141 research outputs found

    Dissecting the mechanisms of transport of herpes simplex virus between Langerhans Cells & dendritic cells in epidermis and dermis following infection of human genital mucosa and skin

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    Herpes Simplex Virus (HSV) is a sexually transmitted infection (STI) that the World Health Organisation (WHO) has deemed a priority for a vaccine. CD8 and CD4T cells are important in the control and clearance of HSV, however no known vaccine has been able to stimulate CD8T cells. The dermal dendritic cells (dDCs) are suspected to play a role. Previously the host lab has shown in human tissue that HSV-1 infection of Langerhans cells (LCs) caused apoptosis and migration of LCs to the dermis, where they were phagocytosed by dDCs (termed HSV viral relay). Very little is known about the mechanisms of this relay. The host lab has also identified a second resident epidermal immune cell, Epi-cDC2s, which are infectable by HSV. This thesis aims to unravel the mechanisms involved in the relay. RNA-seq and cell surface phenotyping on human dDCs subsets showed that was differential chemokine receptor expression. Bead-based immunoassays were used to determine the chemokines produced by HSV-1 infected LCs and Epi-cDC2s,and showed HSV infected LCs produced increased CXCR3 ligands, while HSV infected Epi-cDC2s produced increased CCR5 ligands. The importance of these chemokine axes was investigated using chemotaxis assays. An cyclic immunofluorescent microscopy panel was then developed to investigate whether this migration could be seen in situ in HSV infected foreskin explants. Underneath epidermal foci of infection, there was migration of both cDC1s and cDC2s towards the basement membrane. Under foci of infection there was a greater proportion of cDC2s clustering with LCs. The uptake of HSV infected epidermal cells by the dDC subsets was examined using imaging cytometry. Preliminary results suggest that there were no significant differences between the ability of dDCs to phagocytose HSV infected epidermal cells. Understanding the mechanisms and the role of each dDC subset in the HSV viral relay will determine which dDC subsets are crucial for CD8 and CD4 T cell stimulation

    A Novel Anthropomorphic Flow Phantom for the Quantitative Evaluation of Prostate DCE-MRI Acquisition Techniques

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    A novel anthropomorphic flow phantom device has been developed which can be used for quantitatively assessing the ability of MRI scanners to accurately measure signal / concentration time-intensity curves (CTCs) associated with dynamic contrast-enhanced (DCE) MRI. Modelling of the complex pharmacokinetics of contrast agents as they perfuse through the tumour capillary network has shown great promise for cancer diagnosis and therapy monitoring. However, clinical adoption has been hindered by methodological problems, resulting in a lack of consensus regarding the most appropriate acquisition and modelling methodology to use and a consequent wide discrepancy in published data. A heretofore overlooked source of such discrepancy may arise from measurement errors of tumour CTCs deriving from the imaging pulse sequence itself, while the effects on the fidelity of CTC measurement of using rapidly-accelerated sequences such as parallel imaging and compressed sensing remain unknown. The present work aimed to investigate these features by developing a test device in which „ground truth‟ CTCs were generated and presented to the MRI scanner for measurement, thereby allowing for an assessment of the DCE-MRI protocol to accurately measure this curve-shape. The device comprised of a 4-pump flow system wherein CTCs derived from prior patient prostate data were produced in measurement chambers placed within the imaged volume. The ground truth was determined as the mean of repeat measurements using an MRI-independent, custom-built optical imaging system. In DCE-MRI experiments, significant discrepancies between the ground truth and measured CTCs were found for both tumorous and healthy tissue-mimicking curve shapes. Pharmacokinetic modelling revealed errors in measured Ktrans, ve and kep values of up to 42%, 31%, and 50% respectively, following a simple variation of the parallel imaging factor and number of signal averages in the acquisition protocol The device allows for the quantitative assessment and standardisation of DCE-MRI protocols (both existing and emerging)

    Human Powered Vehicle Trainer

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    This Final Design Review (FDR) document describes the final design and completed prototype of a Mechanical Engineering senior project team at California Polytechnic State University, San Luis Obispo. The project goal is to create an adjustable human powered vehicle training bike for George Leone that allows a rider to gain confidence with the unique reclined bike geometry ahead of the World Human Powered Speed Challenge at Battle Mountain, Nevada. This document outlines the customer’s needs and technical research performed which together determine the project’s scope and engineering specifications. Next, we present the initial idea generation process and its results, along with the top mechanism designs, the decision matrices used to evaluate them, and the final design concepts. Each finalized subsystem design is then presented, including all required materials and components. Following this, the manufacturing section details the final project budget and the build processes followed for each subsystem and the overall bike. To ensure the bike met its design specifications, tests were performed with the completed prototype and their results are presented in the testing section. Additionally, we illustrate the overall project management plan and Gantt chart, and scheduling lessons learned from our project. Finally, the conclusion presents our final thoughts on the project – including what went well and what we would do differently – along with recommendations for our sponsor on how to improve and utilize the trainer in the future

    Local GABA concentration is related to network-level resting functional connectivity

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    Anatomically plausible networks of functionally inter-connected regions have been reliably demonstrated at rest, although the neurochemical basis of these ‘resting state networks’ is not well understood. In this study, we combined magnetic resonance spectroscopy (MRS) and resting state fMRI and demonstrated an inverse relationship between levels of the inhibitory neurotransmitter GABA within the primary motor cortex (M1) and the strength of functional connectivity across the resting motor network. This relationship was both neurochemically and anatomically specific. We then went on to show that anodal transcranial direct current stimulation (tDCS), an intervention previously shown to decrease GABA levels within M1, increased resting motor network connectivity. We therefore suggest that network-level functional connectivity within the motor system is related to the degree of inhibition in M1, a major node within the motor network, a finding in line with converging evidence from both simulation and empirical studies

    Measuring cosmic density of neutral hydrogen via stacking the DINGO-VLA data

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    We use the 21-cm emission-line data from the Deep Investigation of Neutral Gas Origin-Very Large Array (DINGO-VLA) project to study the atomic hydrogen gas H I of the Universe at redshifts z \u3c 0.1. Results are obtained using a stacking analysis, combining the H I signals from 3622 galaxies extracted from 267 VLA pointings in the G09 field of the Galaxy and Mass Assembly Survey (GAMA). Rather than using a traditional one-dimensional spectral stacking method, a three-dimensional cubelet stacking method is used to enable deconvolution and the accurate recovery of average galaxy fluxes from this high-resolution interferometric data set. By probing down to galactic scales, this experiment also overcomes confusion corrections that have been necessary to include in previous single-dish studies. After stacking and deconvolution, we obtain a 30σ H I mass measurement from the stacked spectrum, indicating an average H I mass of MHI=(1.67±0.18)×109 M⊙ role= presentation style= box-sizing: border-box; margin: 0px; padding: 0px; border: 0px; font-variant: inherit; font-stretch: inherit; line-height: normal; font-family: inherit; vertical-align: baseline; display: inline; word-spacing: normal; overflow-wrap: normal; white-space: nowrap; float: none; direction: ltr; max-width: none; max-height: none; min-width: 0px; min-height: 0px; position: relative; \u3eMHI=(1.67±0.18)×109 M⊙MHI=(1.67±0.18)×109 M⊙⁠. The corresponding cosmic density of neutral atomic hydrogen is ΩHI=(0.38±0.04)×10−3 role= presentation style= box-sizing: border-box; margin: 0px; padding: 0px; border: 0px; font-variant: inherit; font-stretch: inherit; line-height: normal; font-family: inherit; vertical-align: baseline; display: inline; word-spacing: normal; overflow-wrap: normal; white-space: nowrap; float: none; direction: ltr; max-width: none; max-height: none; min-width: 0px; min-height: 0px; position: relative; \u3eΩHI=(0.38±0.04)×10−3ΩHI=(0.38±0.04)×10−3 at redshift of z = 0.051. These values are in good agreement with earlier results, implying there is no significant evolution of ΩHI role= presentation style= box-sizing: border-box; margin: 0px; padding: 0px; border: 0px; font-variant: inherit; font-stretch: inherit; line-height: normal; font-family: inherit; vertical-align: baseline; display: inline; word-spacing: normal; overflow-wrap: normal; white-space: nowrap; float: none; direction: ltr; max-width: none; max-height: none; min-width: 0px; min-height: 0px; position: relative; \u3eΩHIΩHI at lower redshifts

    Mechanisms of Immune Control of Mucosal HSV Infection: A Guide to Rational Vaccine Design

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    Herpes Simplex Virus (HSV) is a highly prevalent sexually transmitted infection that aside from causing cold sores and genital lesions, causes complications in the immunocompromised and has facilitated a large proportion of HIV acquisition globally. Despite decades of research, there is no prophylactic HSV vaccine ready for use in humans, leaving many questioning whether a prophylactic vaccine is an achievable goal. A previous HSV vaccine trial did have partial success in decreasing acquisition of HSV2–promising evidence that vaccines can prevent acquisition. However, there is still an incomplete understanding of the immune response pathways elicited by HSV after initial mucosal infection and how best to replicate these responses with a vaccine, such that acquisition and colonization of the dorsal root ganglia could be prevented. Another factor to consider in the rational design of an HSV vaccine is adjuvant choice. Understanding the immune responses elicited by different adjuvants and whether lasting humoral and cell-mediated responses are induced is important, especially when studies of past trial vaccines found that a sufficiently protective cell-mediated response was lacking. In this review, we discuss what is known of the immune control involved in initial herpes lesions and reactivation, including the importance of CD4 and CD8 T cells, and the interplay between innate and adaptive immunity in response to primary infection, specifically focusing on the viral relay involved. Additionally, a summary of previous and current vaccine trials, including the components used, immune responses elicited and the feasibility of prophylactic vaccines looking forward, will also be discussed

    Insights into the Role of a Cardiomyopathy-Causing Genetic Variant in ACTN2

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    Pathogenic variants in ACTN2, coding for alpha-actinin 2, are known to be rare causes of Hyper-trophic Cardiomyopathy. However, little is known about the underlying disease mechanisms. Adult heterozygous mice carrying the Actn2 p.Met228Thr variant were phenotyped by echocar-diography. For homozygous mice, viable E15.5 embryonic hearts were analysed by High Reso-lution Episcopic Microscopy and wholemount staining, complemented by unbiased proteomics, qPCR and Western blotting. Heterozygous Actn2 p.Met228Thr mice have no overt phenotype. Only mature males show molecular parameters indicative of cardiomyopathy. By contrast, the variant is embryonically lethal in the homozygous setting and E15.5 hearts show multiple morphological abnormalities. Molecular analyses, including unbiased proteomics, identified quantitative abnormalities in sarcomeric parameters, cell cycle defects and mitochondrial dys-function. The mutant alpha-actinin protein is found to be destabilised, associated with increased activity of the ubiquitin-proteosomal system. This missense variant in alpha-actinin renders the protein less stable. In response, the ubiquitin-proteosomal system is activated; a mechanism which has been implicated in cardiomyopathies previously. In parallel, lack of functional al-pha-actinin is thought to cause energetic defects through mitochondrial dysfunction. This seems, together with cell cycle defects, the likely cause of death of the embryos. The defects also have wide-ranging morphological consequences

    Phenotypic differentiation among native, expansive and introduced populations influences invasion success

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    Aim: Humans influence species distributions by modifying the environment and by dispersing species beyond their natural ranges. Populations of species that have established in disjunct regions of the world may exhibit trait differentiation from native populations due to founder effects and adaptations to selection pressures in each distributional region. We compared multiple native, expansive and introduced populations of a single species across the world, considering the influence of environmental stressors and transgenerational effects. Location: United States Gulf and Atlantic coasts, United States interior, European Atlantic and Mediterranean coasts, east coast of Australia. Taxon: Baccharis halimifolia L. (eastern baccharis). Methods: We monitored seed germination, seedling emergence, survival and early growth in a common garden experiment, conducted with over 18,200 seeds from 80 populations. We also evaluated the influence of environmental stress and maternal traits on progeny performance. Results: Introduced European Atlantic populations had faster germination and early growth than native populations. However, this was not the case for the more recently naturalized European Mediterranean populations. Introduced Australian populations grew faster than native populations in non-saline environments but had lower survival in saline conditions commonly encountered in the native range. Similarly, expansive inland US populations germinated faster than coastal native populations in non-saline environments but grew and germinated more slowly in saline environments. Maternal inflorescence and plant size were positively related with seed germination and seedling survival, whereas flower abundance was positively correlated with seedling early growth and survival. However, maternal traits explained a much lower fraction of the total variation in early demographic stages of B. halimifolia than did distributional range. Main conclusions: Phenotypic differentiation could allow B. halimifolia to adapt to different biotic and abiotic selection pressures found in each distributional range, potentially contributing to its success in introduced and expansive ranges
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