3,311 research outputs found

    Practical Applications and Future Directions of Genetic Code Expansion: Validation of Novel Akt1 Substrates and the Design of a Synthetic Auxotroph Strain of B. subtilis

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    In Chapter 1, site-specifically phosphorylated variants of the oncogene Akt1 were made in Escherichia coli using the orthogonal translation system that enable genetic code expansion with phosphoserine. The differentially phosphorylated variants of Akt1 were used to validate newly predicted Akt1 substrates. The predicted target sites of the peptide substrates were synthesized and subjected to in vitro kinase assays to quantify the activity of each Akt1 phosphorylated variant towards the predicted peptide. A previously uncharacterized kinase-substrate interaction between Akt1 and a peptide derived from RAB11 Family Interacting Protein 2 (RAB11FIP2) was validated in vitro. Chapter 2 describes the preliminary development of a novel orthogonal translation system for Bacillus subtilis. The work presented outlines the design process: from selection of the components to the generation of an all-in-one plasmid containing the orthogonal translation system. The work demonstrates stable integration of the orthogonal translation system into the B. subtilis genome

    The Heart of the Matter: The Core and the Task of Core Revision

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    Gene expression: degrade to derepress

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    Chromatin immunoprecipitation and sequencing (ChIP‐seq) provides a static snap‐shot of DNA‐associated proteins which fails to reflect the dynamics of the DNA‐bound proteome. Now, Catic and co‐workers combine ubiquitin ChIP‐seq and proteasome inhibitors to map sites of DNA‐associated protein degradation on a genome‐wide scale. They identify an ubiquitin ligase which targets a transcriptional repressor for destruction by the proteasome, thus activating transcription of specific genes. These findings reveal that the ubiquitin proteasome system actively regulates transcription

    Novel stacked folded cores for blast-resistant sandwich beams

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    Recent research has established the effectiveness of sandwich structures with metallic cellular cores for blast mitigation. The choice of core architecture can enhance sandwich performance, dissipating energy through plastic core compression and exploiting fluid-structure interaction effects to reduce the momentum imparted to the structure by the blast. In this paper we describe the first analysis of a novel sandwich core concept for blast mitigation: the Stacked Folded Core. The core consists of an alternating stacked sequence of folded sheets in the Miura (double-corrugated) pattern, with the stack oriented such that the folding kinematics define the out-of plane compressive strength of the core. It offers a number of distinct characteristics compared to existing cellular cores. (i) The kinematics of collapse of the core by a distinctive folding mechanism give it unique mechanical properties, including strong anisotropy. (ii) The fold pattern and stacking arrangement is extremely versatile, offering exceptional freedom to tailor the mechanical properties of the core. This includes freedom to grade the core properties through progressive changes in the fold pattern. (iii) Continuous manufacturing processes have been established for the Miura folded sheets which make up the core. The design is therefore potentially more straightforward and economical to manufacture than other metallic cellular materials. In this first investigation of the Stacked Folded Core, finite element analysis is used to investigate its characteristics under both quasi-static and dynamic loading. A dynamic analysis of an impulsively loaded sandwich beam with a stacked folded core reveals the versatility of the concept for blast mitigation. By altering the fold pattern alone, the durations of key phases of the dynamic sandwich response (core compression, beam bending) can be controlled. By altering both fold pattern and sheet thickness in the core, the same is achieved without altering the density of the core or the mass distribution of the sandwich beam.This is the author's accepted manuscript. The final version is available from Elsevier at: http://www.sciencedirect.com/science/article/pii/S0020768314003035

    Host Presidents\u27 Address: A Discussion on Ways Catholic Higher Education Institutions Can Assist Catholic Elementary and Secondary Schools

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    As part of the third Catholic Higher Education Collaborative Conference (CHEC), an event cosponsored by Boston College and Fordham University, the host university presidents, Rev. William P. Leahy, S.J., and Rev. Joseph M. McShane, S.J., addressed conference attendees and discussed ways higher education institutions can assist Catholic elementary and secondary schools. This article contains a transcript of their remarks. Fr. Leahy, the 25th president of Boston College, has a keen interest in Catholic schools, understanding their importance for the nation and for handing on the Catholic tradition to the next generation. Through his efforts, the Center for Catholic Education at Boston College, now the Roche Center for Catholic Education, was established. Fr. McShane, the 32nd president of Fordham University, shares a strong commitment to Catholic education. He has become one of the most powerful voices in New York City speaking on behalf of the need for Catholic schools. The work of Fordham’s Graduate School of Education and Center for Catholic School Leadership with Catholic schools throughout the metropolitan region has received support from Fr. McShane

    Identification of major factors influencing ELISpot-based monitoring of cellular responses to antigens from mycobacterium tuberculosis

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    A number of different interferon-c ELISpot protocols are in use in laboratories studying antigen-specific immune responses. It is therefore unclear how results from different assays compare, and what factors most significantly influence assay outcome. One such difference is that some laboratories use a short in vitro stimulation period of cells before they are transferred to the ELISpot plate; this is commonly done in the case of frozen cells, in order to enhance assay sensitivity. Other differences that may be significant include antibody coating of plates, the use of media with or without serum, the serum source and the number of cells added to the wells. The aim of this paper was to identify which components of the different ELISpot protocols influenced assay sensitivity and inter-laboratory variation. Four laboratories provided protocols for quantifying numbers of interferon-c spot forming cells in human peripheral blood mononuclear cells stimulated with Mycobacterium tuberculosis derived antigens. The differences in the protocols were compared directly. We found that several sources of variation in assay protocols can be eliminated, for example by avoiding serum supplementation and using AIM-V serum free medium. In addition, the number of cells added to ELISpot wells should also be standardised. Importantly, delays in peripheral blood mononuclear cell processing before stimulation had a marked effect on the number of detectable spot forming cells; processing delay thus should be minimised as well as standardised. Finally, a pre-stimulation culture period improved the sensitivity of the assay, however this effect may be both antigen and donor dependent. In conclusion, small differences in ELISpot protocols in routine use can affect the results obtained and care should be given to conditions selected for use in a given study. A pre-stimulation step may improve the sensitivity of the assay, particularly when cells have been previously frozen

    Mapping Learning and Game Mechanics for Serious Games Analysis in Engineering Education

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    In a world where students are increasing digitally tethered to powerful, ‘always on’ mobile devices, new models of engagement and approaches to teaching and learning are required from educators. Serious Games (SG) have proved to have instructional potential but there is still a lack of methodologies and tools not only for their design but also to support game analysis and assessment. This paper explores the use of SG to increase student engagement and retention. The development phase of the Circuit Warz game is presented to demonstrate how electronic engineering education can be radically reimagined to create immersive, highly engaging learning experiences that are problem-centered and pedagogically sound. The Learning Mechanics–Game Mechanics (LM-GM) framework for SG game analysis is introduced and its practical use in an educational game design scenario is shown as a case study

    "This town can't be that harmful": risk perception of lead exposure

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    Introduction: Much of the focus of lead risk health campaigns has been on vulnerable populations such as children and pregnant women, thus not communicating the risk of exposure for other adults. This becomes a particular issue for adults who are long-term residents of communities near lead mines as they can be at an increased risk of exposure to lead within their environment. As such, this study investigated the perceived risk of exposure to lead in residents of a lead mining community. Methods: Semi-structured interviews were held with 20 (3 male, 17 female) residents from a community in close proximity to a lead mine and were recruited through the community media and local organizations. Common themes were identified through an interpretative phenomenological analytical framework providing an in depth examination of the lived experiences of participants. Results: Majority of the participants did not perceive a health risk from exposure to lead. Those who reported a specific concern surrounding their exposure to lead had lived within the community for less than five years. However, it was commonly noted that the behaviors to control residents' exposure to lead were easily performed and low cost. Conclusions: These results suggest that residents of a community chronically exposed to lead seem to become complacent about their risk for poor health outcomes the longer they live within the community. These findings have implications for the communication of the risk of exposure to lead for adults who are chronically exposed to lead
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