Unconventional specimen preparation techniques using high resolution low voltage field emission scanning electron microscopy to study cell motility, host cell invasion, and internal cell structures in Toxoplasma gondii.

Apicomplexan parasites employ complex and unconventional mechanisms for cell locomotion, host cell invasion, and cell division that are only poorly understood. While immunofluorescence and conventional transmission electron microscopy have been used to answer questions about the localization of some cytoskeletal proteins and cell organelles, many questions remain unanswered, partly because new methods are needed to study the complex interactions of cytoskeletal proteins and organelles that play a role in cell locomotion, host cell invasion, and cell division. The choice of fixation and preparation methods has proven critical for the analysis of cytoskeletal proteins because of the rapid turnover of actin filaments and the dense spatial organization of the cytoskeleton and its association with the complex membrane system. Here we introduce new methods to study structural aspects of cytoskeletal motility, host cell invasion, and cell division of Toxoplasma gondii, a most suitable laboratory model that is representative of apicomplexan parasites. The novel approach in our experiments is the use of high resolution low voltage field emission scanning electron microscopy (LVFESEM) combined with two new specimen preparation techniques. The first method uses LVFESEM after membrane extraction and stabilization of the cytoskeleton. This method allows viewing of actin filaments which had not been possible with any other method available so far. The second approach of imaging the parasite's ultrastructure and interactions with host cells uses semithick sections (200 nm) that are resin de-embedded (Ris and Malecki, 1993) and imaged with LVFESEM. This method allows analysis of structural detail in the parasite before and after host cell invasion and interactions with the membrane of the parasitophorous vacuole as well as parasite cell division.     

Controlled self-assembly of re-engineered insulin by Fe(II)

Self-assembly of proteins mediated by metal ions is crucial in biological systems and a better understanding and novel strategies for its control are important. An abiotic metal ion ligand in a protein offers the prospect of control of the oligomeric state, if a selectivity over binding to the native side chains can be achieved. Insulin binds Zn(II) to form a hexamer, which is important for its storage in vivo and in drug formulations. We have re-engineered an insulin variant to control its self-assembly by covalent attachment of 2,2'-bipyridine. The use of Fe(II) provided chemoselective binding over the native site, forming a homotrimer in a reversible manner, which was easily followed by the characteristic color of the Fe(II) complex. This provided the first well-defined insulin trimer and the first insulin variant for which self-assembly can be followed visually.     

Nachweis und Bestimmung der Benzo?s?ure in Lebensmitteln

Ohne Zusammenfassung     

The thermodynamic limit for jellium

The thermodynamic limit of the free energy, energy, pressure, and entropy is established for a neutral system of charged particles interacting with a fixed, uniformly charged background (jellium).     

Clustering for algebraicK-systems

We prove that for a von Neumann algebra that is an algebraicK system with respect to some automorphism, the invariant state isK-clustering andr-clustering. Further, we study by using examples how far the von Neumann algebra inherits theK property from the underlyingC ^* algebra.     

Zur Thermischen Zersetzung von CaOP3F · 2 H2O

On the Thermal Decomposition of CaPO₃F · 2H₂O . The thermal decomposition of CaPO₃F · 2H₂O was investigated by thermogravimetry under inert conditions. A parallel mass spectrometric analysis of gases produced is made. Using an effusion cell a quasi equilibrium evaporation in the nearness of the ion source of the spectrometer is achieved. The results are comparable with the thermogravimetric analysis under normal pressure. During first state of thermal decomposition one mole H₂O is lost. Then the further course is determined by release of HF and POF₃. The several steps of decomposition leading to α-Ca₂P₂O₇ at temperatures above 360° C are discussed.