Glutamate release mechanisms from megakaryocytes

Cardiovascular disease (CVD) is one of the main causes of death in western society. Platelet activation, thrombus formation and plaque rupture are all central events in the pathogenesis of acute coronary syndromes, therefore therapies targeted at controlling platelet numbers and aggregation are likely to be beneficial in the treatment of CVD. Megakaryocytes (MKs) which are the precursors to platelets are an ideal target for these therapies, however the intrinsic factors that regulate the production and shedding of platelet precursors are poorly understood. Recent studies identified that MKs express functional NMDA-type glutamate receptors similar to those found in the CNS and that antagonism of these receptors prevents normal MK differentiation and platelet function. This thesis investigates glutamate signalling within MKs further, focusing on the glutamate release capability of MK cells and the mechanisms involved. Using molecular and cellular techniques it was demonstrated that MK cells expressed numerous regulatory proteins required for vesicular glutamate release, including core SNARE proteins, VAMP, SNAP-23 and syntaxin; specific glutamate-loading vesicle proteins, VGLUTI and VGLUT2; and glutamate transporters, EAATI and EAAT2. Active vesicle recycling was observed in MK cells using a fluorescent reporter and an enzyme-linked fluorimetric assay confirmed that MK cells constitutively released glutamate and that glutamate release levels increased significantly following MK differentiation. Transient transfection of the human cell line MEG-Ol with tetanus toxin, which disables vesicle recycling, induced a 30% decrease (P<O.OOI) in released glutamate compared to empty vector controls. In contrast, over-expressing VGLUTI caused a 41% increase (P<O.OO1) in glutamate release activity of MEG-O1 cells compared to controls. These data demonstrate that MK cells regulate glutamate exocytosis through specific vesicular proteins, indicating that glutamate signalling may be a potential target for CVD therapies. Also the observations that MKs both release and recycle glutamate indicates an important role for glutamate signalling from these cells in autocrine and paracrine interactions within the bone marrow microenvironment

Soft X-ray Photochemistry at the L₂,₃-edges in K₃[Fe(CN)₆], [Co(acac)₃] and [Cp₂Fe][BF₄]

Soft X-ray photoreduction has been observed in K3[Fe(CN)6], [Co(acac)3] and [Cp2Fe][BF4]

Agreement between mothers and children with malocclusion in rating children's oral health-related quality of life

Introduction: The aim of this study was to compare the assessment of oral health-related quality of life (OH-QoL) between children with malocclusion and their mothers, by using responses to the child perceptions questionnaire and the parental-caregivers perceptions questionnaire. Methods: The study was conducted in 90 children, aged 11 to 14 years, with a malocclusion grade of 4 or 5 according to the index of orthodontic treatement need dental health component. The children and their mothers completed the questionnaires independently. Results: The mean ratings were similar for total scores (children, 20.4; mothers, 20.1), oral symptoms (children, 5.2; mothers, 4.7), and social well-being (children, 4.3; mothers, 4.8). However, the mothers group had a lower mean score for functional limitations (children, 5.3; mothers, 3.6) and a higher mean score for emotional well-being (children, 5.6; mothers, 7.1). The correlations between children's and mothers' responses ranged from rs = 0.545 for total score and emotional well-being to rs = 0.357 for functional limitations. There were good correlations between their responses to global (rs = 0.466) and life overall (rs = 0.427) questions, but poor correlations between the 2 questions, suggesting that these concepts were considered differently. Conclusions: Maternal opinions were similar to those of their children for the overall impact on OH-QoL of malocclusion, but mothers were more dissatisfied with the appearance of their children's teeth and overestimated the emotional impact of malocclusion. It would be useful to develop a specific measure to assess OH-QoL in children with malocclusion. (Am J Orthod Dentofacial Orthop 2010;137:631-8

Gettysburg College Sustainability Proposal

In the fall of 2011, the Environmental Studies capstone class led by Professor Rutherford Platt was asked to write Gettysburg College’s first Sustainability Plan. The goal of the plan was to develop specific sustainable practices for the campus that were related to the three pillars of sustainability: economic, social, and environmental, and how integrating diligent sustainable practices into each of these respected pillars will result in a more conscious campus, community, and future. In 2010, Gettysburg College turned to the Sustainability Tracking Assessment and Rating System (STARS) to quantify the institution’s sustainability efforts, providing a self-check mechanism to encourage sustainability applications to all aspects of the College. The American College and University Presidents’ Climate Commitment was signed in 2007 by former Gettysburg College President Katherine Haley Will, declaring that Gettysburg College would become carbon neutral by 2032. Gettysburg College has made large strides in the search for sustainability, and aims to continue its dedication to furthering sustainable practice. The following plan outlines the six priority areas identified by the Capstone class: progress of the American College and University Presidents’ Climate Commitment, Dining Services, campus green space, community outreach, integration of sustainability into the Gettysburg College Curriculum, and the Sustainability Advisory Committee. The first priority area identified was monitoring and upholding the American College and University Presidents’ Climate Commitment (ACUPCC). Though creating new sustainability initiatives on campus is the driving force towards an increasingly sustainable college and community, it is imperative that these goals be carried out in full to maximize beneficial returns. In order to reach carbon neutrality, Gettysburg College hopes to increase energy efficiency in buildings, incorporate renewable energy sources on campus, and mitigate remaining emissions through the purchase of carbon offsets. To further the College’s progress, it is proposed that Gettysburg College continue its energy-efficient appliance purchasing policy, as well as create a policy to offset all greenhouse gas emissions generated by air travel for students study abroad. As stated by the ACUPCC, a Sustainability Committee should take responsibility for the updates and progress reports required to meet the goal of carbon neutrality. The second priority area identified was sustainability in Dining Services. Gettysburg College is home to 2,600 students, all of whom require three full meals a day. Dining Services accounts for a large fraction of Gettysburg College’s sustainability efforts, already implementing sustainability through composting, buying local produce, and using biodegradable products. The proposed on-campus sales cuts of non-reusable to-go items, a change in campus mentality on food waste, and improved composting practices will translate to an increasingly sustainable campus, as well as a well-fed campus body. The third priority was maintaining green space on campus. Ranked as the 23rd most beautiful campus in the United States by The Best Colleges, Gettysburg College utilizes campus green space to create an atmosphere that is conducive to activity as well as tranquility. The plan proposes that Gettysburg College and its grounds facilities continue their exceptional efforts, focusing on increasing the use of the student garden, creating a new rain garden or social area on campus, and converting unnecessary parking lots into green space. As these additions are completed, they must be introduced to the student body and faculty alike to assure these areas are known and utilized. The fourth priority was utilizing community outreach to spread awareness of sustainability initiatives on and off campus. To connect the sustainability-geared changes proposed in this plan, community outreach at Gettysburg College is assessed to estimate how well these initiatives are communicated and promoted to both potential and enrolled students, faculty, and other concerned parties. To evaluate the efficiency of communication at Gettysburg College, a quantitative assessment is presented to measure the ease of finding the sustainability webpage, the quality of sustainability-related topics available on the webpage, and quality of webpage design. The webpage is in need of improved text to image ratios, locations of sustainability topics, and data displays. Despite not having a link to the sustainability webpage on the Gettysburg College homepage, sustainability events should be covered and presented on the rotational news feed found on the homepage to maximize outreach to interested parties or simply to add to the definition of Gettysburg College. The fifth priority was integrating sustainability into the Curriculum to build a culture on campus that values academic rigor, supports students as they cultivate intellectual and civic passions, and promotes the development of healthy social relationships and behaviors. The proposed Sustainability Committee on Sustainability in the Curriculum (SCC) will hold sustainability workshops for faculty with the aim to instill sustainability into all academic disciplines, providing all Gettysburg graduates with a means to approach their professional careers in a fashion that is conscious of sustainability. The sixth and last priority was the Sustainability Advisory Committee. Established in 2007, the Sustainability Advisory Committee is currently under review, but it is recommended that the committee restructure itself in accordance with the new Sustainability Committee Bylaws. These bylaws aim to define the purposes, membership, governance, and involvement with the college. With a clearly defined set of goals and methodology, the Sustainability Advisory Committee will be able to improve the solidarity of the sustainability movement on campus as a whole. By following the propositions laid out in the Gettysburg College Sustainability Plan, the student body, faculty, and community alike will become a part of a multi-faceted progression toward a more sustainable future

Pathomx:an interactive workflow-based tool for the analysis of metabolomic data

BACKGROUND: Metabolomics is a systems approach to the analysis of cellular processes through small-molecule metabolite profiling. Standardisation of sample handling and acquisition approaches has contributed to reproducibility. However, the development of robust methods for the analysis of metabolomic data is a work-in-progress. The tools that do exist are often not well integrated, requiring manual data handling and custom scripting on a case-by-case basis. Furthermore, existing tools often require experience with programming environments such as MATLAB® or R to use, limiting accessibility. Here we present Pathomx, a workflow-based tool for the processing, analysis and visualisation of metabolomic and associated data in an intuitive and extensible environment. RESULTS: The core application provides a workflow editor, IPython kernel and a HumanCyc™-derived database of metabolites, proteins and genes. Toolkits provide reusable tools that may be linked together to create complex workflows. Pathomx is released with a base set of plugins for the import, processing and visualisation of data. The IPython backend provides integration with existing platforms including MATLAB® and R, allowing data to be seamlessly transferred. Pathomx is supplied with a series of demonstration workflows and datasets. To demonstrate the use of the software we here present an analysis of 1D and 2D (1)H NMR metabolomic data from a model system of mammalian cell growth under hypoxic conditions. CONCLUSIONS: Pathomx is a useful addition to the analysis toolbox. The intuitive interface lowers the barrier to entry for non-experts, while scriptable tools and integration with existing tools supports complex analysis. We welcome contributions from the community. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12859-014-0396-9) contains supplementary material, which is available to authorized users

Metagenomic characterisation of the viral community of lough neagh, the largest freshwater lake in Ireland

Lough Neagh is the largest and the most economically important lake in Ireland. It is also one of the most nutrient rich amongst the world's major lakes. In this study, 16S rRNA analysis of total metagenomic DNA from the water column of Lough Neagh has revealed a high proportion of Cyanobacteria and low levels of Actinobacteria, Acidobacteria, Chloroflexi, and Firmicutes. The planktonic virome of Lough Neagh has been sequenced and 2,298,791 2×300 bp Illumina reads analysed. Comparison with previously characterised lakes demonstrates that the Lough Neagh viral community has the highest level of sequence diversity. Only about 15% of reads had homologs in the RefSeq database and tailed bacteriophages (Caudovirales) were identified as a major grouping. Within the Caudovirales, the Podoviridae and Siphoviridae were the two most dominant families (34.3% and 32.8% of the reads with sequence homology to the RefSeq database), while ssDNA bacteriophages constituted less than 1% of the virome. Putative cyanophages were found to be abundant. 66,450 viral contigs were assembled with the largest one being 58,805 bp; its existence, and that of another 34,467 bp contig, in the water column was confirmed. Analysis of the contigs confirmed the high abundance of cyanophages in the water column