Functional analysis of a rickettsial OmpA homology domain of Shigella flexneri icsA

Shigella flexneri is a gram-negative bacterium that causes diarrhea and dysentery by invasion and spread through the colonic epithelium. Bacteria spread by assembling actin and other cytoskeletal proteins of the host into "actin tails" at the bacterial pole; actin tail assembly provides the force required to move bacteria through the cell cytoplasm and into adjacent cells. The 120-kDa S. flexneri outer membrane protein IcsA is essential for actin assembly. IcsA is anchored in the outer membrane by a carboxy-terminal domain (the beta domain), such that the amino-terminal 706 amino acid residues (the alpha domain) are exposed on the exterior of the bacillus. The alpha domain is therefore likely to contain the domains that are important to interactions with host factors. We identify and characterize a domain of IcsA within the alpha domain that bears significant sequence similarity to two repeated domains of rickettsial OmpA, which has been implicated in rickettsial actin tail formation. Strains of S. flexneri and Escherichia coli that carry derivatives of IcsA containing deletions within this domain display loss of actin recruitment and increased accessibility to IcsA-specific antibody on the surface of intracytoplasmic bacteria. However, site-directed mutagenesis of charged residues within this domain results in actin assembly that is indistinguishable from that of the wild type, and in vitro competition of a polypeptide of this domain fused to glutathione S-transferase did not alter the motility of the wild-type construct. Taken together, our data suggest that the rickettsial homology domain of IcsA is required for the proper conformation of IcsA and that its disruption leads to loss of interactions of other IcsA domains within the amino terminus with host cytoskeletal proteins.     

Factors affecting the potentiometric response of all-solid-state solvent polymeric membrane calcium-selective electrode for low-level measurements

An all-solid-state calcium-selective electrode was constructed with poly(pyrrole) solid-contact doped with calcium complexing ligand Tiron. The potentiometric response of this sensor can have a linear range down to 10(-)(9) M with a slope close to Nernstian and detection limit equal to 10(-)(9.6). The effects of pH and the activity of the interfering ion in the conditioning solution on the potentiometric behavior of the constructed sensors were examined. Potential stability, reproducibility, and impedance studies were performed. The selectivity of the constructed electrode is better than that of the conventional calcium-selective electrode with internal filling solution of 10(-)(2) M CaCl(2) and comparable to that of the best liquid-contact electrodes.     

Strong Negation and Equivalence in the Safe Belief Semantics

The safe belief semantics uses intermediate logics to definean extension of answer sets to all propositional formulas, butonly considering one kind of negation. In this work we extendsafe beliefs adding the strong negation connective. The mainfeature of our extension is that strong negation can occur beforeany formula, and not only at the atomic level. We give resultsconcerning the relation between strong negation extensions ofintermediate logics and safe beliefs and consider the way inwhich strong negation can be eliminated from any formula whilepreserving its semantics. We also propose two new notions ofequivalence: substitution equivalence and contextualized equivalence.We prove that they are both more general than strong equivalenceand, for propositional formulas where strong negation may occurat the non-atomic level, substitution equivalence captures anotion of equivalence that cannot be captured by strong equivalencealone.     

Impact of including land-use change emissions from biofuels on meeting GHG emissions reduction targets: the example of Ireland

The greenhouse gases (GHG) emissions from land-use change are of particular concern for land-based biofuels. Emissions avoided by substituting fossil fuels with biofuels may be offset by emissions from direct and indirect land-use changes (LUC). There is an urgent need to investigate what impact land-use change emissions may have on the expansion of bioenergy and biofuels, in the context of EU mitigation policies. This paper focuses on Ireland, which faces a number of challenges in delivering its renewable energy and GHG reduction targets. The Irish TIMES energy systems model was used to assess the impact of a range of land-use change emissions’ levels on the evolution of Ireland’s low-carbon energy system. A reference scenario was developed where LUC is ignored and Ireland achieves a least-cost low-carbon energy system by 2050. If high indirect land-use change (ILUC) emissions are included, this results in a decrease by 30 % in bioenergy and a 68 % increase in marginal abatement costs by 2050. Hydrogen is used instead of bioenergy in the freight sector in this scenario, while private cars are fuelled by renewable electricity. If GHG emissions from ILUC were considered less severe, indigenous grass biomethane becomes the key biofuel representing 31 % of total bioenergy consumption. This is in line with recent research in Ireland of the key role that grass biomethane can play.     

Chemical Composition of Seed Oils Recovered from Different Pear (Pyrus communis L.) Cultivars

Lipophilic bioactive compounds in oils recovered from the seeds of eight pear (Pyrus communis L.) cultivars were studied. Oil yield in pear seeds ranged between 16.3 and 31.5 % (w/w) dw. The main fatty acids were palmitic acid (6.13–8.52 %), oleic acid (27.39–38.17 %) and linoleic acid (50.73–63.78 %), all three representing 96–99 % of the total detected fatty acids. The range of total tocochromanols was between 120.5 and 216.1 mg/100 g of oil. Independent of the cultivar, the γ‐tocopherol was the main tocochromanol and constituted approximately 88 %. The contents of the carotenoids and squalene were between 0.69–2.99 and 25.5–40.8 mg/100 g of oil, respectively. The β‐sitosterol constituted 83.4–87.6 % of total sterols contents, which ranged between 276.4 and 600.1 mg/100 g of oil. Three significant correlations were found between oil yield and total contents of sterols (r = ?0.893), tocochromanols (r = ?0.955) and carotenoids (r = ?0.685) in pear seed oils.     

In‐vitro study on the efficacy of tannin fractions of edible nuts as antioxidants

Tannin fractions were isolated from crude acetonic extracts of defatted walnut, hazelnut and almond kernels using Sephadex LH‐20 column chromatography. The obtained material was characterized by content of total phenolics and electrophoretic separations using capillary zone electrophoresis (CZE). The antioxidant activities of the tannin fractions were analyzed by several methods: DPPH and ABTS assays, photochemiluminescence (PCL) method, as well as in two lipid model systems: emulsion with β‐carotene‐linoleic acid and L ‐α‐lecithin liposomes. The contents of total phenolics in the tannin fractions of walnuts, hazelnuts and almonds were 550, 329 and 83 mg catechin eq/g, respectively. The electrophoretic profiles of hazelnut and almond tannin fractions were similar, in contrast to the walnut profile. All analyzed fractions exhibited strong antioxidant properties. The antioxidant capacity of lipid‐soluble (ACL) compounds determined by PCL method was the highest for the fraction isolated from walnuts – 7.35 mmol Trolox eq/g. The DPPH radical and the ABTS radical cation were scavenged by the walnut tannin fraction with a higher efficacy than by the two other fractions. EC₅₀ values of the DPPH method were 1.8 times higher for the hazelnut fraction and 2.3 times higher for the almond fraction when compared to the walnut tannins. In turn, the total antioxidant activity values were 8.17, 2.82 and 1.98 mmol Trolox eq/g for the walnut, hazelnut and almond fractions, respectively. On the other hand, in both lipid models applied, lower antioxidant activity of walnut tannins than of hazelnut tannins was noted. The antioxidant effect of almond tannins was weaker or similar than that of walnut tannins in the β‐carotene‐linoleic acid emulsion and the L ‐α‐lecithin liposomal system, respectively.     

In-vessel composting for utilizing of municipal sewage-sludge

The amount of sludge generated from wastewater treatment plants has been increasing year-by-year as the sewerage service area expanded. The composting process is considered to be one of the best management methods to stabilize different organic wastes. The main goal of this investigation was to find the optimal ratio of the quantity of the organic waste to the quantity of the sewage sludge. The process of composting was carried out in the Horstmann bioreactor (pilot scale) for 2–3 weeks and then in a compost pile for 6–12 weeks. Seven experiments were performed, with different ratios of the quantity of organic wastes to the quantity of sewage sludge. The results enabled an estimation of the quality of compost and its usefulness and also the usefulness of the bioreactor to be deduced.