Staphylococcus aureus Exfoliative Toxin E,Oligomeric State and Flip of P186: Implications for Its Action Mechanism

Staphylococcal exfoliative toxins (ETs) are glutamyl endopeptidases that specifically cleave the Glu381-Gly382 bond in the ectodomains of desmoglein 1 (Dsg1) via complex action mechanisms. To date, four ETs have been identified in different Staphylococcus aureus strains and ETE is the most recently characterized. The unusual properties of ETs have been attributed to a unique structural feature, i.e., the 180° flip of the carbonyl oxygen (O) of the nonconserved residue 192/186 (ETA/ETE numbering), not conducive to the oxyanion hole formation. We report the crystal structure of ETE determined at 1.61 Å resolution, in which P186(O) adopts two conformations displaying a 180° rotation. This finding, together with free energy calculations, supports the existence of a dynamic transition between the conformations under the tested conditions. Moreover, enzymatic assays showed no significant differences in the esterolytic efficiency of ETE and ETE/P186G, a mutant predicted to possess a functional oxyanion hole, thus downplaying the influence of the flip on the activity. Finally, we observed the formation of ETE homodimers in solution and the predicted homodimeric structure revealed the participation of a characteristic nonconserved loop in the interface and the partial occlusion of the protein active site, suggesting that monomerization is required for enzymatic activity.     

Contribution of a disulfide bridge to the stability of 1,3-1,4-P-D-glucan 4-glucanohydrolase from Bacillus licheniformis

Bacillus 1,3-1,4-ß-glucanases possess a highly conserveddisulfide bridge connecting a ß-strand with a solventexposedloop lying on top of the extended binding site cleft The contributionof the disulfide bond and of both individual cysteines (Cys61and Cys90) in the Bacillus licheniformis enzyme to stabilityand activity has been evaluated by protein engineering methods.Reduction of the disulfide bond has no effect on kinetic parameters,has only a minor effect on the activity-temperature profileat high temperatures, and destabilizes the protein by less than0.7 kcal/mol as measured by equilibrium urea denatu ration at37°C. Replacing either of the Cys residues with Ala destabilizesthe protein and lowers the specific activity. C90A retains 70%of wild-type (wt) activity (in terms of Vmax), whereas C61Aand the double mutant C61A–C90A have 10% of wt V_max. Alarger change in free energy of unfolding is seen by equilibriumurea denaturation for the C61A mutation (loop residue, 3.2 kcal/molrelative to reduced wt) as compared with the C90A mutation (ß-strandresidue, 1.8 kcal/mol relative to reduced wt), while the doublemutant C61A–C90A is 0.8 kcal/mol less stable than thesingle C61A mutant. The effects on stability are interpretedas a result of the change in hydrophobic packing that occursupon removal of the sulfur atoms in the Cys to Ala mutations     

Influence of Particle Size on the Drying Kinetics of Single Droplets Containing Mixtures of Nanoparticles and Microparticles: Modeling and Pilot-Scale Validation

The particle size of the primary particles is an important parameter influencing the drying behavior of droplets. In this work, the influence of particle size on the drying kinetics and grain properties was analyzed for droplets containing silica nanoparticles, microparticles, and mixtures of the two. The presence of microparticles was found to increase the drying rate and shrinkage of the droplet. The drying curves were modeled using a reaction engineering approach (REA) model. Finally, different suspensions were dried in a pilot-scale spray dryer in order to prove the influence of the particle size obtained in the levitator tests.     

Neuroprotective Effect of Melatonin: A Novel Therapy against Perinatal Hypoxia-Ischemia

One of the most common causes of mortality and morbidity in children is perinatal hypoxia-ischemia (HI). In spite of the advances in neonatology, its incidence is not diminishing, generating a pediatric population that will require an extended amount of chronic care throughout their lifetime. For this reason, new and more effective neuroprotective strategies are urgently required, in order to minimize as much as possible the neurological consequences of this encephalopathy. In this sense, interest has grown in the neuroprotective possibilities of melatonin, as this hormone may help to maintain cell survival through the modulation of a wide range of physiological functions. Although some of the mechanisms by which melatonin is neuroprotective after neonatal asphyxia remain a subject of investigation, this review tries to summarize some of the most recent advances related with its use as a therapeutic drug against perinatal hypoxic-ischemic brain injury, supporting the high interest in this indoleamine as a future feasible strategy for cerebral asphyctic events.     

Ionic liquid recovery alternatives in ionic liquid‐based three‐phase partitioning (ILTPP)

Ionic liquid‐based three‐phase partitioning (ILTPP) is a promising technique to recover high‐added value proteins at the liquid–liquid interface. Its economic and environmental performance highly depends on the net ionic liquid consumption. Alternatives to maximize the fraction of ionic liquid that can be recycled are studied. It is demonstrated that the addition of extra salt, previously proposed in literature, has a very limited effect on ionic liquid recovery for relatively high protein concentrations in the feed stream, and that it may even lead to an increase of the ionic liquid losses under certain conditions. However, small additions of salt are shown to be effective and profitable from an economic point of view. Vacuum evaporation is shown to allow for the complete ionic liquid and salt recovery, reinforcing the sustainability and viability of ILTPP processes. © 2014 American Institute of Chemical Engineers AIChE J, 60: 3577–3586, 2014     

Effect of Zn addition on the structure and electrochemical properties of co-doped BaCe0.6Zr0.2Ln0.2O3-δ (Ln=Y,Gd, Yb) proton conductors

In this work, BaCe_0.6Zr_0.2Y_0.2-xYb_xO_3-δ and BaCe_0.6Zr_0.2Gd_0.2-xYb_xO_3-δ (x?=?0–0.20), proton conducting materials are prepared by the freeze-drying precursor method. The sintering conditions were optimized by adding Zn(NO₃)₂·6H₂O as sintering additive. The materials are thoroughly characterized by different structural and microstructural techniques, including X-ray diffraction, scanning and transmission electron microscopy, and thermogravimetric-differential thermal analysis. The addition of Zn favours the phase formation and densification at lower sintering temperatures; however, it leads to the segregation of a Zn-rich secondary phase, with general formula BaLn₂ZnO₅ (Ln?Y, Gd and Yb), which is identified and quantified for the first time. All samples with Zn as sintering aid exhibit cubic structure; however, the samples without Zn crystallize with orthorhombic or cubic structure, depending on the composition and thermal treatment. The electrical properties are studied by impedance spectroscopy. A deep analysis of the bulk and grain boundary contributions to the conductivity has revealed that the bulk conductivity remains almost unchanged along both series over Yb-doping; however, the grain boundary resistance decreases. The highest conductivity values are found for the intermediate members of both series, BaCe_0.6Zr_0.2Y_0.1Yb_0.1O_3-δ and BaCe_0.6Zr_0.2Gd_0.1Yb₁O_3-δ, with 33 and 28?mS?cm^?1 at 750?°C, respectively.     

An improved algorithm for the three‐fluid‐phase VLLE flash calculation

The VLLE flash is important in water and hydrocarbons mixtures, hydrocarbon and CO₂ rich mixtures, and hydrocarbon methane rich mixtures that are encountered in reservoir performance and recovery studies. A robust VLLE flash algorithm is proposed. The equilibrium and mass balance equations are solved as a constrained minimization problem. An inverse barrier function is used to handle the inequality constrains to solve for the phase fractions. It warrants always arriving to the solution. The challenging cases analyzed showed that the initialization procedure proposed, together with successive substitution iteration in the outer loop, is a good method for a stable VLLE flash algorithm, even near critical points. Whenever the result is in the region outside the three‐phase physical domain, the solution suggests that the system has fewer phases. In one of the cases analyzed, a region with three liquid phases was encountered and the algorithm found two different solutions with positive phase fractions. © 2015 American Institute of Chemical Engineers AIChE J, 61: 3081–3093, 2015     

Morphology,mechanical performance,and nanoindentation behavior of injection molded PC/ABS‐MWCNT nanocomposites

In this work, nanocomposites of polycarbonate/acrylonitrile‐butadiene‐styrene (PC/ABS) with various loads of multiwall carbon nanotubes (MWCNT) are investigated. Material is previously formed by masterbatch dilution approach and further processed by injection molding at various velocities. Microscopic characterization of nanocomposites morphology reveals stronger dependence of MWCNT dispersion on processing parameters at higher nanofiller load. Dispersion of carbon nanotubes at various distances from the injection gate is studied by Raman spectroscopy showing lower deviation at elevated injection velocity. Nanoindentation results that are in agreement with uniaxial tensile testing show a slight decrease of nanocomposites' mechanical performance at 3.0 wt % MWCNT in samples injected at reduced velocity. This is explained by the increase of agglomeration behavior at these conditions. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 42014.     

Understanding web site redesigns in small- and medium-sized enterprises (SMEs): a U.K.-based study on the applicability of e-commerce Stage Models

Despite the efforts of governments and the various support programmes, achievement of advanced stages of e-commerce by small- and medium-sized enterprises (SMEs) is still very low. There have been some attempts to study the dynamic nature of websites, but there is still little research evidence to explain why and how SMEs evolve their web presence. This paper aims to develop a comprehensive classification of drivers for web site redesign based on interviews with various members of staff from SMEs in the U.K. that have recently redesigned their web sites. A sequential mixed-methodological analysis, involving the use of qualitative and quantitative data analysis, was used to develop the classification. This enabled the development of a framework that classified seven main categories of drivers for web site redesign. The drivers identified were: changing business requirements, evolving internet strategies, addressing user needs, maintenance, changing technology, pressure from peers/competitors, and the influence of developers. However, only the first four were found to be significant in the study. The categorisation and the findings suggest a number of key determinants not explicitly addressed by other work. In addition, the findings provide little support for the staged approach to e-commerce progression as few companies reported the implementation of sophisticated internet technology features as a main reason for their web site redesigns. The contributions of this paper are firstly, to provide an instrument to the academic and practitioner communities interested in the topic of web site evolution. Secondly, the categorisation of drivers for redesign and the individual reasons found in this study are expected to provide assistance to SME managers to justify, plan and strategise internet investments realistically and effectively.     

A multiplatform Java wrapper for the BioAPI framework

We present a solution for the development of multiplatform and web-oriented Java applications for biometric authentication based on the BioAPI framework. Our proposal is a single Java Native Interface wrapper that is compatible with the BioAPI instantiations of Windows and Linux/Unix operating systems. Following a study of existing wrappers, we extended the open-source Linux/Unix wrapper to include Windows support. In order to show the utility of this multiplatform Java wrapper, we discuss some of its possible uses and present a real-life application.