Q fever: baseline monitoring of a sheep and a goat flock associated with human infections.

Animal losses due to abortion and weak offspring during a lambing period amounted up to 25% in a goat flock and up to 18% in a sheep flock kept at an experimental station on the Swabian Alb, Germany. Fifteen out of 23 employees and residents on the farm tested positive for Coxiella burnetii antibodies by enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence assay. Ninety-four per cent of the goats and 47% of the sheep were seropositive for C. burnetii by ELISA. Blood samples of 8% of goats and 3% of sheep were PCR positive. C. burnetii was shed by all tested animals through vaginal mucus, by 97% of the goats and 78% of the sheep through milk, and by all investigated sheep through faeces (PCR testing). In this outbreak human and animal infection were temporally related suggesting that one was caused by the other.

Semimetal to semiconductor transition in Bi/TiO2 core/shell nanowires.

We demonstrate the full thermoelectric and structural characterization of individual bismuth-based (Bi-based) core/shell nanowires. The influence of strain on the temperature dependence of the electrical conductivity, the absolute Seebeck coefficient and the thermal conductivity of bismuth/titanium dioxide (Bi/TiO2) nanowires with different diameters is investigated and compared to bismuth (Bi) and bismuth/tellurium (Bi/Te) nanowires and bismuth bulk. Scattering at surfaces, crystal defects and interfaces between the core and the shell reduces the electrical conductivity to less than 5% and the thermal conductivity to less than 25% to 50% of the bulk value at room temperature. On behalf of a compressive strain, Bi/TiO2 core/shell nanowires show a decreasing electrical conductivity with decreasing temperature opposed to that of Bi and Bi/Te nanowires. We find that the compressive strain induced by the TiO2 shell can lead to a band opening of bismuth increasing the absolute Seebeck coefficient by 10% to 30% compared to bulk at room temperature. In the semiconducting state, the activation energy is determined to |41.3 ± 0.2| meV. We show that if the strain exceeds the elastic limit the semimetallic state is recovered due to the lattice relaxation.

Vaccinia virus anti-apoptotic F1L is a novel Bcl-2-like domain-swapped dimer that binds a highly selective subset of BH3-containing death ligands.

Apoptosis is an important part of the host's defense mechanism for eliminating invading pathogens. Some viruses express proteins homologous in sequence and function to mammalian pro-survival Bcl-2 proteins. Anti-apoptotic F1L expressed by vaccinia virus is essential for survival of infected cells, but it bears no discernable sequence homology to proteins other than its immediate orthologues in related pox viruses. Here we report that the crystal structure of F1L reveals a Bcl-2-like fold with an unusual N-terminal extension. The protein forms a novel domain-swapped dimer in which the alpha1 helix is the exchanged domain. Binding studies reveal an atypical BH3-binding profile, with sub-micromolar affinity only for the BH3 peptide of pro-apoptotic Bim and low micromolar affinity for the BH3 peptides of Bak and Bax. This binding interaction is sensitive to F1L mutations within the predicted canonical BH3-binding groove, suggesting parallels between how vaccinia virus F1L and myxoma virus M11L bind BH3 domains. Structural comparison of F1L with other Bcl-2 family members reveals a novel sequence signature that redefines the BH4 domain as a structural motif present in both pro- and anti-apoptotic Bcl-2 members, including viral Bcl-2-like proteins.

Nanometrology: Absolute Seebeck coefficient of individual silver nanowires.

Thermoelectric phenomena can be strongly modified in nanomaterials. The determination of the absolute Seebeck coefficient is a major challenge for metrology with respect to micro- and nanostructures due to the fact that the transport properties of the bulk material are no more valid. Here, we demonstrate a method to determine the absolute Seebeck coefficient S of individual metallic nanowires. For highly pure and single crystalline silver nanowires, we show the influence of nanopatterning on S in the temperature range between 16 K and 300 K. At room temperature, a nanowire diameter below 200 nm suppresses S by 50% compared to the bulk material to less than S = 1 µVK-1, which is attributed to the reduced electron mean free path. The temperature dependence of the absolute Seebeck coefficient depends on size effects. Thermodiffusion and phonon drag are reduced with respect to the bulk material and the ratio of electron-phonon to phonon-phonon interaction is significantly increased.

Frequent loss of expression of the pro-apoptotic protein Bim in renal cell carcinoma: evidence for contribution to apoptosis resistance.

Renal cell carcinoma (RCC) is resistant to chemotherapy, and this resistance is mirrored by a high apoptosis resistance of many RCC lines in vitro. Here, we report the loss of the pro-apoptotic BH3-only protein Bim in a large part of clinical RCC cases and provide evidence for a functional relevance of this loss. Immunohistochemistry of clear cell renal cell carcinoma cases and corresponding normal kidney showed strong Bim reactivity in renal tubules of all cases but loss of Bim in 35 of 45 RCC samples. Out of nine RCC cell lines investigated, six showed strongly diminished or undetectable levels of Bim protein by western blotting. Four RCC lines of varying apoptosis sensitivity were analysed further. Bcl-2, Bcl-x(L), Mcl-1, Bax and Bak expression did not correlate with apoptosis sensitivity. All cell lines underwent apoptosis upon forced expression of Bax and Bim, suggesting an upstream difference. In all four lines, adriamycin induced p53 but not its targets Puma or Noxa. However, apoptosis sensitivity correlated with levels of Bim protein. Bim siRNA reduced apoptosis sensitivity in a susceptible cell line. Furthermore, inhibition of histone deacetylation restored Bim expression in cell lines. These data suggest that Bim has a function as a tumor suppressor in RCC.

Modified vaccinia virus Ankara protein F1L is a novel BH3-domain-binding protein and acts together with the early viral protein E3L to block virus-associated apoptosis.

Infection with viruses often protects the infected cell against external stimuli to apoptosis. Here we explore the balance of apoptosis induction and inhibition for infection with the modified vaccinia virus Ankara (MVA), using two MVA mutants with experimentally introduced deletions. Deletion of the E3L-gene from MVA transformed the virus from an inhibitor to an inducer of apoptosis. Noxa-deficient mouse embryonic fibroblasts (MEF) were resistant to MVA-DeltaE3L-induced apoptosis. When the gene encoding F1L was deleted from MVA, apoptosis resulted that required Bak or Bax. MVA-DeltaF1L-induced apoptosis was blocked by Bcl-2. When expressed in HeLa cells, F1L blocked apoptosis induced by forced expression of the BH3-only proteins, Bim, Puma and Noxa. Finally, biosensor analysis confirmed direct binding of F1L to BH3 domains. These data describe a molecular framework of how a cell responds to MVA infection by undergoing apoptosis, and how the virus blocks apoptosis by interfering with critical steps of its signal transduction.

Biomimetic cellular metals-using hierarchical structuring for energy absorption.

Fruit walls as well as nut and seed shells typically perform a multitude of functions. One of the biologically most important functions consists in the direct or indirect protection of the seeds from mechanical damage or other negative environmental influences. This qualifies such biological structures as role models for the development of new materials and components that protect commodities and/or persons from damage caused for example by impacts due to rough handling or crashes. We were able to show how the mechanical properties of metal foam based components can be improved by altering their structure on various hierarchical levels inspired by features and principles important for the impact and/or puncture resistance of the biological role models, rather than by tuning the properties of the bulk material. For this various investigation methods have been established which combine mechanical testing with different imaging methods, as well as with in situ and ex situ mechanical testing methods. Different structural hierarchies especially important for the mechanical deformation and failure behaviour of the biological role models, pomelo fruit (Citrus maxima) and Macadamia integrifolia, were identified. They were abstracted and transferred into corresponding structural principles and thus hierarchically structured bio-inspired metal foams have been designed. A production route for metal based bio-inspired structures by investment casting was successfully established. This allows the production of complex and reliable structures, by implementing and combining different hierarchical structural elements found in the biological concept generators, such as strut design and integration of fibres, as well as by minimising casting defects. To evaluate the structural effects, similar investigation methods and mechanical tests were applied to both the biological role models and the metallic foams. As a result an even deeper quantitative understanding of the form-structure-function relationship of the biological concept generators as well as the bio-inspired metal foams was achieved, on deeper hierarchical levels and overarching different levels.

Characterization of antiapoptotic activities of Chlamydia pneumoniae in infected cells.

C. pneumoniae has the capacity to inhibit apoptosis. Here we provide further insight into how infection with C. pneumoniae affects the apoptotic signal transduction in infected human epitheloid cells. For assessing changes in the apoptotic response, intact cells and cell extracts were used. These results show that C. pneumoniae is able to interfere with the host cell's apoptotic apparatus at least at two steps in the signal transduction.

Theory of electron localization at dielectric-metal interfaces: a continuum model.

Localization of electrons at dielectric-metal interfaces is studied in the framework of a continuum model. The layer of thickness L, with a negative electron affinity, is characterized by the static dielectric constant epsilons and by the optical dielectric constant epsiloninfinity. It is found that the electron localization along the plane of the interface occurs if the layer thickness exceeds a critical value Lc. In the case of a high polar layer, the electron energy of the localized ground state shows a nonmonotonic dependence on the layer thickness. A strong correlation between low-lying excitations and the spread of the localized state has been established. The magnitude of the correlation parameter is close to the analogous correlation for the solvated electron in the bulk. The localization dynamics is discussed in terms of relaxation along a polarization coordinate, which is directly connected to the polarization energy of the layer.

Characterization of antiapoptotic activities of Chlamydia pneumoniae in human cells.

Chlamydia pneumoniae is an obligate intracellular bacterium which frequently causes airway infection in humans and has been implicated in atherosclerosis. Here we show that infection with C. pneumoniae protects HeLa human epithelioid cells against apoptosis induced by external stimuli. In infected HeLa cells, apoptosis induced by staurosporine and CD95-death-receptor signaling was strongly reduced. Upon treatment with staurosporine, generation of effector caspase activity, processing of caspase-3 and caspase-9 and cytochrome c redistribution were all profoundly inhibited in cells infected with C. pneumoniae. Bacterial protein synthesis during early infection was required for this inhibition. Furthermore, cytochrome c-induced processing and activation of caspases were inhibited in cytosolic extracts from infected cells, suggesting that a C. pneumoniae-dependent antiapoptotic factor was generated in the cytosol upon infection. Infection with C. pneumoniae failed to induce significant NF-kappaB activation in HeLa cells, indicating that no NF-kappaB-dependent cellular factors were involved in the protection against apoptosis. These results show that C. pneumoniae is capable of interfering with the host cell's apoptotic apparatus at probably at least two steps in signal transduction and might explain the propensity of these bacteria to cause chronic infections in humans.

Trienoic fatty acids are required to maintain chloroplast function at low temperatures.

The chloroplast membranes of all higher plants contain very high proportions of trienoic fatty acids. To investigate how these lipid structures are important in photosynthesis, we have generated a triple mutant line of Arabidopsis that contains negligible levels of trienoic fatty acids. For mutant plants grown at 22 degrees C, photosynthetic fluorescence parameters were indistinguishable from wild type at 25 degrees C. Lowering the measurement temperature led to a small decrease in photosynthetic quantum yield, Phi(II), in the mutant relative to wild-type controls. These and other results indicate that low temperature has only a small effect on photosynthesis in the short term. However, long-term growth of plants at 4 degrees C resulted in decreases in fluorescence parameters, chlorophyll content, and thylakoid membrane content in triple-mutant plants relative to wild type. Comparisons among different mutant lines indicated that these detrimental effects of growth at 4 degrees C are strongly correlated with trienoic fatty acid content with levels of 16:3 + 18:3, approximately one-third of wild type being sufficient to sustain normal photosynthetic function. In total, our results indicate that trienoic fatty acids are important to ensure the correct biogenesis and maintenance of chloroplasts during growth of plants at low temperatures.

Molecular basis for pH sensitivity and proton transfer in green fluorescent protein: protonation and conformational substates from electrostatic calculations.

We performed a theoretical study to elucidate the coupling between protonation states and orientation of protein dipoles and buried water molecules in green fluorescent protein, a versatile biosensor for protein targeting. It is shown that the ionization equilibria of the wild-type green fluorescent protein-fluorophore and the internal proton-binding site E222 are mutually interdependent. Two acid-base transitions of the fluorophore occur in the presence of neutral (physiologic pH) and ionized (pH > 12) E222, respectively. In the pH-range from approximately 8 to approximately 11 ionized and neutral sites are present in constant ratio, linked by internal proton transfer. The results indicate that modulation of the internal proton sharing by structural fluctuations or chemical variations of aligning residues T203 and S65 cause drastic changes of the neutral/anionic ratio-despite similar physiologic fluorophore pK(a) s. Moreover, we find that dipolar heterogeneities in the internal hydrogen-bond network lead to distributed driving forces for excited-state proton transfer. A molecular model for the unrelaxed surrounding after deprotonation is discussed in relation to pathways providing fast ground-state recovery or slow stabilization of the anion. The calculated total free energy for excited-state deprotonation ( approximately 19 k(B)T) and ground-state reprotonation ( approximately 2 k(B)T) is in accordance with absorption and emission data (

Analysis of optical spectra from single crystals of Rhodopseudomonas viridis reaction centers.

Absorption spectra and light-induced absorbance changes of crystals from Rhodopseudomonas viridis reaction centers are recorded. A theoretical analysis of the absorption and circular dichroism spectra is presented, yielding a consistent picture of spectroscopic and structural information.