Tudor staphylococcal nuclease is an evolutionarily conserved component of the programmed cell death degradome

Programmed cell death (PCD) is executed by proteases, which cleave diverse proteins thus modulating their biochemical and cellular functions. Proteases of the caspase family and hundreds of caspase substrates constitute a major part of the PCD degradome in animals(1,2). Plants lack close homologues of caspases, but instead possess an ancestral family of cysteine proteases, metacaspases(3,4). Although metacaspases are essential for PCD(5-7), their natural substrates remain unknown(4,8). Here we show that metacaspase mcII-Pa cleaves a phylogenetically conserved protein, TSN (Tudor staphylococcal nuclease), during both developmental and stress-induced PCD. TSN knockdown leads to activation of ectopic cell death during reproduction, impairing plant fertility. Surprisingly, human TSN (also known as p100 or SND1), a multifunctional regulator of gene expression(9-15), is cleaved by caspase-3 during apoptosis. This cleavage impairs the ability of TSN to activate mRNA splicing, inhibits its ribonuclease activity and is important for the execution of apoptosis. Our results establish TSN as the first biological substrate of metacaspase and demonstrate that despite the divergence of plants and animals from a common ancestor about one billion years ago and their use of distinct PCD pathways, both have retained a common mechanism to compromise cell viability through the cleavage of the same substrate, TSN.

Transfer of amiprophosmethyl resistance from a Nicotiana plumbaginifolia mutant by somatic hybridisation

Transfer of resistance to the phosphorothioamidate herbicide, amiprophosmethyl (APM), from the P-tubulin mutant of Nicotiana plumbaginifolia to the interspecific N, plumbaginifolia (+) N, sylvestris is and to the intertribal N, plumbaginifolia (+) Atropa belladonna somatic hybrids has been demonstrated. Transfer to the recipient species was accomplished by: (1) symmetric hybridisation and (2) asymmetric hybridisation using gamma-irradiation of donor protoplasts. Cytogenetic analysis confirmed the hybrid origin of the hybrids obtained. It was established that most of them typically inherited no more than three donor chromosomes, although it was possible to obtain symmetric hybrids in the case of symmetric fusion. Immunofluorescent microscopy analysis has shown that protoplasts of the mutant, and of the N. plumbagini-folia (+) N. sylvestris and N. plumbaginifolia (+) A. belladonna hybrids, retained the normal structure of interphase microtubule (MT) arrays and mitotic figures after treatment with 5 mu M APM, whereas MTs of protoplasts of the recipients were destroyed under these conditions. It was also shown that hybrid clones contained an altered beta-tubulin isoform originating from the N. plumbaginifolia mutant. The selected hybrid clones were characterised by cross-resistance to trifluralin, a dinitroaniline herbicide with the same mode of anti-MT action. Some of the somatic hybrids which could flower were fertile. It was established that seeds of some fertile hybrids were able to germinate in the presence of 5 mu M APM. The results obtained thus support the conclusion that the technique of somatic hybridisation, especially asymmetric fusion, can be used to transfer APM resistance from the N. plumbaginifolia mutant to different (related and remote) plant species of the Solanaceae, including important crops.

Parameter studies of fibre laser micro-welding of AISI 316L using Taguchi method

This paper discusses the application of the Taguchi experimental design approach in optimizing the key process parameters for micro-welding of thin AISI 316L foil using the 100W CW fibre laser. A L16 Taguchi experiment was conducted to systematically understand how the power, scanning velocity, focus position, gas flow rate and type of shielding gas affect the bead dimensions. The welds produced in the L16 Taguchi experiment was mainly of austenite cellular-dendrite structure with an average grain size of 5µm. An exact penetration weld with the largest penetration to fusion width ratio was obtained. Among those process parameters, the interaction between power and scanning velocity presented the strongest effect to the penetration to fusion width ratio and the power was found to be the predominantly important factor that drives the interaction with other factors to appreciably affect the bead dimensions.

Potassium canrenoate treatment in paediatric patients: a population pharmacokinetic study using novel dried blood spot sampling

Objective: To characterize the population pharmacokinetics of canrenone following administration of potassium canrenoate (K-canrenoate) in paediatric patients.

Methods: Data were collected prospectively from 37 paediatric patients (median weight 2.9?kg, age range 2 days–0.85 years) who received intravenous K-canrenoate for management of retained fluids, for example in heart failure and chronic lung disease. Dried blood spot (DBS) samples (n?=?213) from these were analysed for canrenone content and the data subjected to pharmacokinetic analysis using nonlinear mixed-effects modelling. Another group of patients (n?=?16) who had 71 matching plasma and DBS samples was analysed separately to compare canrenone pharmacokinetic parameters obtained using the two different matrices.

Results: A one-compartment model best described the DBS data. Significant covariates were weight, postmenstrual age (PMA) and gestational age. The final population models for canrenone clearance (CL/F) and volume of distribution (V/F) in DBS were CL/F (l/h)?=?12.86?×? (WT/70.0)0.75?×?e [0.066?×? (PMA?-?40]) and V/F (l)?=?603.30?×? (WT/70)?×?(GA/40)1.89 where weight is in kilograms. The corresponding values of CL/F and V/F in a patient with a median weight of 2.9?kg are 1.11?l/h and 20.48?l, respectively. Estimated half-life of canrenone based on DBS concentrations was similar to that based on matched plasma concentrations (19.99 and 19.37?h, respectively, in 70?kg patient).

Conclusion: The range of estimated CL/F in DBS for the study population was 0.12–9.62?l/h; hence, bodyweight-based dosage adjustment of K-canrenoate appears necessary. However, a dosing scheme that takes into consideration both weight and age (PMA/gestational age) of paediatric patients seems more appropriate.

Parameter Optimization on Laser Welding of NiTi Wires by Taguchi Method

In this research, a preliminary study was done to find out the initial parameter window to obtain the full-penetrated NiTi weldment. A L27 Taguchi experiment was then carried out to statistically study the effects of the welding parameters and their possible interactions on the weld bead aspect ratio (or penetration over fuse-zone width ratio), and to determine the optimized parameter settings to produce the full-penetrated weldment with desirable aspect ratio. From the statistical results in the Taguchi experiment, the laser mode was found to be the most important factor that substantially affects the aspect ratio. Strong interaction between the power and focus position was found in the Taguchi experiment. The optimized weldment was mainly of columnar dendritic structure in the weld zone (WZ), while the HAZ exhibited equiaxed grain structure. The XRD and DSC results showed that the WZ remained the B2 austenite structure without any precipitates, but with a significant decrease of phase transformation temperatures. The results in the micro-hardness and tensile tests indicated that the mechanical properties of NiTi were decreased to a certain extent after fibre laser welding.

Purification of measles virus with preservation of infectivity and antigenicity

Measles virus Edmonston strain was purified by ultrafiltration followed by two successive sedimentations through sucrose. Purified virus retained infectivity and, when used as an immunogen, elicited high titred antibody to measles antigens by conventional serology. The measles preparations were examined by SDS-PAGE followed by staining. In addition, following PAGE, the purity of these preparations was assessed immunochemically using antisera directed to measles and host cell antigens. The results of these studies demonstrate the utility of the purification method for the preparation of milligram quantities of relatively pure measles virus.

Development of a heterogeneous catalyst for lignocellulosic biomass conversion:Glucose dehydration by metal chlorides in a silica-supported ionic liquid layer

An attempt is made to immobilize the homogeneous metal chloride/EMIMCl catalyst for glucose dehydration to 5-hydroxymethylfurfural. To this end, ionic liquid fragments were grafted to the surface of SBA-15 to generate a heterogenized mimick of the homogeneous reaction medium. Despite a decrease in the surface area, the ordered mesoporous structure of SBA-15 was largely retained. Metal chlorides dispersed in such ionic liquid film are able to convert glucose to HMF with much higher yields as is possible in the aqueous phase. The reactivity order CrCl > AlCl > CuCl > FeCl is similar to the order in the ionic liquid solvent, yet the selectivity are lower. The HMF yield of the most promising CrCl-Im-SBA-15 can be improved by using a HO:DMSO mixture as the reaction medium and a 2-butanol/MIBK extraction layer. Different attempts to decrease metal chloride leaching by using different solvents are described. © 2013 American Institute of Chemical Engineers Environ Prog.

Identification of two focal adhesion targeting sequences in the adapter molecule p130(Cas)

The adapter molecule CAS is localized primarily within focal adhesions in fibroblasts. Because many of the cellular functions attributed to CAS are likely to be dependent on its presence in focal adhesions, this study was undertaken to identify regions of the protein that are involved in its localization. The SH3 domain of CAS, when expressed in isolation from the rest of the protein, was able to target to focal adhesions, whereas a variant containing a point mutation that rendered the SH3 domain unable to associate with FAK remained cytoplasmic. However, in the context of full-length CAS, this mutation did not prevent CAS localization to focal adhesions. Two other variants of CAS that contained deletions of either the SH3 domain alone, or the SH3 domain together with an adjoining proline-rich region, also retained the capacity to localize to focal adhesions. A second focal adhesion targeting region was mapped to the extreme carboxy terminus of CAS. The identification of this second focal adhesion targeting domain in CAS ascribes a previously unknown function to the highly conserved C terminus of CAS. The regulated targeting of CAS to focal adhesions by two independent domains may reflect the important role of CAS within this subcellular compartment.

Considering Just-World Thinking in Counter-Terrorism Cases: Miscarriages of Justice in Northern Ireland

Recent literature has drawn a parallel between the discriminatory application of counterterrorism legislation to the Irish population in the United Kingdom during the Northern Ireland conflict and the targeting of Muslims after September 2001. Less attention has been paid to lessons that can be drawn from judicial decision making in terrorism-related cases stemming from the Northern Ireland conflict. This Article examines Northern Ireland Court of Appeal (“NICA”) jurisprudence on miscarriages of justice in cases regarding counterterrorism offenses. In particular, the Article focuses on cases referred after the 1998 peace agreements in Northern Ireland from the Criminal Cases Review Commission (“CCRC”), a relatively new entity that investigates potential wrongful convictions in England, Wales, and Northern Ireland. Although the NICA’s human rights jurisprudence has developed significantly in recent years, the study of CCRC-referred cases finds that judges have retained confidence in the integrity of the conflict-era counterterrorism system even while acknowledging abuses and procedural irregularities that occurred. This study partially contradicts contentions that judicial deference to the executive recedes in a post-conflict or post-emergency period. Despite a high rate of quashed convictions, the NICA’s decisions suggest that it seeks to limit a large number of referrals and demonstrate a judicial predisposition to defend the justness of the past system’s laws and procedure. This perspective is consistent with what social psychologists have studied as “just-world thinking,” in which objective observers, although motivated by a concern with justice, believe—as a result of cognitive bias—that individuals “got what they deserved.” The Article considers other potential interpretations of the jurisprudence and contends that conservative decision making is particularly dangerous in the politicized realm of counterterrorism and in light of the criminalization of members of suspect communities.

Recent contribution of sediments and fluids to the mantle's volatile budget

Subduction modifies the cycling of Earth's volatile elements. Fluid-rich sediments and hydrated oceanic lithosphere enter the convecting mantle at subduction zones. Some of the sediments and volatile components are released from the subducting slab, promote mantle melting and are returned to the surface by volcanism. The remainder continue into the deeper mantle. Quantification of the fate of these volatiles requires an understanding of both the nature and timing of fluid release and mantle melting(1). Here we analyse the trace element and isotopic geochemistry of fragments of upper mantle rocks that were transported to the surface by volcanic eruptions above the Batan Island subduction zone, Philippines. We find that the mantle fragments exhibit extreme disequilibrium between their U-Th-Ra isotopic ratios, which we interpret to result from the interaction of wet sediment melts and slab-derived fluids with rocks in the overlying mantle wedge. We infer that wet sediments were delivered from the slab to the mantle wedge between 8,000 and 10,000 years ago, whereas aqueous fluids were delivered separately much later. We estimate that about 625 ppm of water is retained in the wedge. A significant volume of water could therefore be delivered to the mantle transition zone at the base of the upper mantle, or even to the deeper mantle.

SMAD inhibition attenuates epithelial to mesenchymal transition by primary keratinocytes in vitro

Epithelial to mesenchymal transition (EMT) is a process whereby epithelial cells undergo transition to a mesenchymal phenotype and contribute directly to fibrotic disease. Recent studies support a role for EMT in cutaneous fibrotic diseases including scleroderma and hypertrophic scarring, though there is limited data on the cytokines and signalling mechanisms regulating cutaneous EMT. We investigated the ability of TGF-β and TNF-α, both over-expressed in cutaneous scleroderma and central mediators of EMT in other epithelial cell types, to induce EMT in primary keratinocytes and studied the signalling mechanisms regulating this process. TGF-β induced EMT in normal human epidermal keratinocytes (NHEK cells) and this process was enhanced by TNF-α. EMT was characterised by changes in morphology, proteome (down-regulation of E-cadherin and Zo-1, and up-regulation of vimentin and fibronectin), MMP secretion and COL1α1 mRNA expression. TGF-β and TNF-α in combination activated SMAD and p38 signalling in NHEK cells. P38 inhibition with SB203580 partially attenuated EMT, whereas SMAD inhibition using SB431542 significantly inhibited EMT and also reversed established EMT. These data highlight the retained plasticity of adult keratinocytes and support further studies of EMT in clinically relevant in vivo models of cutaneous fibrosis, and investigation of SMAD inhibition as a potential therapeutic intervention. This article is protected by copyright. All rights reserved.

Optimisation of Environment-friendly SCFRC mixes use in precast Concrete Industry

The concept of green concrete has been progressively introduced in concrete technology. At the same time, new generations of superplasticisers have become widely available and self-compacting concrete is being increasingly implemented. The aim of this research is to study the impact that different sustainable materials have on both fresh and hardened properties of Self-Compacting Fibre Reinforced Concrete (SCFRC) in order to implement their use in a precast concrete company. Different combinations of cement, mineral additions (active and inert), polypropylene fibres, superplasticisers, and aggregates have been considered. Fresh state performance has been assessed by means of: slump flow test, V-funnel, and J-ring. Concrete compressive strength values at different ages have been retained as representative of the material's performance in its hardened state. All these properties have been correlated with SCFRC proportioning parameters. The importance of interactions between mineral additions and between these and superplasticiser is emphasised, as well as the different consequences of using powders as cement replacement or as mineral additions.

Optimisation of Environment-friendly SCFRC mixes use in precast Concrete Industry (PDF Download Available). Available from: http://www.researchgate.net/publication/263304799_Optimisation_of_Environment-friendly_SCFRC_mixes_use_in_precast_Concrete_Industry [accessed Jun 5, 2015].

L'indicateur EU2020 de suivi de la pauvreté et de l'exclusion: une analyse critique

The reduction of poverty and social exclusion is one of the targets of the European Union's 2020 strategy. The appropriateness and success of such a policy require the choice of relevant indicators that not only highlight poverty gaps between countries but also identify the groups of individuals in each country that need particular attention from social policies. The target retained in the European strategy combines three criteria: people living in households below the monetary poverty threshold, poor people “in terms of standard of living” who live in a situation of severe material deprivation, and those who live in households with very low or zero work intensity. We first show that neither the combination nor the intersection of these three criteria produces an adequate measure of the fight against poverty, or an objective for it. We therefore propose an alternative concept, that of “consistent poverty”, which targets people who simultaneously live below the monetary poverty threshold and above a certain level of material deprivation. The special material deprivation module of the EU-SILC 2009 database allows us to examine two versions of this notion of deprivation: the measurement of “severe” deprivation currently used by the European Union, which adopts a threshold with four items, and an alternative measure of “elementary” material deprivation with a three-item threshold. The intersection between our three-item elementary deprivation criterion and the monetary poverty criterion produces more satisfactory results than those obtained by the European Union approach, in terms of both coherency and profile of the population identified.

A unified view on the effect of fibers and loading on sfrc creep through linear projection to latent structures

Creep of Steel Fiber Reinforced Concrete (SFRC) under flexural loads in the cracked state and to what extent different factors determine creep behaviour are quite understudied topics within the general field of SFRC mechanical properties. A series of prismatic specimens have been produced and subjected to sustained flexural loads. The effect of a number of variables (fiber length and slenderness, fiber content, and concrete compressive strength) has been studied in a comprehensive fashion. Twelve response variables (creep parameters measured at different times) have been retained as descriptive of flexural creep behaviour. Multivariate techniques have been used: the experimental results have been projected to their latent structure by means of Principal Components Analysis (PCA), so that all the information has been reduced to a set of three latent variables. They have been related to the variables considered and statistical significance of their effects on creep behaviour has been assessed. The result is a unified view on the effects of the different variables considered upon creep behaviour: fiber content and fiber slenderness have been detected to clearly modify the effect that load ratio has on flexural creep behaviour.

Enhanced laccase stability through mediator partitioning into hydrophobic ionic liquids

Laccase-mediator systems have numerous potential uses for green oxidations, but their practical use may be limited because the reactive, oxidised mediators deactivate the enzyme. TEMPO, 4-hydroxybenzyl alcohol, phenothiazine and 2-hydroxybiphenyl caused almost complete deactivation of laccase from Trametes versicolor within 24-140 h. By contrast, 18% activity was retained after 188 h in controls without mediator, and 15% in the presence of ABTS. A biphasic reaction system was developed to protect the laccase, by partitioning the mediator into water-immiscible ionic liquids. In the presence of [C mim][AOT], laccase retained 54, 35, 35 and 41% activity after 188 h in the presence of 4-hydroxybenzyl alcohol, phenothiazine and 2-hydroxybiphenyl and ABTS, respectively, whilst 30% activity was retained in the presence of [N][Sac] and TEMPO. The protection against deactivation by the mediators correlated strongly with the distribution coefficients of the mediators between ionic liquids and water. © 2014 The Royal Society of Chemistry.