98 research outputs found

    Examination of Methylation Sites for Forensic Age Determination from Semen

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    Methylation Sensitive High-Resolution Melt (MS-HRM) is based on quantitating the melt curve from an experimental sample against a standard of known methylation levels. Whereas most applications of age prediction using methylation markers are based upon pyrosequencing or SNaPshot technologies, these analysis methods are both cost and instrumentation prohibitive. This study sought to use to the varied methylation status of the ELOVL2 and FHL2 alleles, both having known correlation with age (Hamano et. al.), in a labor and time efficient manner to develop an age prediction model. A non-linear regression and standard curve was compiled from the methylation status in a sample (n=7) of extracted semen samples and compared to chronological age. The methylation status of ELVOL2 and FHL2 from each sample was obtained, with the conclusion that no correlation in methylation percentage and biological age existed for this sample of individuals aged 20-33. The principal objective of this study, to expand the application of MS-HRM age prediction from blood to other body fluids, will need further testing using larger sample sizes and broader age ranges prior to application in forensic casework.https://scholarscompass.vcu.edu/uresposters/1281/thumbnail.jp

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    The Sodium Sialic Acid Symporter From Staphylococcus aureus Has Altered Substrate Specificity

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    Mammalian cell surfaces are decorated with complex glycoconjugates that terminate with negatively charged sialic acids. Commensal and pathogenic bacteria can use host-derived sialic acids for a competitive advantage, but require a functional sialic acid transporter to import the sugar into the cell. This work investigates the sodium sialic acid symporter (SiaT) from Staphylococcus aureus (SaSiaT). We demonstrate that SaSiaT rescues an Escherichia coli strain lacking its endogenous sialic acid transporter when grown on the sialic acids N-acetylneuraminic acid (Neu5Ac) or N-glycolylneuraminic acid (Neu5Gc). We then develop an expression, purification and detergent solubilization system for SaSiaT and demonstrate that the protein is largely monodisperse in solution with a stable monomeric oligomeric state. Binding studies reveal that SaSiaT has a higher affinity for Neu5Gc over Neu5Ac, which was unexpected and is not seen in another SiaT homolog. We develop a homology model and use comparative sequence analyses to identify substitutions in the substrate-binding site of SaSiaT that may explain the altered specificity. SaSiaT is shown to be electrogenic, and transport is dependent upon more than one Na+ ion for every sialic acid molecule. A functional sialic acid transporter is essential for the uptake and utilization of sialic acid in a range of pathogenic bacteria, and developing new inhibitors that target these transporters is a valid mechanism for inhibiting bacterial growth. By demonstrating a route to functional recombinant SaSiaT, and developing the in vivo and in vitro assay systems, our work underpins the design of inhibitors to this transporter

    The state of the Martian climate

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    60°N was +2.0°C, relative to the 1981–2010 average value (Fig. 5.1). This marks a new high for the record. The average annual surface air temperature (SAT) anomaly for 2016 for land stations north of starting in 1900, and is a significant increase over the previous highest value of +1.2°C, which was observed in 2007, 2011, and 2015. Average global annual temperatures also showed record values in 2015 and 2016. Currently, the Arctic is warming at more than twice the rate of lower latitudes

    Crystal, Solution and In silico Structural Studies of Dihydrodipicolinate Synthase from the Common Grapevine

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    Dihydrodipicolinate synthase (DHDPS) catalyzes the rate limiting step in lysine biosynthesis in bacteria and plants. The structure of DHDPS has been determined from several bacterial species and shown in most cases to form a homotetramer or dimer of dimers. However, only one plant DHDPS structure has been determined to date from the wild tobacco species, Nicotiana sylvestris (Blickling et al. (1997) J. Mol. Biol. 274, 608–621). Whilst N. sylvestris DHDPS also forms a homotetramer, the plant enzyme adopts a ‘back-to-back’ dimer of dimers compared to the ‘head-to-head’ architecture observed for bacterial DHDPS tetramers. This raises the question of whether the alternative quaternary architecture observed for N. sylvestris DHDPS is common to all plant DHDPS enzymes. Here, we describe the structure of DHDPS from the grapevine plant, Vitis vinifera, and show using analytical ultracentrifugation, small-angle X-ray scattering and X-ray crystallography that V. vinifera DHDPS forms a ‘back-to-back’ homotetramer, consistent with N. sylvestris DHDPS. This study is the first to demonstrate using both crystal and solution state measurements that DHDPS from the grapevine plant adopts an alternative tetrameric architecture to the bacterial form, which is important for optimizing protein dynamics as suggested by molecular dynamics simulations reported in this study

    Influence of psychological factors in food risk assessment - A review

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    Background: Typically, food-related risk assessments are carried out within a four step, technical framework, as detailed by the Codex Alimentarius Commission (World Health Organization/ Food and Agricultural Organization of the United Nations, 2015). However, the technical framework presumes a level of ‘objective risk’ and does not take into account that risk is complex and psychologically constructed, something which is rarely acknowledged within risk analysis as a whole. It is well documented that people's perceptions of risk are based on more than merely probability of occurrence, but reflect other non-technical psychological factors (e.g., risk origin, severity, controllability, familiarity). Moreover, the basis of these risk perceptions is largely similar for experts and non-experts. Scope and approach: In this review, we consider each stage of the risk assessment process from a psychological perspective, reviewing research on non-technical factors which could affect assessments of risk and subsequent risk management decisions, with a particular focus on food safety. Key Findings and Conclusions: We identify 12 factors from the psychological literature which could potentially influence how risks are assessed and characterised. Drawing on insights from this research, we propose a number of recommendations to standardise approaches in risk assessment. Acknowledging and working with the subjectivity of risk is key to ensuring the efficacy of the wider risk analysis process

    Prospective study of outcomes in Sporadic versus hereditary breast cancer (POSH): study protocol

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    Background: young women presenting with breast cancer are more likely to have a genetic predisposition to the disease than breast cancer patients in general. A genetic predisposition is known to increase the risk of new primary breast (and other) cancers. It is unclear from the literature whether genetic status should be taken into consideration when planning adjuvant treatment in a young woman presenting with a first primary breast cancer. The primary aim of the POSH study is to establish whether genetic status influences the prognosis of primary breast cancer independently of known prognostic factors. Methods/design: the study is a prospective cohort study recruiting 3,000 women aged 40 years or younger at breast cancer diagnosis; the recruiting period covers 1st June 2001 to 31st December 2007. Written informed consent is obtained at study entry. Family history and known epidemiological risk data are collected by questionnaire. Clinical information about diagnosis, treatment and clinical course is collected and blood is stored. Follow up data are collected annually after the first year. An additional recruitment category includes women aged 41 to 50 years who are found to be BRCA1 or BRCA2 gene carriers and were diagnosed with their first breast cancer during the study recruiting period. Discussion: power estimates were based on 10% of the cohort carrying a BRCA1 gene mutation. Preliminary BRCA1 and BRCA2 mutation analysis in a pilot set of study participants confirms we should have 97% power to detect a difference of 10% in event rates between gene carriers and sporadic young onset cases. Most of the recruited patients (>80%) receive an anthracycline containing adjuvant chemotherapy regimen making planned analyses more straightforwar

    Comparison of 6q25 Breast Cancer Hits from Asian and European Genome Wide Association Studies in the Breast Cancer Association Consortium (BCAC)

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