996 resultados para DEAD-box RNA Helicases


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The Australian Curriculum marks national reforms in social science education, first with the return to the disciplines of history and geography and second, through a new approach to interdisciplinary learning. This paper raises the question of whether the promise of interdisciplinary learning can be realised in the middle years of schooling if teachers have to teach history as a discipline rather than within an over-arching integrated curriculum framework. The paper explores the national blueprints and considers the national history curriculum in light of theories of teachers’ knowledge and middle school education. Evidence from teacher interviews indicates that historical understanding can be achieved through integrated frameworks to meet the goals of middle schooling.

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The QUT Team developed an idea for a new residential housing typology that is appropriate for sites where the best views are in the opposing direction to the preferable climatic orientation. The interlocking configuration creates a double height external living space in every apartment, creating further opportunities for cross ventilation and natural daylight. Unlike conventional double loaded housing typologies, the interlocking configuration only requires a continuous public circulation corridor every second level. The cores that service this corridor are separated to either end of the tower and open areas. The configuration of the interlocking apartments creates an interesting composition of solid and void when viewed externally. This undulating facade petternation assists in articulating the large building mass. The project was evaluated by independent consultants and found to be cost effective, and at the same time delivering energy efficient high density liveability. The project was presented to a meeting of the Australian Council on Tall Buildings seminar on 15 September 2010.

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‘Top Ten Box Office Blockbusters in Dollars’, is an ongoing series of works that represent the production budgets and worldwide gross profits of the top ten grossing films of all time. By displaying this data on top of the full running time of each blockbuster, the viewer’s attention is drawn back and forth between the amassing dollar figures, and the original film’s highly polished presentation. In doing so, the work aims to provide a new opportunity to enjoy these immensely popular films with a new sense of value. The exhibition was selected for the Artistic Program at MetroArts, Brisbane in 2010

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There is an intimate interconnectivity between policy guidelines defining reform and the delineation of what research methods would be subsequently applied to determine reform success. Research is guided as much by the metaphors describing it as by the ensuing empirical definition of actions of results obtained from it. In a call for different reform policy metaphors Lumby and English (2010) note, “The primary responsibility for the parlous state of education... lies with the policy makers that have racked our schools with reductive and dehumanizing processes, following the metaphors of market efficiency, and leadership models based on accounting and the characteristics of machine bureaucracy” (p. 127)

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The Web Service Business Process Execution Language (BPEL) lacks any standard graphical notation. Various efforts have been undertaken to visualize BPEL using the Business Process Modelling Notation (BPMN). Although this is straightforward for the majority of concepts, it is tricky for the full BPEL standard, partly due to the insufficiently specified BPMN execution semantics. The upcoming BPMN 2.0 revision will provide this clear semantics. In this paper, we show how the dead path elimination (DPE) capabilities of BPEL can be expressed with this new semantics and discuss the limitations. We provide a generic formal definition of DPE and discuss resulting control flow requirements independent of specific process description languages.

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This is a professional practice paper for Psychology practitioners to reflect on their skills and therapeutic practices. A Master- practitioner model or Artizan - apprentice analogy is used to understand the development of a practicing psychologist from his/her "salad days" (when we are green [Shakespeare- Anthony and Cleopatra]) to our Autumn years in the profession.

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Evidence for a two-metal ion mechanism for cleavage of the HH16 hammerhead ribozyme is provided by monitoring the rate of cleavage of the RNA substrate as a function of La3+ concentration in the presence of a constant concentration of Mg2+. We show that a bell-shaped curve of cleavage activation is obtained as La3+ is added in micromolar concentrations in the presence of 8 mM Mg2+, with a maximal rate of cleavage being attained in the presence of 3 microM La3+. These results show that two-metal ion binding sites on the ribozyme regulate the rate of the cleavage reaction and, on the basis of earlier estimates of the Kd values for Mg2+ of 3.5 mM and > 50 mM, that these sites bind La3+ with estimated Kd values of 0.9 and > 37.5 microM, respectively. Furthermore, given the very different effects of these metal ions at the two binding sites, with displacement of Mg2+ by La3+ at the stronger (relative to Mg2+) binding site activating catalysis and displacement of Mg2+ by La3+ at the weaker (relative to Mg2+) (relative to Mg2+) binding site inhibiting catalysis, we show that the metal ions at these two sites play very different roles. We argue that the metal ion at binding site 1 coordinates the attacking 2'-oxygen species in the reaction and lowers the pKa of the attached proton, thereby increasing the concentration of the attacking alkoxide nucleophile in an equilibrium process. In contrast, the role of the metal ion at binding site 2 is to catalyze the reaction by absorbing the negative charge that accumulates at the leaving 5'-oxygen in the transition state. We suggest structural reasons why the Mg(2+)-La3+ ion combination is particularly suited to demonstrating these different roles of the two-metal ions in the ribozyme cleavage reaction.

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Hepatitis C virus (HCV ) core (C) protein is thought to bind to viral RNA before it undergoes oligomerization leading to RNA encapsidation. Details of these events are so far unknown. The 5ʹ-terminal C protein coding sequence that includes an adenine (A)-rich tract is a part of an internal ribosome entry site(IRES). This nucleotide sequence but not the corresponding protein sequence is needed for proper initiation of translation of viral RNA by an IRES-dependent mechanism. In this study, we examined the importance of this sequence for the ability of the C protein to bind to viral RNA. Serially truncated C proteins with deletions from 10 up to 45 N-terminal amino acids were expressed in Escherichia coli, purified and tested for binding to viral RNA by a gel shift assay. The results showed that truncation of the C protein from its N-terminus by more than 10 amino acids abolished almost completely its expression in E. coli. The latter could be restored by adding a tag to the N-terminus of the protein. The tagged proteins truncated by 15 or more amino acids showed an anomalous migration in SDS-PAGE. Truncation by more than 20 amino acids resulted in a complete loss of ability of tagged C protein to bind to viral RNA. These results provide clues to the early events in the C protein - RNA interactions leading to C protein oligomerization, RNA encapsidation and virion assembly.

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The film adaptation of "Rosencrantz and Guildenstern Are Dead"'s constant reallocation of actor and audience roles (or subject and object positions) means that the film’s viewers are as deeply implicated in considering issues of identity, agency and determination as Rosencrantz and Guildenstern are. Tellingly, one of The Player’s outbursts reveals the philosophical connections between observing and being observed in ways that are true of the theatre, but which also transcend it: ‘You don’t understand the humiliation of it. To be tricked out of the single assumption that makes our existence bearable; that somebody is watching.’ In this statement is one of the film’s main concerns; that is, the relationship between knowing the self, knowing others, and being known by others.

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Objective: To explore psychosocial issues perceived to impact the mental health and well-being of resident (non-fly-in fly-out) mine workers at a local mine in regional Queensland. Design: A descriptive qualitative study using semistructured interviews. Setting: The research was conducted on-site at an opencut coal mine in regional Queensland. Participants: Ten miners (nine men) currently employed in workshop, production or supervisory roles. Main outcome measures: Self-reported issues affecting psychological well-being. Results: Participants’ occupation and the surrounding context appeared to have both positive and negative influences on their well-being. Overall findings could be grouped into four key themes: (i) the importance of relationships; (ii) the impact of lifestyle; (iii) work characteristics; and (iv) mental health attitudes. While not without strains on mental health, in general, participants reported that their current situation was superior to their previous mining jobs. This was attributed to close relationships among locally recruited workers, respect for management practices and rosters that allowed adequate sleep recovery and family time between shifts. Conclusions: This study is the first to examine mental health and well being in non-fly-in fly-out mining populations. It suggests that while some issues appear inherent in the mining occupation, personal and organisational support can help workers have a more positive workplace experience. Further work looking at more extensive comparisons over various mining contexts will greatly assist in the development of programs and support structures for rural and regional mine workers.

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Background: HIV-1 Gag virus like particles (VLPs) used as candidate vaccines are regarded as inert particles as they contain no replicative nucleic acid, although they do encapsidate cellular RNAs. During HIV-1 Gag VLP production in baculovirus-based expression systems, VLPs incorporate the baculovirus Gp64 envelope glycoprotein, which facilitates their entry into mammalian cells. This suggests that HIV-1 Gag VLPs produced using this system facilitate uptake and subsequent expression of encapsidated RNA in mammalian cells - an unfavourable characteristic for a vaccine. Methods. HIV-1 Gag VLPs encapsidating reporter chloramphenicol acetyl transferase (CAT) RNA, were made in insect cells using the baculovirus expression system. The presence of Gp64 on the VLPs was verified by western blotting and RT-PCR used to detect and quantitate encapsidated CAT RNA. VLP samples were heated to inactivate CAT RNA. Unheated and heated VLPs incubated with selected mammalian cell lines and cell lysates tested for the presence of CAT protein by ELISA. Mice were inoculated with heated and unheated VLPs using a DNA prime VLP boost regimen. Results: HIV-1 Gag VLPs produced had significantly high levels of Gp64 (∼1650 Gp64 molecules/VLP) on their surfaces. The amount of encapsidated CAT RNA/g Gag VLPs ranged between 0.1 to 7 ng. CAT protein was detected in 3 of the 4 mammalian cell lines incubated with VLPs. Incubation with heated VLPs resulted in BHK-21 and HeLa cell lysates showing reduced CAT protein levels compared with unheated VLPs and HEK-293 cells. Mice inoculated with a DNA prime VLP boost regimen developed Gag CD8 and CD4 T cell responses to GagCAT VLPs which also boosted a primary DNA response. Heating VLPs did not abrogate these immune responses but enhanced the Gag CD4 T cell responses by two-fold. Conclusions: Baculovirus-produced HIV-1 Gag VLPs encapsidating CAT RNA were taken up by selected mammalian cell lines. The presence of CAT protein indicates that encapsidated RNA was expressed in the mammalian cells. Heat-treatment of the VLPs altered the ability of protein to be expressed in some cell lines tested but did not affect the ability of the VLPs to stimulate an immune response when inoculated into mice. © 2011 Valley-Omar et al; licensee BioMed Central Ltd.

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The DNA damage response encompasses a complex series of signaling pathways that function to regulate and facilitate the repair of damaged DNA. Recent studies have shown that the repair of transcriptionally inactive chromatin, named heterochromatin, is dependent upon the phosphorylation of the co-repressor, Krüppel-associated box (KRAB) domain-associated protein (KAP-1), by the ataxia telangiectasia-mutated (ATM) kinase. Co-repressors, such as KAP-1, function to regulate the rigid structure of heterochromatin by recruiting histone-modifying enzymes, such HDAC1/2, SETDB1, and nucleosome-remodeling complexes such as CHD3. Here, we have characterized a phosphorylation site in the HP1-binding domain of KAP-1, Ser-473, which is phosphorylated by the cell cycle checkpoint kinase Chk2. Expression of a nonphosphorylatable S473A mutant conferred cellular sensitivity to DNA-damaging agents and led to defective repair of DNA double-strand breaks in heterochromatin. In addition, cells expressing S473A also displayed defective mobilization of the HP1-β chromodomain protein. The DNA repair defect observed in cells expressing S473A was alleviated by depletion of HP1-β, suggesting that phosphorylation of KAP-1 on Ser-473 promotes the mobilization of HP1-β from heterochromatin and subsequent DNA repair. These results suggest a novel mechanism of KAP-1-mediated chromatin restructuring via Chk2-regulated HP1-β exchange from heterochromatin, promoting DNA repair.

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