Orf Virus Genome Sequencing

Orf virus (ORFV) is an ancient Parapoxvirus that causes substantial economic loss worldwide to sheep and goat producers. This virus causes a disease known as Contagious Ecthyma or more commonly “Soremouth” because it most commonly presents itself on the lips and mouth of sheep and goats. Soremouth makes it difficult for animals to eat and drink, therefore leading to weight loss or failure to gain weight resulting in production losses. The primary victims of the disease are suckling lambs and kid goats. These animals have immature immune systems increasing their risk of infection and are at risk for dehydration and weight loss as the lesions in the mouth affect their ability to nurse. ORFV is a zoonotic virus that can transfer from sheep and goat to humans and other species. ORFV has immunomodulatory capabilities as the virus encodes a synthetic interleukin-10, an immune down-regulator. It also inhibits other immune activation pathways such as the Toll-like receptors. ORFV is an ideal target for vaccination because of its immunomodulatory features and because the infection is generally self-limiting. A vaccine is currently under development by Texas Vet Lab, Inc. and the genome of the virus used for the development of this vaccine is sequenced and examined here. The goal of this study was to allow comparisons between the current vaccine candidate and other sequenced ORFV genomes

QUANTITATIVE REAL-TIME PCR DETECTION OF PERKINSUS MARINUS AND HAPLOSPORIDIUM NELSONI IN TEXAS OYSTERS

Haplosporidium nelsoni (MSX) and Perkinsus marinus (Dermo) are protozoan parasites that are traditionally detected using time and labor intensive histological methods. Recently developed traditional PCR assays, specific for these parasites, were used to for initial screening of presence/absence in samples of the eastern oyster, Crassostrea virginica, collected from Galveston Bay, Aransas Bay, and Corpus Christi Bay, Texas. H. nelsoni (MSX) was not detected in any of the samples. P. marinus (dermo) was detected in oysters from all bays. Oysters that tested positive for P. marinus were further screened with quantitative PCR assays to enumerate the parasites. These data were directly compared to values obtained by Ray’s Fluid Thioglycollate histological method from the same sample. Though these tests have not been “ground-truthed” against the traditional histological methods it is the goal of this project to begin the process of comparing the two methods. There was strong agreement between the PCR and histological determination of P. marinus that is promising for eventual transition to PCR assays

Scapular body fractures—should we be fixing more of these?

The question of whether we should be fixing more scapular body fractures originates from the historical preference for nonoperative management of these fractures. Recently, there has been a renewed interest in operative management due to the recognition that scapular malunion can cause significant disability. While the treatment pendulum has shifted away from benign neglect, finding the right balance of surgical aggression remains controversial. In general, the majority of scapula fractures can successfully be treated nonoperatively with excellent functional results. However, numerous case studies exist demonstrating poor outcomes of scapular body or neck fractures with increased deformity. The literature suggests that a glenopolar angle (GPA) less than 20 degrees can lead to a significant decrease in shoulder function. Additionally, retrospective studies using lateral border offset (LBO) greater than >2 cm and angulation >45 degrees as a surgical indication demonstrate functional outcomes with near normal strength and range of motion and low complication rates. While numerous cut-offs for surgical indications have been recommended, all indications are considered relative and treatment should be individualized based on patient characteristics and goals

VASCULAR DYSFUNCTION IN THE Α-GALACTOSIDASE A-KNOCKOUT MOUSE IS AN ENDOTHELIAL CELL-, PLASMA MEMBRANE-BASED DEFECT

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/75176/1/j.1440-1681.2008.04984.x.pd

Substrate Reduction Augments the Efficacy of Enzyme Therapy in a Mouse Model of Fabry Disease

Fabry disease is an X-linked glycosphingolipid storage disorder caused by a deficiency in the activity of the lysosomal hydrolase α-galactosidase A (α-gal). This deficiency results in accumulation of the glycosphingolipid globotriaosylceramide (GL-3) in lysosomes. Endothelial cell storage of GL-3 frequently leads to kidney dysfunction, cardiac and cerebrovascular disease. The current treatment for Fabry disease is through infusions of recombinant α-gal (enzyme-replacement therapy; ERT). Although ERT can markedly reduce the lysosomal burden of GL-3 in endothelial cells, variability is seen in the clearance from several other cell types. This suggests that alternative and adjuvant therapies may be desirable. Use of glucosylceramide synthase inhibitors to abate the biosynthesis of glycosphingolipids (substrate reduction therapy, SRT) has been shown to be effective at reducing substrate levels in the related glycosphingolipidosis, Gaucher disease. Here, we show that such an inhibitor (eliglustat tartrate, Genz-112638) was effective at lowering GL-3 accumulation in a mouse model of Fabry disease. Relative efficacy of SRT and ERT at reducing GL-3 levels in Fabry mouse tissues differed with SRT being more effective in the kidney, and ERT more efficacious in the heart and liver. Combination therapy with ERT and SRT provided the most complete clearance of GL-3 from all the tissues. Furthermore, treatment normalized urine volume and uromodulin levels and significantly delayed the loss of a nociceptive response. The differential efficacies of SRT and ERT in the different tissues indicate that the combination approach is both additive and complementary suggesting the possibility of an improved therapeutic paradigm in the management of Fabry disease

Mannose 6-Phosphate Receptor and Sortilin Mediated Endocytosis of α-Galactosidase A in Kidney Endothelial Cells

Prominent vasculopathy in Fabry disease patients is caused by excessive intracellular accumulation of globotriaosylceramide (GL-3) throughout the vascular endothelial cells causing progressive cerebrovascular, cardiac and renal impairments. The vascular lesions lead to myocardial ischemia, atherogenesis, stroke, aneurysm, thrombosis, and nephropathy. Hence, injury to the endothelial cells in the kidney is a key mechanism in human glomerular disease and endothelial cell repair is an important therapeutic target. We investigated the mechanism of uptake of α-galactosidase A (α-Gal A) in renal endothelial cells, in order to clarify if the recombinant enzyme is targeted to the lysosomes via the universal mannose 6-phosphate receptor (M6PR) and possibly other receptors. Immunohistochemical localization of infused recombinant α-Gal A in a renal biopsy from a classic Fabry disease patient showed that recombinant protein localize in the endothelial cells of the kidney. Affinity purification studies using α-Gal A resins identified M6PR and sortilin as α-Gal A receptors in cultured glomerular endothelial cells. Immunohistochemical analyses of normal human kidney with anti-sortilin and anti-M6PR showed that sortilin and M6PR were expressed in the endothelium of smaller and larger vessels. Uptake studies in cultured glomerular endothelial cells of α-Gal A labeled with fluorescence and 125I showed by inhibition with RAP and M6P that sortilin and M6PR mediated uptake of α-Gal A. Biacore studies revealed that α-Gal A binds to human M6PR with very high affinity, but M6PR also binds to sortilin in a way that prevents α-Gal A binding to sortilin. Taken together, our data provide evidence that sortilin is a new α-Gal A receptor expressed in renal endothelial cells and that this receptor together with the M6PR is able to internalize circulating α-Gal A during enzyme replacement therapy in patients with Fabry disease

Characterisation and treatment of a mouse model of Fabry disease

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Response of Olympia oysters (Ostrea lurida) to changing environmental conditions

Thesis (Master's)--University of Washington, 2014-12The Olympia oyster is an iconic oyster species in the Pacific Northwest with special significance in Puget Sound, WA. Oyster populations in the region were decimated to historic lows during the 20th century due to a number of factors including overharvest, habitat loss, and invasive species. Restoration projects have seen limited success, likely due to the limited information on stock structure within Puget Sound, especially in regards to adaptive abilities and habitat suitability. Chapter one of this study investigates population related fitness measures (ie. mortality, growth, reproduction) within three resident populations from geographically isolated locations in Puget Sound. Using a reciprocal transplant experiment with Ostrea lurida populations from Fidalgo Bay, Dabob Bay, and Oyster Bay, we found that two of the three populations (Dabob Bay and Oyster Bay) express significant phenotypic signatures related to the population. Using this information we offer restoration strategies catered to population phenotypes in an effort to improve restoration projects in the Puget Sound. In Chapter two, we ran a thermal and mechanical stress experiment due to differences in mortality between populations observed in Chapter 1 to investigate expression of genes (via qPCR) related to survival. We found differences in expression related to gene transcription, which indicates possible phenotypic plasticity previously unknown in the study populations though further investigation is required

Professional Learning Communities in elementary schools and how technologies are utilized

Professional Learning Communities (PLC) are being developed by many K–12 public schools, in the year 2010. PLC consist of collaborative teacher teams that focus on evidence of student learning to guide a cycle of instructional improvement. Information and Communications Technologies (ICT) such as those used for assessment, communications and collaboration can facilitate the time consuming work of PLC, and help leaders monitor and support the work of PLC teams. However, educators underutilize ICT. This is the problem examined in the current study. A descriptive and exploratory case study method (Yin, 2009) was used, guided by the conceptual framework of Artifact Analysis (Halverson, 2004). Through interviews, observations and examination of archival data, the research documented how PLC have been enacted in two elementary schools and how technology is used in PLC work. Participants in the study found PLC work to be valuable, stating that it helped them better meet the instructional needs of students and improved their sense of confidence as teachers. Interviews and observations revealed that PLC work is time consuming, adding to an already demanding work schedule. Creative strategies were used to make time for teachers to meet. Participants found assessment data collection and management especially challenging and expressed interest in learning how to make better use of assessment technologies. Participants also expressed an interest in use of ICT to find and share targeted learning activities. The study suggests that communication; assessment and sharing are the areas in which ICT can best facilitate PLC work at this time. Communication technologies can help leaders coordinate the day to day running of the school. Assessment technologies can facilitate the creation, scoring, sharing and analysis of student assessments. Sharing technologies can help teachers find targeted learning activities, and share resources created by team members. The study suggests that ICT tools might be better used if developers seek input from practicing teachers and principals to assess current needs and understand how products will be used in practice. ICT tools should be easy to use in a busy classroom and school environment. Job-embedded professional development is essential when new programs are implemented