Expression, purification and structural analysis of the Pyrococcus abyssi RNA binding protein PAB1135

<p>Abstract</p> <p>Background</p> <p>The gene coding for the uncharacterized protein PAB1135 in the archaeon <it>Pyrococcus abyssi </it>is in the same operon as the ribonuclease P (RNase P) subunit Rpp30.</p> <p>Findings</p> <p>Here we report the expression, purification and structural analysis of PAB1135. We analyzed the interaction of PAB1135 with RNA and show that it binds efficiently double-stranded RNAs in a non-sequence specific manner. We also performed molecular modeling of the PAB1135 structure using the crystal structure of the protein Af2318 from <it>Archaeoglobus fulgidus </it>(<ext-link ext-link-id="2OGK" ext-link-type="pdb">2OGK</ext-link>) as the template.</p> <p>Conclusions</p> <p>Comparison of this model has lead to the identification of a region in PAB1135 that could be involved in recognizing double-stranded RNA.</p

A P-type ATPase importer that discriminates between essential and toxic transition metals

Transition metals, although being essential cofactors in many physiological processes, are toxic at elevated concentrations. Among the membrane-embedded transport proteins that maintain appropriate intracellular levels of transition metals are ATP-driven pumps belonging to the P-type ATPase superfamily. These metal transporters may be differentiated according to their substrate specificities, where the majority of pumps can extrude either silver and copper or zinc, cadmium, and lead. In the present report, we have established the substrate specificities of nine previously uncharacterized prokaryotic transition-metal P-type ATPases. We find that all of the newly identified exporters indeed fall into one of the two above-mentioned categories. In addition to these exporters, one importer, Pseudomonas aeruginosa Q9I147, was also identified. This protein, designated HmtA (heavy metal transporter A), exhibited a different substrate recognition profile from the exporters. In vivo metal susceptibility assays, intracellular metal measurements, and transport experiments all suggest that HmtA mediates the uptake of copper and zinc but not of silver, mercury, or cadmium. The substrate selectivity of this importer ensures the high-affinity uptake of essential metals, while avoiding intracellular contamination by their toxic counterparts

Good education policy making: data-informed but values-driven

In this article, based on their book Educational Goods: Values, Evidence and Decision Making, Harry Brighouse, Helen Ladd, Susanna Loeb, and Adam Swift encourage education decision makers to give careful thought to the values that underlie the data they collect and use to inform policy. Rather than basing decisions entirely on what improves academic achievement, the authors call for attention to a wider array of values, which they call educational goods. These include the capacities to function in the labor market, to participate effectively in the democratic process, to make autonomous judgments about key life decisions such as occupation or religion, to develop healthy interpersonal relationships, to seek personal fulfilment, and to treat others with respect and dignity. Thinking in terms of these values can broaden the conversation about education priorities and bring clarity to decisions involving trade-offs and conflicting aims

Assessment for learning : a model for the development of a child’s self competence in the early years of education

In recent years policy documents, curricula and other educational initiatives have promoted a pedagogy founded on the concept of independent learning. This is broadly defined as ‘having the belief in yourself to think through learning activities, problems or challenges, make decisions about your learning and act upon those decisions (Blandford and Knowles, 2009:336). The central role of Assessment for Learning (AfL) in this process is often overlooked in practice. By considering the findings from a small scale research study this article addresses the central role of the teacher /practitioner in developing effective AfL in the early years classroom (3-5 years)

Cosmological constraints on the magnification bias on sub-millimetre galaxies after large-scale bias corrections

Context. The study of the magnification bias produced on high-redshift sub-millimetre galaxies by foreground galaxies through the analysis of the cross-correlation function was recently demonstrated as an interesting independent alternative to the weak-lensing shear as a cosmological probe. Aims. In the case of the proposed observable, most of the cosmological constraints mainly depend on the largest angular separation measurements. Therefore, we aim to study and correct the main large-scale biases that aect foreground and background galaxy samples to produce a robust estimation of the cross-correlation function. Then we analyse the corrected signal to derive updated cosmological constraints. Methods. We measured the large-scale, bias-corrected cross-correlation functions using a background sample of H-ATLAS galaxies with photometric redshifts &gt;1.2 and two dierent foreground samples (GAMA galaxies with spectroscopic redshifts or SDSS galaxies with photometric ones, both in the range 0.2 &lt; z &lt; 0.8). These measurements are modelled using the traditional halo model description that depends on both halo occupation distribution and cosmological parameters. We then estimated these parameters by performing a Markov chain Monte Carlo under multiple scenarios to study the performance of this observable and how to improve its results. Results. After the large-scale bias corrections, we obtain only minor improvements with respect to the previous magnification bias results, mainly confirming their conclusions: a lower bound on m &gt; 0:22 at 95% CL and an upper bound 8 &lt; 0:97 at 95% CL (results from the zspec sample). Neither the much higher surface density of the foreground photometric sample nor the assumption of Gaussian priors for the remaining unconstrained parameters significantly improve the derived constraints. However, by combining both foreground samples into a simplified tomographic analysis, we were able to obtain interesting constraints on the m8 plane as follows: m = 0:50+0:14 0:20 and 8 = 0:75+0:07 0:10 at 68% C

The impact of pumped water from a de-watered Magnesian limestone quarry on an adjacent wetland: Thrislington, County Durham, UK

Although quarrying is often cited as a potential threat to wetland systems, there is a lack of relevant, quantitative case studies in the literature. The impact of pumped groundwater discharged from a quarry into a wetland area was assessed relative to reference conditions in an adjacent fen wetland that receives only natural runoff. Analysis of vegetation patterns at the quarry wetland site, using Detrended Correspondence Analysis and the species indicator values of Ellenberg, revealed a clear disparity between community transitions in the quarry wetland and the reference site. Limited establishment of moisture-sensitive taxa, the preferential proliferation of robust wetland species and an overall shift towards lower species diversity in the quarry wetland were explicable primarily by the physico-chemical environment created by quarry dewatering. This encompassed high pH (up to 12.8), sediment-rich effluent creating a nutrient-poor substrate with poor moisture retention in the quarry wetland, and large fluctuations in water levels. © 2005 Elsevier Ltd. All rights reserved

Insecticidal effects of plant extracts on immature whitefly Bemisia tabaci Genn. (Hemiptera: Aleyroideae)

Background: The whitefly ( Bemisia tabaci Genn.) is a widely distributed and highly harmful plant pest species. The management of B. tabaci has been typically carried out by chemical pesticides. In the last decade however, there has been an increasing interest in natural products, particularly those of plant origin, to control this pest species. In the present work, aqueous and ethanolic extracts of native plants from the flora of the Yucat\ue1n peninsula ( Acalypha gaumeri , Annona squamosa , Carlowrightia myriantha, Petiveria alliaceae and Trichilia arborea) and the introduced plant Azadirachta indica were collected and evaluated for insecticidal activity against eggs and nymphs Bemisia tabaci. Results: Most of the aqueous and ethanolic extracts showed high insecticidal effects on B. tabaci eggs. The lowest LC50 values were recorded in the aqueous extracts of A. gaumeri (0.39% w/v), A. squamosa (0.36% w/v), P. alliaceae (0.42% w/v) and A. indica (0.30% /v), as well as in the ethanolic extracts of P. alliaceae (2.09 mg mL-1) and T. arborea (2.14 mg mL-1). On the other hand, B. tabaci nymphs were not affected by the aqueous extracts, but were highly sensitive to the ethanolic extracts of the tested plants. The lowest LC50 values were recorded in the ethanolic extracts of P. alliaceae (1.27 mg ml-1) and T. arborea (1.61 mg mL-1). The GC-MS analysis showed that phytol was the major component of the ethanolic extract of P. alliaceae and fatty acids were the major components of ethanolic extract of T. arborea. Conclusions: Overall, results suggest that ethanolic extracts of P. alliaceae and T. arborea leaves showed the highest insecticidal effects on eggs and nymphs B. tabaci. The extracts from P. alliaceae and T. arborea are good candidates to be developed as sources of natural insecticides for the management of immature B. tabaci since their effects were comparable with that showed by the extracts of A. indica, a well-known plant species for its insecticidal activity

Cytotoxic Acetogenins from the Roots of Annona purpurea

Annona purpurea, known in Mexico as "cabeza de negro" or "ilama", belongs to the Annonaceae family. Its roots are employed in folk medicine in several regions of Mexico. Taking that information into account, a chemical and biological analysis of the components present in the roots of this species was proposed. Our results demonstrated that the dichloromethane (DCM) extract was exclusively constituted by a mixture of five new acetogenins named annopurpuricins A-E (1-5). These compounds have an aliphatic chain of 37 carbons with a terminal \u3b1,\u3b2 unsaturated \u3b3-lactone. Compounds 1 and 2 belong to the adjacent bis-THF (tetrahydrofuran) \u3b1-monohydroxylated type, while compounds 3 and 4 belong to the adjacent bis-THF \u3b1,\u3b1'-dihydroxylated type; only compound 5 possesses a bis-epoxide system. Complete structure analysis was carried out by spectroscopy and chemical methods. All compounds were evaluated for their antiproliferative activity on three human tumor cell lines (MSTO-211H, HeLa and HepG2). Compounds 1-4 inhibited significantly the growth of HeLa and HepG2 cells, showing GI50 values in the low/subnanomolar range, while 5 was completely ineffective under the tested conditions. The investigation of the mechanism of action responsible for cytotoxicity revealed for the most interesting compound 1 the ability to block the complex I activity on isolated rat liver mitochondria (RLM)

Roles of a conserved proline in the internal fusion peptide of Ebola glycoprotein

The structural determinants underlying the functionality of viral internal fusion peptides (IFPs) are not well understood. We have compared EBOwt (GAAIGLAWIPY- FGPAAE), representing the IFP of the Ebola fusion protein GP, and EBOmut (GAAIGLAWIPYFGRAAE) derived from a non-functional mutant with conserved Pro537 substituted by Arg. P537R substitution did not abrogate peptide-membrane association, but interfered with the ability to induce bilayer destabilization. Structural determinations suggest that Pro537 is required to preserve a membrane-perturbing local conformation in apolar environments