Crystallization and preliminary X-ray analysis of sulfite dehydrogenase from Stargeya novella

Crystals of purified heterodimeric sulfite dehydrogenase from Starkeya novella have been grown using vapour diffusion. X-ray diffraction data have been collected from crystals of the native protein at lambda=1.0 Angstrom and close to the iron absorption edge at lambda=1.737 Angstrom. The crystals belong to space group P2(1)2(1)2, with unit-cell parameters a=97.5, b=92.5, c=55.9 Angstrom. Native data have been recorded to 1.8 Angstrom resolution and Fe-edge data to 2.5 Angstrom

Maturation of molybdoenzymes and its influence on the pathogenesis of non-typeable Haemophilus influenzae

© 2015 Dhouib, Pg Othman, Essilfie, Hansbro, Hanson, McEwan and Kappler. Mononuclear molybdenum enzymes of the dimethylsulfoxide (DMSO) reductase family occur exclusively in prokaryotes, and a loss of some these enzymes has been linked to a loss of bacterial virulence in several cases. The MobA protein catalyzes the final step in the synthesis of the molybdenum guanine dinucleotide (MGD) cofactor that is exclusive to enzymes of the DMSO reductase family. MobA has been proposed as a potential target for control of virulence since its inhibition would affect the activities of all molybdoenzymes dependent upon MGD. Here, we have studied the phenotype of a mobA mutant of the host-adapted human pathogen Haemophilus influenzae. H. influenzae causes and contributes to a variety of acute and chronic diseases of the respiratory tract, and several enzymes of the DMSO reductase family are conserved and highly expressed in this bacterium. The mobA mutation caused a significant decrease in the activities of all Mo-enzymes present, and also resulted in a small defect in anaerobic growth. However, we did not detect a defect in in vitro biofilm formation nor in invasion and adherence to human epithelial cells in tissue culture compared to the wild-type. In a murine in vivo model, the mobA mutant showed only a mild attenuation compared to the wild-type. In summary, our data show that MobA is essential for the activities of molybdenum enzymes, but does not appear to affect the fitness of H. influenzae. These results suggest that MobA is unlikely to be a useful target for antimicrobials, at least for the purpose of treating H. influenzae infections

Corporate Governance, Opaque Bank Activities, and Risk/Return Efficiency: Pre- and Post-Crisis Evidence from Turkey

Does better corporate governance unambiguously improve the risk/return efficiency of banks? Or does either a re-orientation of banks' revenue mix towards more opaque products, an economic downturn, or tighter supervision create off-setting or reinforcing effects? The authors relate bank efficiency to shortfalls from a stochastic risk/return frontier. They analyze how internal governance mechanisms (CEO duality, board experience, political connections, and education profile) and external governance mechanisms (discipline exerted by shareholders, depositors, or skilled employees) determine efficiency in a sample of Turkish banks. The 2000 financial crisis was a wakeup call for bank efficiency and corporate governance. As a result, better corporate governance mechanisms have been able to improve risk/return efficiency when the economic, regulatory, and supervisory environments are more stable and bank products are more complex.corporate governance;bank risk;noninterest income;crisis;frontier

The DmsABC Sulfoxide Reductase Supports Virulence in Non-typeable Haemophilus influenzae.

Although molybdenum-containing enzymes are well-established as having a key role in bacterial respiration, it is increasingly recognized that some may also support bacterial virulence. Here, we show that DmsABC, a putative dimethylsulfoxide (DMSO) reductase, is required for fitness of the respiratory pathogen Haemophilus influenzae (Hi) in different models of infection. Expression of the dmsABC operon increased with decreasing oxygen availability, but despite this, a Hi2019Δd msA strain did not show any defects in anaerobic growth on chemically defined medium (CDM), and viability was also unaffected. Although Hi2019Δd msA exhibited increased biofilm formation in vitro and greater resistance to hypochlorite killing compared to the isogenic wild-type strain, its survival in contact with primary human neutrophils, in infections of cultured tissue cells, or in a mouse model of lung infection was reduced compared to Hi2019WT. The tissue cell infection model revealed a two-fold decrease in intracellular survival, while in the mouse model of lung infection Hi2019Δd msA was strongly attenuated and below detection levels at 48 h post-inoculation. While Hi2019WT was recovered in approximately equal numbers from bronchoalveolar lavage fluid (BALF) and lung tissue, survival of Hi2019Δd msA was reduced in lung tissue compared to BALF samples, indicating that Hi2019Δd msA had reduced access to or survival in the intracellular niche. Our data clearly indicate for the first time a role for DmsABC in H. influenzae infection and that the conditions under which DmsABC is required in this bacterium are closely linked to interactions with the host

Alternative fuels from forest residues for passenger cars - an assessment under German framework conditions

Background Due to the available volumes, biogenic residues are a promising resource for renewable fuels for passenger cars to reduce greenhouse gas (GHG) emissions. In this study, we compare three fuels from forest residues under German framework conditions: biogenic electricity, substitute natural gas (SNG), and Fischer-Tropsch (FT) diesel. Methods Fuels from forest residues are compared with regard to their technical efficiency (here defined as ‘pkm per kg biomass’), costs, and environmental impacts with a focus on GHG emissions. We took into consideration the real-life driving conditions and corresponding car classes as well as market penetration scenarios for electric and gaseous fuel cars. Results Our results show that the technical efficiency of biogenic electricity is high, while the economic and environmental results strongly depend on the car size and market penetration assumptions. Furthermore, it is essential to clearly define the main goal of introducing fuels from forest residues. If the goal is to reduce GHG emissions at the lowest cost, SNG (and natural gas) in bigger cars is preferable. For high GHG reductions at the lowest forest residue consumption, biogenic electricity in smaller commuter-type cars are found to be a good solution. This also proves true for the aggregated environmental impact score ReCiPe Total. Conclusions It is important to include mobility patterns and a clear goal definition when comparing biogenic fuels. In Germany, biogenic electricity, SNG, and FT diesel can reduce GHG emissions at reduction costs of around 100 €/t CO2-Eq when used the right way

Evaluation of corrections for spherical-shell neutron transmission experiments by the Monte-Carlo technique

Berechnung von Korrekturen für Neutronen-Kugelschalentransmissions-Experimente mit der Monte-Carlo-Methode. Der Einfluß einiger experimenteller Einzelheiten auf die Meßgrößen bei Neutronen-Kugelschalentransmissionsexperimenten zur Kerndatenüberprüfung wurde abgeschätzt, indem die entsprechenden Bestandteile der Anordnung (Material nahe der Neutronenquelle, Kollimator und Abschirmung des Detektors u.a.) sowie die spezifische Meßmethode (Flugzeitspektrometrie) in Monte-Carlo-Rechnungen simuliert wurden. Die Rechnungen wurden mit dem Programm MCNP und Kerndaten der Bibliotheken EFF-1 und FENDL-1 durchgeführt. Es wurden Korrekturen bestimmt, die die Auswirkungen der verschiedener Details jeweils getrennt beschreiben

Monte Carlo Study of Cluster-Diameter Distribution: A New Observable to Estimate Correlation Lengths

We report numerical simulations of two-dimensional $q$-state Potts models with emphasis on a new quantity for the computation of spatial correlation lengths. This quantity is the cluster-diameter distribution function $G_{diam}(x)$, which measures the distribution of the diameter of stochastically defined cluster. Theoretically it is predicted to fall off exponentially for large diameter $x$, $G_{diam} \propto \exp(-x/\xi)$, where $\xi$ is the correlation length as usually defined through the large-distance behavior of two-point correlation functions. The results of our extensive Monte Carlo study in the disordered phase of the models with $q=10$, 15, and $20$ on large square lattices of size $300 \times 300$, $120 \times 120$, and $80 \times 80$, respectively, clearly confirm the theoretically predicted behavior. Moreover, using this observable we are able to verify an exact formula for the correlation length $\xi_d(\beta_t)$ in the disordered phase at the first-order transition point $\beta_t$ with an accuracy of about $1$ for all considered values of $q$. This is a considerable improvement over estimates derived from the large-distance behavior of standard (projected) two-point correlation functions, which are also discussed for comparison.Comment: 20 pages, LaTeX + 13 postscript figures. See also http://www.cond-mat.physik.uni-mainz.de/~janke/doc/home_janke.htm

SoxAX cytochromes, a new type of heme copper protein involved in bacterial energy generation from sulfur compounds

SoxAX cytochromes are essential for the function of the only confirmed pathway for bacterial thiosulfate oxidation, the so-called "Sox pathway," in which they catalyze the initial formation of a S-S bond between thiosulfate and the SoxYZ carrier protein. Our work using the Starkeya novella diheme SoxAX protein reveals for the first time that in addition to two active site heme groups, SoxAX contains a mononuclear Cu(II) center with a distorted tetragonal geometry and three to four nitrogen ligands, one of which is a histidine. The Cu(II) center enhanced SoxAX activity in a newly developed, glutathione-based assay system that mimics the natural reaction of SoxAX with SoxYZ. EPR spectroscopy confirmed that the SoxAX CuII center is reduced by glutathione. At pH 7 a K(m app) of 0.19 +/- 0.028 mM and a k(cat app) of 5.7 +/- 0.25s(-1) were determined for glutathione. We propose that SoxAX cytochromes are a new type of heme-copper proteins, with SoxAX-mediated S-S bond formation involving both the copper and heme centers

A weak characterization of slow variables in stochastic dynamical systems

We present a novel characterization of slow variables for continuous Markov processes that provably preserve the slow timescales. These slow variables are known as reaction coordinates in molecular dynamical applications, where they play a key role in system analysis and coarse graining. The defining characteristics of these slow variables is that they parametrize a so-called transition manifold, a low-dimensional manifold in a certain density function space that emerges with progressive equilibration of the system's fast variables. The existence of said manifold was previously predicted for certain classes of metastable and slow-fast systems. However, in the original work, the existence of the manifold hinges on the pointwise convergence of the system's transition density functions towards it. We show in this work that a convergence in average with respect to the system's stationary measure is sufficient to yield reaction coordinates with the same key qualities. This allows one to accurately predict the timescale preservation in systems where the old theory is not applicable or would give overly pessimistic results. Moreover, the new characterization is still constructive, in that it allows for the algorithmic identification of a good slow variable. The improved characterization, the error prediction and the variable construction are demonstrated by a small metastable system

Influenza nucleoprotein delivered with aluminium salts protects mice from an influenza virus that expresses an altered nucleoprotein sequence

Influenza virus poses a difficult challenge for protective immunity. This virus is adept at altering its surface proteins, the proteins that are the targets of neutralizing antibody. Consequently, each year a new vaccine must be developed to combat the current recirculating strains. A universal influenza vaccine that primes specific memory cells that recognise conserved parts of the virus could prove to be effective against both annual influenza variants and newly emergent potentially pandemic strains. Such a vaccine will have to contain a safe and effective adjuvant that can be used in individuals of all ages. We examine protection from viral challenge in mice vaccinated with the nucleoprotein from the PR8 strain of influenza A, a protein that is highly conserved across viral subtypes. Vaccination with nucleoprotein delivered with a universally used and safe adjuvant, composed of insoluble aluminium salts, provides protection against viruses that either express the same or an altered version of nucleoprotein. This protection correlated with the presence of nucleoprotein specific CD8 T cells in the lungs of infected animals at early time points after infection. In contrast, immunization with NP delivered with alum and the detoxified LPS adjuvant, monophosphoryl lipid A, provided some protection to the homologous viral strain but no protection against infection by influenza expressing a variant nucleoprotein. Together, these data point towards a vaccine solution for all influenza A subtypes