Mutagenesis of Human Alpha-Galactosidase A for the Treatment of Fabry Disease

Fabry disease is an X-linked lysosomal storage disorder caused by the deficiency of the enzyme, α-galactosidase A, which results in the accumulation of the lipid substrate. This accumulation results in obstruction of blood flow in patients and early demise at approximately 40-60 years of age. There is currently only one FDA approved treatment (Fabrazyme) classified as an enzyme replacement therapy. However, approximately 88% of patients experience a severe immune response that, rarely, can be fatal and is a huge cost burden at average $250,000 a year per patient. The structure of α-galactosidase A has been previously determined to be a homodimer with six N-linked glycosylation sites, and the catalytic mechanism determined to be a ping-pong bi-bi with the second substrate being water. The purpose of this research is twofold: first, to generate a more efficient enzyme replacement therapy alternative; and second, to develop an alternative assay to detect the activity of the enzyme that can lead to a better screen for the presence of lysosomal storage disorders in patients. A more efficient therapy was investigated utilizing two different host expression systems, Escherichia coli and Pichia pastoris, as well as site directed mutagenesis of the human enzyme. Detectable expression was not observed in E. coli, so mutagenesis was carried out using P. pastoris. Three locations on the structure of α-galactosidase A protein were targeted for mutation the active site, the dimer interface, and the hydrophobic loop between the active site and the third glycosylation site. v Viable candidates for further study into a therapeutic α-galactosidase A were determined based on catalytic efficiency (kcat/Km), thermostability, and possible glycosylation independence. Three mutations were identified to be of potential therapeutic significance in each one of the targeted areas. A continuous enzyme assay was developed with an artificial fluorescent substrate, 4-methylumbelliferyl-α-D-galactopyranoside, that at acidic pH generates the same kinetic values as the preexisting discontinuous time point assay at basic pH. This development has not only potential laboratory benefits if implemented; it also has a clinical significance in screening for Fabry disease and can potentially be extended to other lysosomal storage disorders

Effects of Original XPC on Newly Weaned Beef Steer Nutrient Digestibility and Response to a Vaccination Challenge

The study was designed to determine the effects of Diamond V Original XPC, a yeast fermentation product, in the diets of newly weaned beef steers on nutrient digestibility and response to a vaccination challenge. Although no overall performance benefit was noted, XPC improved total tract CP digestibility. Steers fed XPC at 14 g/d exhibited lesser concentrations of APP, greater DMI, and more efficient rumination post-vaccination. Further research is needed to determine the optimal supplementation rate of XPC to newly received beef cattle

Effects of Original XPC on Newly Weaned Beef Steer Growth Performance and Antioxidant Defense

The objective of this study was to determine the effects of increasing inclusions of Diamond V Original XPC, a yeast fermentation product, on newly weaned beef steer performance and antioxidant defense. There was no effect of Original XPC on growth performance in the current study; however, blood measures indicated greater antioxidant capacity and lesser oxidative stress for steers fed XPC at 14 g/d. Further research is needed to better understand how oxidative stress impacts animal performance and health as well as the optimum supplementation dose of XPC for newly received beef cattle

Living with Traumatic Brain Injury in a Rural Setting: Supports and Barriers Across the Continuum of Care

Purpose: Traumatic brain injury (TBI) is prevalent in Kentucky and comes with a high cost in care and quality of life for individuals and caregivers affected. Many people living with the condition of TBI have unmet needs. Research among people living with TBI in rural areas is limited. The purposes of this study were to (1) increase understanding of the lived experience of people with TBI and caregivers in rural regions of Kentucky across the continuum of their care and (2) provide their perspectives on barriers and facilitators of optimal function and well-being. Methods: A qualitative descriptive interview study was conducted by a multidisciplinary team. Content analysis was completed with data-derived coding and iterative modifications to analysis, coalescing codes into categories and themes. Results: Thirteen people with TBI and six caregivers participated in the interview. Categories that emerged in analysis included the experiences under each locus of care; themes included relationships, functional competence, and participation in meaningful activity. Conclusion: Relationships represented both barriers and facilitators of well-being. Major unmet needs persisted in terms of medical problems, support for caregivers, community linkages, and participation in meaningful activities. Recommendations are made regarding avenues for addressing unmet needs

Teacher Perceptions of Skills, Knowledge, and Resources Needed to Promote Social and Emotional Learning in Rural Classrooms

The incorporation of social and emotional learning (SEL) in schools has been shown to improve academic and psychological health of students. Research has been limited regarding implementation of SEL programs in rural communities, where student needs are heightened. The current study examined factors that could impact teachers’ intentions to be early adopters of a SEL curriculum in a rural community. Seventy-six teachers provided self-report data regarding perceptions of professional strengths, school climate, school resources for student support, ability to educate diverse students, ability to teach specific SEL domains, and intentions to be an early adopter of a SEL program. Present results indicated positive perceptions of school climate, one’s ability to teach diverse students, and one’s ability to teach self-management skills positively predicted intentions to be an early adopter of a SEL curriculum. Implications for rural schools are explored and recommendations for adoption of SEL curricula in rural schools are provided

Discovery and Annotation of Two Phages that Infect Microbacterium foliorum: Tedro and Bajuniper

We isolated and purified Tedro and BAjuniper which infect the host Microbacterium foliorium. Tedro is a lytic, cluster EF phage isolated from soil collected in Hawarden, Iowa. Its genome is 56,197 bp long, circularly permuted, and includes 83 protein-coding genes and no tRNA genes. We are examining two of Tedro’s genes, genes 56 and 57, both of which are predicted to encode a DnaE-like DNA polymerase III (alpha) in more detail. Tedro_57 is twice as large as Tedro_56 so we are using additional bioinformatic tools to understand these genes. BAjuniper was isolated from soil collected in a garden in Orange City, Iowa. Its genome is 41,985 bp long. It was assigned to cluster EB. BAjuniper’s genome includes one tRNA gene and we will finalize BAjuniper’s annotation shortly

Social and Cultural Factors Affecting Uptake of Interventions for Malaria in Pregnancy in Africa: A Systematic Review of the Qualitative Research

Malaria during pregnancy (MiP) results in adverse birth outcomes and poor maternal health. MiP-related morbidity and mortality is most pronounced in sub-Saharan Africa, where recommended MiP interventions include intermittent preventive treatment, insecticide-treated bednets and appropriate case management. Besides their clinical efficacy, the effectiveness of these interventions depends on the attitudes and behaviours of pregnant women and the wider community, which are shaped by social and cultural factors. Although these factors have been studied largely using quantitative methods, qualitative research also offers important insights. This article provides a comprehensive overview of qualitative research on social and cultural factors relevant to uptake of MiP interventions in sub-Saharan Africa.A systematic search strategy was employed: literature searches were undertaken in several databases (OVID SP, IS Web of Knowledge, MiP Consortium library). MiP-related original research, on social/cultural factors relevant to MiP interventions, in Africa, with findings derived from qualitative methods was included. Non-English language articles were excluded. A meta-ethnographic approach was taken to analysing and synthesizing findings. Thirty-seven studies were identified. Fourteen concentrated on MiP. Others focused on malaria treatment and prevention, antenatal care (ANC), anaemia during pregnancy or reproductive loss. Themes identified included concepts of malaria and risk in pregnancy, attitudes towards interventions, structural factors affecting delivery and uptake, and perceptions of ANC.Although malaria risk is associated with pregnancy, women's vulnerability is often considered less disease-specific and MiP interpreted in locally defined categories. Furthermore, local discourses and health workers' ideas and comments influence concerns about MiP interventions. Understandings of ANC, health worker-client interactions, household decision-making, gender relations, cost and distance to health facilities affect pregnant women's access to MiP interventions and lack of healthcare infrastructure limits provision of interventions. Further qualitative research is however required: many studies were principally descriptive and an in-depth comparative approach is recommended

Epidemiology of injuries from fire, heat and hot substances : global, regional and national morbidity and mortality estimates from the Global Burden of Disease 2017 study

Background Past research has shown how fires, heat and hot substances are important causes of health loss globally. Detailed estimates of the morbidity and mortality from these injuries could help drive preventative measures and improved access to care. Methods We used the Global Burden of Disease 2017 framework to produce three main results. First, we produced results on incidence, prevalence, years lived with disability, deaths, years of life lost and disability-adjusted life years from 1990 to 2017 for 195 countries and territories. Second, we analysed these results to measure mortality-to-incidence ratios by location. Third, we reported the measures above in terms of the cause of fire, heat and hot substances and the types of bodily injuries that result. Results Globally, there were 8 991 468 (7 481 218 to 10 740 897) new fire, heat and hot substance injuries in 2017 with 120 632 (101 630 to 129 383) deaths. At the global level, the age-standardised mortality caused by fire, heat and hot substances significantly declined from 1990 to 2017, but regionally there was variability in age-standardised incidence with some regions experiencing an increase (eg, Southern Latin America) and others experiencing a significant decrease (eg, High-income North America). Conclusions The incidence and mortality of injuries that result from fire, heat and hot substances affect every region of the world but are most concentrated in middle and lower income areas. More resources should be invested in measuring these injuries as well as in improving infrastructure, advancing safety measures and ensuring access to care.Peer reviewe