleus of Caco-2 cells upon exposure for 24 h. Here, they formed agglomerates up to 300 nm after 72 h of incubation. Quantitative evaluation of the migration of 32 nm compared to 83 nm particles demonstrated that 32 nm particles obviously migrated faster into and through the cell in the beginning (5 h time point). The presence and agglomeration inside the cells and the penetration into the nucleus were considered to potentially activate cytotoxic responses. Therefore, the cytotoxic (WST-1 assay) and genotoxic (comet assay) effects of the silica nanoparticles were evaluated. Even though 32 nm silica particles are penetrating into the nucleus, neither cytotoxic nor genotoxic effects were detected for either particle size.

We describe here a chemically controlled pathway for the designed synthesis of iron oxide nanoparticles by thermal decomposition of iron(II) and iron(III) oxalates in high-boiling solvents in the presence of oleylamine and oleic acid acting as capping ligands. The phase composition of the nanocrystals (Fe, FeO, Fe3O4, or α-Fe2O3) could be precisely controlled by adjusting the synthesis conditions or by addition of appropriate oxidants, such as trimethylamine-N-oxide (TMAO), which produced highly monodisperse iron(III) oxide particles in the range of 6-25 nm in good yields. The decomposition behavior of different precursor/TMAO mixtures was elucidated by differential scanning calorimetry and thermogravimetry, and resulting particles were characterized by comprehensive HR-TEM and XRD analyses.

Colloidal particles are continuously assembled into crystalline particle coatings using convective fluid flows. Assembly takes place inside a meniscus on a wetting reservoir. The shape of the meniscus defines the profile of the convective flow and the motion of the particles. We use optical interference microscopy, particle image velocimetry and particle tracking to analyze the particles? trajectory from the liquid reservoir to the film growth front and inside the deposited film as a function of temperature. Our results indicate a transition from assembly at a static film growth front at high deposition temperatures to assembly in a precursor film with high particle mobility at low deposition temperatures. A simple model that compares the convective drag on the particles to the thermal agitation explains this behavior. Convective assembly mechanisms exhibit a pronounced temperature dependency and require a temperature that provides sufficient evaporation. Capillary mechanisms are nearly temperature independent and govern assembly at lower temperatures. The model fits the experimental data with temperature and particle size as variable parameters and allows prediction of the transition temperatures. While the two mechanisms are markedly different, dried particle films from both assembly regimes exhibit hexagonal particle packings. We show that films assembled by convective mechanisms exhibit greater regularity than those assembled by capillary mechanisms.

Monodisperse, crystalline silver nanoparticles are synthesized by sonolytically assisted decomposition of silver oxalate and oleate precursors in dibenzylether. Oleic acid and oleylamine are used as capping agents. The diameters of the obtained particles vary from 6 to 10 nm depending on the sonification time and the precursor material. The particles are stable in non-polar media. A 2D and 3D assembly of the as-prepared nanoparticles is observed. Particle sizes and dispersities are characterized by transmission electron microscopy and image analysis. The decomposition characteristics of the different precursor compounds are investigated using UV–Vis spectroscopy and thermogravimetry.

The metal chelator nicotianamine promotes the bioavailability of Fe and reduces cellular Fe toxicity. For breeding Fe-efficient crops, we need to explore the fundamental impact of nicotianamine on plant development and physiology. The quadruple nas4x-2 mutant of Arabidopsis thaliana cannot synthesize any nicotianamine, shows strong leaf chlorosis, and is sterile. To date, these phenotypes have not been fully explained. Here, we show that sink organs of this mutant were Fe deficient, while aged leaves were Fe sufficient. Upper organs were also Zn deficient. We demonstrate that transport of Fe to aged leaves relied on citrate, which partially complemented the loss of nicotianamine. In the absence of nicotianamine, Fe accumulated in the phloem. Our results show that rather than enabling the long-distance movement of Fe in the phloem (as is the case for Zn), nicotianamine facilitates the transport of Fe from the phloem to sink organs. We delimit nicotianamine function in plant reproductive biology and demonstrate that nicotianamine acts in pollen development in anthers and pollen tube passage in the carpels. Since Fe and Zn both enhance pollen germination, a lack of either metal may contribute to the reproductive defect. Our study sheds light on the physiological functions of nicotianamine.

In biological cells, various transmembrane enzymes function as highly effective chemical reactors confined in space with characteristic length scales of tens of nanometers to micrometer. However, it is still challenging to quantitatively confine membranes in compact reactor platforms without losing their biochemical functions. Here, a simple and straightforward strategy towards the fabrication of a new flow-through reactor by the functional coating of porous silica microparticles with sarcoplasmic reticulum membranes is described. After a short incubation, the membranes achieve the homogeneous, full coverage of the particle surface, spanning across pores with the diameter of about 100 nm. By using the underlying pores as cavity reservoirs, transmembrane enzyme (Ca2+-ATPase) in the membrane retains their capability of ATP hydrolysis. This enables us to confine 1.1 m2 of native membranes containing a large amount of Ca2+-ATPase (approx. 10 nmol) in a column-packaged, flow-through reactor with merely 1.8 mL volume, which cannot be achieved by the reconstitution of proteins in artificial lipid membranes or condensation of membranes in suspensions. The distinct functional levels corresponding to different reaction buffers can be reproduced even after many buffer exchanges over 14 days, confirming the stability and reproducibility of the membrane-particle hybrid reactors.

The synthesis, structural characterization, electrochemistry and luminescence properties of a series of new yttrium and europium(+3) alkoxides bearing thiophene moieties are presented. The yttrium compounds were obtained by the reaction between Y[N(SiMe3)2]3 and the tertiary alcohols HO–C(C16H13S) (2), HO–C(C17H15S) (3), HO–C(C14H11S2) (4) and HO–C(C16H13S) (5) in thf or in a mixture of toluene and pyridine. The X-ray crystal diffraction measurements show a five-coordinated yttrium atom in distorted trigonal-bipyramidal geometry. The metal centres are surrounded by threemethoxido ligands in equatorial positions and two tetrahydrofuran [for {Y[OC(C16H13S)]3(thf)2}·toluene (8), {Y[OC(C17H15S)]3(thf)2}·toluene (10) and {Y[OC(C14H11S2)]3(thf)2}·1/2 toluene (12)] or two pyridine [for {Y[OC(C16H13S)]3(py)2}·toluene (9) and {Y[OC(C17H15S)]3(py)2}·toluene (11)] molecules in axial positions. The compounds Y[OC(C14H11S2)]3(py)2 (13) and Y[OC(C16H13S)]3(py)2 (14) were identified by NMR spectroscopy. In addition, a novel europium(+3) alkoxide {Eu[OC(C4H3S)3]3(thf)3}·thf (15) was synthesized by the reaction between Eu[N(SiMe3)2]3 and the tertiary alcohol HO–C(C4H3S)3 (1) in thf. The molecular structure of this compound reveals an approximately octahedral coordination sphere around the europium(+3) metal centre with three methoxido ligands and three facially arranged tetrahydrofuran molecules. The cyclic voltammograms of the yttrium alkoxides indicate that the electrochemical properties are essentially dominated by the organic ligands. The electrochemical properties of {Eu[OC(C4H3S)3]3(thf)3}·thf (15) are dominated by the oxidation of the thienyl moieties and in addition a reduction wave due to the reduction of Eu3+ to Eu2+ is visible. In comparison to the carbinols, the oxidation peak potentials of the thienyl units for the yttrium and europium(3+) alkoxides are marginally shifted towards higher values. The emission spectra of the carbinols and their derived yttrium compounds display broad bands attributed to the π*→π transitions of the aromatic ligands. Luminescence studies performed on compound 15 reveal the typical f–f transitions of the Eu3+ ions and suggest that an energy transfer from the ligand to the metal atom operates.

Insolubility is one of the possible functions of proteins involved in biomineralization, which often limits their native purification. This becomes a major problem especially when recombinant expression systems are required to obtain larger amounts. For example, the mollusc shell provides a rich source of unconventional proteins, which can interfere in manifold ways with different mineral phases and interfaces. Therefore, the relevance of such proteins for biotechnological processes is still in its infancy. Here we report a simple and reproducible purification procedure for a GFP-tagged lectin involved in biomineralization, originally isolated from mother-of-pearl in abalone shells. An optimization of E. coli host cell culture conditions was the key to obtain reasonable yields and high degrees of purity by using simple one-step affinity chromatography. We identified a dual functional role for the GFP domain when it became part of a mineralizing system in vitro. First, the GFP domain improved the solubility of an otherwise insoluble protein, in this case recombinant perlucin derivatives. Second, GFP inhibited calcium carbonate precipitation in a concentration dependent manner. This was demonstrated here using a simple bulk assay over a time period of 400 seconds. At concentrations of 2 mg/ml and higher, the inhibitory effect was observed predominantly for HCO3−as the first ionic interaction partner, but not necessarily for
Ca2+. The interference of GFP-tagged perlucin derivatives with the precipitation of calcium carbonate generated different types of GFP-fluorescent composite calcite crystals. GFP-tagging offers therefore a genetically tunable tool to gently modify mechanical and optical properties of synthetic biocomposite minerals.