Mutational analysis of active-site residues in the Mycobacterium leprae RecA intein, a LAGLIDADG homing endonuclease: Asp(122) and Asp(193) are crucial to the double-stranded DNA cleavage activity whereas Asp(218) is not

Mycobacterium leprae recA harbors an in-frame insertion sequence that encodes an intein homing endonuclease (PI-MleI). Most inteins (intein endonucleases) possess two conserved LAGLIDADG (DOD) motifs at their ctive center. A common feature of LAGLIDADG-type homing endonucleases is that they recognize and cleave the same or very similar DNA sequences. However, PI-MleI is distinctive from other members of the family of LAGLIDADG-type HEases for its modular structure with functionally separable domains for DNA-binding and cleavage, each with distinct sequence preferences. Sequence alignment analyses of PI-MleI revealed three putative LAGLIDADG motifs; however, there is conflicting bioinformatics data in regard to their identity and specific location within the intein polypeptide. To resolve this conflict and to determine the active-site residues essential for DNA target site recognition and double-stranded DNA cleavage, we performed site-directed mutagenesis of presumptive catalytic residues in the LAGLIDADG motifs. Analysis of target DNA recognition and kinetic parameters of the wild-type PI-MleI and its variants disclosed that the two amino acid residues, Asp(122) (in Block C) and Asp(193) (in functional Block E), are crucial to the double-stranded DNA endonuclease activity, whereas Asp(218) (in pseudo-Block E) is not. However, despite the reduced catalytic activity, the PI-MleI variants, like the wild-type PI-MleI, generated a footprint of the same length around the insertion site. The D122T variant showed significantly reduced catalytic activity, and D122A and D193A mutations although failed to affect their DNA-binding affinities, but abolished the double-stranded DNA cleavage activity. On the other hand, D122C variant showed approximately twofold higher double-stranded DNA cleavage activity, compared with the wild-type PI-MleI. These results provide compelling evidence that Asp(122) and Asp(193) in DOD motif I and II, respectively, are bona fide active-site residues essential for DNA cleavage activity. The implications of these results are discussed in this report.

An adiabatic model for a two-stage double-inlet pulse tube refrigerator

A numerical modelling technique for predicting the detailed performance of a double-inlet type two-stage pulse tube refrigerator has been developed. The pressure variations in the compressor, pulse tube, and reservoir were derived, assuming the stroke volume variation of the compressor to be sinusoidal. The relationships of mass flowrates, volume flowrates, and temperature as a function of time and position were developed. The predicted refrigeration powers are calculated by considering the effect of void volumes and the phase shift between pressure and mass flowrate. These results are compared with the experimental results of a specific pulse tube refrigerator configuration and an existing theoretical model. The analysis shows that the theoretical predictions are in good agreement with each other.

Experimental investigation on lateral impact response of concrete-filled double-skin tube columns using horizontal-impact-testing system

This paper presents an experimental investigation on the lateral impact performance of axially loaded concrete-filled double-skin tube (CFDST) columns. These columns have desirable structural and constructional properties and have been used as columns in building, legs of off shore platforms and as bridge piers. Since they could be vulnerable to impact from passing vessels or vehicles, it is necessary to understand their behaviour under lateral impact loads. With this in mind, an experimental method employing an innovative instrumented horizontal impact testing system (HITS) was developed to apply lateral impact loads whilst the column maintained a static axial pre-loading to examine the failure mechanism and key response parameters of the column. These included the time histories of impact force, reaction forces, global lateral deflection and permanent local buckling profile. Eight full scale columns were tested for key parameters including the axial load level and impact location. Based on the test data, the failure mode, peak impact force, impact duration, peak reaction forces, reaction force duration, column maximum and residual global deflections and column local buckling length, depth and width under varying conditions are analysed and discussed. It is evident that the innovative HITS can successfully test structural columns under the combination of axial pre-loading and impact loading. The findings on the lateral impact response of the CFDST columns can serve as a benchmark reference for their future analysis and design.

Critical test for Altshuler-Aronov theory: Evolution of the density of states singularity in double perovskite Sr2FeMoO6 with controlled disorder

With high-resolution photoemission spectroscopy measurements, the density of states (DOS) near the Fermi level (E-F) of double perovskite Sr2FeMoO6 having different degrees of Fe/Mo antisite disorder has been investigated with varying temperature. The DOS near E-F showed a systematic depletion with increasing degree of disorder, and recovered with increasing temperature. Altshuler-Aronov (AA) theory of disordered metals well explains the dependences of the experimental results. Scaling analysis of the spectra provides experimental indication for the functional form of the AA DOS singularity.

Investigation of the local Fe magnetic moments at the grain boundaries of the Ca2FeReO6 double perovskite

The local Fe ferromagnetic (FM) moment at the grain boundaries of a ceramic sample of Ca2FeReO6 double perovskite was investigated by means of x-ray magnetic circular dichroism spectroscopy at the Fe L-2,L-3 edges and compared to the overall bulk magnetization. We found that, at the grain boundaries, the Fe FM moments at H=5 T are much smaller than expected and that the MxH curve is harder than in the bulk magnetization. These results suggest a larger degree of Fe/Re antisite disorder at the grain boundaries of this sample, shedding light into the intriguing nonmetallic resistivity behavior despite the reported presence of free carriers. (c) 2007 American Institute of Physics.

Concrete-filled double skin tube columns subjected to lateral impact loading: Experimental and numerical study

This research treats the lateral impact behaviour of composite columns, which find increasing use as bridge piers and building columns. It offers (1) innovative experimental methods for testing structural columns, (2) dynamic computer simulation techniques as a viable tool in analysis and design of such columns and (3) significant new information on their performance which can be used in design. The research outcomes will enable to protect lives and properties against the risk of vehicular impacts caused either accidentally or intentionally.

IMPOSE (IMProving Outcomes after Sepsis)—the effect of a multidisciplinary follow-up service on health-related quality of life in patients postsepsis syndromes—a double-blinded randomised controlled trial: protocol

Introduction Patients post sepsis syndromes have a poor quality of life and a high rate of recurring illness or mortality. Follow-up clinics have been instituted for patients postgeneral intensive care but evidence is sparse, and there has been no clinic specifically for survivors of sepsis. The aim of this trial is to investigate if targeted screening and appropriate intervention to these patients can result in an improved quality of life (Short Form 36 health survey (SF36V.2)), decreased mortality in the first 12 months, decreased readmission to hospital and/or decreased use of health resources. Methods and analysis 204 patients postsepsis syndromes will be randomised to one of the two groups. The intervention group will attend an outpatient clinic two monthly for 6 months and receive screening and targeted intervention. The usual care group will remain under the care of their physician. To analyse the results, a baseline comparison will be carried out between each group. Generalised estimating equations will compare the SF36 domain scores between groups and across time points. Mortality will be compared between groups using a Cox proportional hazards (time until death) analysis. Time to first readmission will be compared between groups by a survival analysis. Healthcare costs will be compared between groups using a generalised linear model. Economic (health resource) evaluation will be a within-trial incremental cost utility analysis with a societal perspective. Ethics and dissemination Ethical approval has been granted by the Royal Brisbane and Women’s Hospital Human Research Ethics Committee (HREC; HREC/13/QRBW/17), The University of Queensland HREC (2013000543), Griffith University (RHS/08/14/HREC) and the Australian Government Department of Health (26/2013). The results of this study will be submitted to peer-reviewed intensive care journals and presented at national and international intensive care and/or rehabilitation conferences.

From the {Cu(μ2-S)N}4 butterfly architecture to the {Cu(μ3-S)N}12 double wheel

Copper(I) complexes with {Cu(μ2-S)N}4 and {Cu(μ3-S)N}12 core portions of butterfly-shaped or double wheel architectures have been isolated in the reaction of Cu(I) with the Schiff base ligand C6H4(CHNC6H4S)2, aiso-abtâ, under different conditions. View the MathML source containing the tetranuclear electroneutral complex View the MathML source is formed by the reaction of CuI in acetonitrilic solution and recrystallization from DMF, whereas View the MathML source containing dodecanuclear View the MathML source wheels is accessible starting from CuBF4. Complexes 2 and 4 represent the first examples of cyclic complexes with the same overall stoichiometry but different ring sizes. The ligand induces two different coordination environments around copper(I) by switching between μ2- and μ3-sulfur bridging modes.

Experimental study on behaviour of concrete filled double skin tube columns under lateral impact loading

This paper presents an experimental investigation on the lateral impact response of axially loaded concrete filled double skin tube (CFDST) columns. A total of four test series are being conducted at Queensland University of Technology using a novel horizontal impact-testing rig. The test results reported in this paper are from the first test series, where the columns are pinned at both ends and impacted at mid-span. In the next three series, effects of support conditions, impact location and repeated impact will be treated. The main objectives of the current paper are to describe the innovative testing procedure and provide some insight into the lateral impact behavior and failure of simply supported axially pre-loaded CFDST columns. The results include time histories of impact forces, reaction forces, axial force and global lateral deflection. Based on the test data, the failure mode, peak impact force, peak reaction forces, maximum deflection and residual deflection, with and without axial load, are analyzed and discussed. The findings of this study will serve as a benchmark reference for future analysis and design of CFDST columns.

Tendency toward crossover of the effective susceptibility exponent from its doubled Ising value to its doubled mean-field value near a double critical point

The critical behavior of osmotic susceptibility in an aqueous electrolyte mixture 1-propanol (1P)+water (W)+potassium chloride is reported. This mixture exhibits re-entrant phase transitions and has a nearly parabolic critical line with its apex representing a double critical point (DCP). The behavior of the susceptibility exponent is deduced from static light-scattering measurements, on approaching the lower critical solution temperatures (TL’s) along different experimental paths (by varying t) in the one-phase region. The light-scattering data analysis substantiates the existence of a nonmonotonic crossover behavior of the susceptibility exponent in this mixture. For the TL far away from the DCP, the effective susceptibility exponent γeff as a function of t displays a nonmonotonic crossover from its single limit three-dimensional (3D)-Ising value ( ∼ 1.24) toward its mean-field value with increase in t. While for that closest to the DCP, γeff displays a sharp, nonmonotonic crossover from its nearly doubled 3D-Ising value toward its nearly doubled mean-field value with increase in t. The renormalized Ising regime extends over a relatively larger t range for the TL closest to the DCP, and a trend toward shrinkage in the renormalized Ising regime is observed as TL shifts away from the DCP. Nevertheless, the crossover to the mean-field limit extends well beyond t>10−2 for the TL’s studied. The observed crossover behavior is attributed to the presence of strong ion-induced clustering in this mixture, as revealed by various structure probing techniques. As far as the critical behavior in complex or associating mixtures with special critical points (like the DCP) is concerned, our results indicate that the influence of the DCP on the critical behavior must be taken into account not only on the renormalization of the critical exponent but also on the range of the Ising regime, which can shrink with decrease in the influence of the DCP and with the extent of structuring in the system. The utility of the field variable tUL in analyzing re-entrant phase transitions is demonstrated. The effective susceptibility exponent as a function of tUL displays a nonmonotonic crossover from its asymptotic 3D-Ising value toward a value slightly lower than its nonasymptotic mean-field value of 1. This behavior in the nonasymptotic, high tUL region is interpreted in terms of the possibility of a nonmonotonic crossover to the mean-field value from lower values, as foreseen earlier in micellar systems.

On gravitational fields of stars in f(R) gravity theories

Einstein's general relativity is a classical theory of gravitation: it is a postulate on the coupling between the four-dimensional, continuos spacetime and the matter fields in the universe, and it yields their dynamical evolution. It is believed that general relativity must be replaced by a quantum theory of gravity at least at extremely high energies of the early universe and at regions of strong curvature of spacetime, cf. black holes. Various attempts to quantize gravity, including conceptually new models such as string theory, have suggested that modification to general relativity might show up even at lower energy scales. On the other hand, also the late time acceleration of the expansion of the universe, known as the dark energy problem, might originate from new gravitational physics. Thus, although there has been no direct experimental evidence contradicting general relativity so far - on the contrary, it has passed a variety of observational tests - it is a question worth asking, why should the effective theory of gravity be of the exact form of general relativity? If general relativity is modified, how do the predictions of the theory change? Furthermore, how far can we go with the changes before we are face with contradictions with the experiments? Along with the changes, could there be new phenomena, which we could measure to find hints of the form of the quantum theory of gravity? This thesis is on a class of modified gravity theories called f(R) models, and in particular on the effects of changing the theory of gravity on stellar solutions. It is discussed how experimental constraints from the measurements in the Solar System restrict the form of f(R) theories. Moreover, it is shown that models, which do not differ from general relativity at the weak field scale of the Solar System, can produce very different predictions for dense stars like neutron stars. Due to the nature of f(R) models, the role of independent connection of the spacetime is emphasized throughout the thesis.

Self-assembly of a nanoscopic platinum(II) double square cage

Self-assembly of a rigid tripyridyl linker with a bidentate 90 degrees Pt(II) acceptor yielded a somewhat unusual double square cage, representing the first example of Pt(II) cage of such shape. Multinuclear NMR as well as single-crystal structure analysis characterized the cage.

Self-assembly of a nanoscopic platinum(II) double square cage

Self-assembly of a rigid tripyridyl linker with a bidentate 90 degrees Pt(II) acceptor yielded a somewhat unusual double square cage, representing the first example of Pt(II) cage of such shape. Multinuclear NMR as well as single-crystal structure analysis characterized the cage.

Graphene-like two-dimensional ionic boron with double dirac cones at ambient condition

Recently, partially ionic boron (γ-B28) has been predicted and observed in pure boron, in bulk phase and controlled by pressure [Nature, 457 (2009) 863]. By using ab initio evolutionary structure search, we report the prediction of ionic boron at a reduced dimension and ambient pressure, namely, the two-dimensional (2D) ionic boron. This 2D boron structure consists of graphene-like plane and B2 atom pairs, with the P6/mmm space group and 6 atoms in the unit cell, and has lower energy than the previously reported α-sheet structure and its analogues. Its dynamical and thermal stability are confirmed by the phonon-spectrum and ab initio molecular dynamics simulation. In addition, this phase exhibits double Dirac cones with massless Dirac fermions due to the significant charge transfer between the graphene-like plane and B2 pair that enhances the energetic stability of the P6/mmm boron. A Fermi velocity (vf) as high as 2.3 x 106 m/s, which is even higher than that of graphene (0.82 x 106 m/s), is predicted for the P6/mmm boron. The present work is the first report of the 2D ionic boron at atmospheric pressure. The unique electronic structure renders the 2D ionic boron a promising 2D material for applications in nanoelectronics.

Interlayer structure of iodide intercalated layered double hydroxides (LDHs)

The iodide-containing layered double hydroxides (LDHs) of Mg and Zn with AI crystallize by the inclusion of extensive positional disorder of I- ions in the interlayer region. I- ion given its poor charge to size ratio can neither screen effectively the positive charge nor participate in H-bonding with the metal hydroxide layers. Thereby the I- ions are not stabilized in sites close to the seat of positive charge of the metal hydroxide layers (6c), nor in sites that facilitate H-bonding (3b or 18h). On the other hand, OH- from water can do both and effectively displaces I- from the interlayer. (C) 2010 Elsevier Inc. All rights reserved.