Investigating the use of oligonucleotides for the treatment of muscular dystrophy

Antisense oligonucleotide therapy is one of the most promising strategies for treatment of myotonic dystrophy type 1 (DM1), type 2 (DM2) and Duchene muscular dystrophy (DMD). These three diseases have nuclear retained mRNA, suitable for antisense therapy targeting. The delivery of oligonucleotides to their desired target has long been an obstacle in antisense therapy with a large number of delivery reagents or methods having adverse side effects. Promising work published detailing the successful delivery of various chemically modified oligonucleotides (CMOs) naked, via gymnosis, led to us investigating a number of these CMOs: deoxyribonucleic acids (DNA), Peptide nucleic acids (PNAs), 2’OMethyl (2’OMe), and Phosphorodiamidate morpholino (PMO) oligonucleotides. In DM1 expanded CUG repeat (CUGexp) mRNAs aggregate in the nucleus forming “foci”. Testing the CMOs effectiveness at disrupting nuclear foci in a cell based assay, using high content imaging to visualise the number, size and intensity of foci we initially discovered that PNA and 2’OMe, were seemingly taken up via gymnosis by DM1 cells, and removed nuclear foci at nanomolar concentrations. However further experimentation using live cell imaging indicated that although all CMOs could enter the cell, in all disease models tested, via gymnosis, the CMOs could not penetrate the nuclear membrane. In depth analysis led us to identify an artefact of the in-situ process used to identify these foci, explaining earlier positive results. As the target mRNA is trapped within the nuclear compartment we investigated several transfection reagents for their ability to deliver 2’OMe oligonucleotides to the nucleus using live cell fluorescent imaging and a modified northern blot based method. It was established that polyethylenimine could successfully deliver 2’OMe oligonucleotides to the cell, with a high abundance of the oligonucleotide residing within the nuclear compartment. It was observed that PEI degrades the expanded nuclear retained repeat in the DMPK transcript of a DM1 patient cell line alone, without the addition of an antisense agent, in a concentration dependent manner

Prepared to Respond? Investigating Preservice Teachers’ Perceptions of their Readiness for Culturally Responsive Teaching

Increasing cultural diversity in American schools has made preparing graduates to work with diverse learners an essential goal of teacher education programs. The purpose of this study was to investigate the perceptions of elementary (K-6) undergraduate preservice teachers (n=36) in a dual certification program regarding their personal and professional readiness for culturally responsive teaching. Data collected using the Cultural Responsive Teaching Readiness Scale (Karatas & Oral, 2017) revealed relatively high mean overall with little variation (M = 4.25, SD = 0.18) with a statistically significant difference (t (33) = 18.65, p \u3c .001) between Personal Readiness (M = 4.46) and Professional Readiness (M = 3.98). Participants seemed to perceive classroom practice as influential in terms of their cultural awareness, thus, indicating the importance of partnerships with area schools and districts to ensure effective field experiences for preparing teachers for sustained employment in culturally and linguistically diverse settings

3D immersive collection and teaching environments: the Yellow House project at UOW

This paper discusses the Yellow House VR project at the University of Wollongong. Innovative virtual reality technologies such as Oculus Rift are being utilised to recreate the 1970s Sydney artist community space known as the Yellow House, as both an historic replication and openly accessible, immersive teaching and learning environment for use and adaptation by teachers, students, researchers and the general community. The paper considers the role of the library in the enhanced presentation of digitised collections through new and evolving technologies that provide opportunities for knowledge enhancement and support the development of student e-portfolios

Making A Difference: Year Two Report of the Pennsylvania High School Coaching Initiative

This report examines the implementation of the second year of three for the Pennsylvania High School Coaching Initiative (PAHSCI). Funded by the Annenberg Foundation, this initiative focuses on literacy and math coaches providing support to teachers from across the major subject areas to create literacy-rich classrooms in which students actively engage in learning tasks that deepen their content knowledge and strengthen their abilities to think critically and communicate well. This report presents findings from the first two years of research. It includes survey research as well as in-depth qualitative research in participating schools and districts and provides recommendations for PAHSCI stakeholders as they refine the program and for other education reformers as they consider the benefits of instructional coaching as a strategy for improving high schools and student achievement

Impacts of added roughage on growth performance, digestibility, ruminal fermentation, and ruminal pH of feedlot steers fed wheat-based feedlot diets containing 30% modified distillers grains with solubles

Two experiments were conducted to evaluate the inclusion rate roughage in wheat-based diets containing modified distillers grains with solubles (MDGS) on feedlot performance (Feedlot Experiment), as well as digestibility, ruminal pH, and ruminal fermentation characteristics (Digestibility Experiment). The feedlot experiment utilized 72 Angus steers (392 ± 46.3 kg initial body weight) which were randomly assigned to 1 of 12 pens, 3 pens per treatment, to evaluate feedlot performance and carcass characteristics. Dietary treatments were 1) control; 10% roughage, 2) 12% roughage, 3) 14% roughage, and 4) 16% roughage. The digestibility experiment used four ruminally and duodenally cannulated steers (393 ± 33.0 kg) in a 4 × 4 Latin Square with either 10%, 12%, 14%, or 16% roughage as in the feedlot experiment. However, dietary roughage source was different between these two experiments and included a combination of grass hay and wheat straw (Feedlot Experiment), and corn silage (Digestibility Experiment). All data were analyzed with the Mixed Procedures of SAS. Feed intake was recorded, with duodenal and fecal output calculated using chromic oxide. Ruminal pH and fermentation were assessed. Growth performance and most carcass characteristics were not affected by increasing roughage (P ≥ 0.11). Marbling tended to decrease linearly (P = 0.10) with increasing roughage inclusion. Increasing dietary roughage content had no effect on organic matter intake (P = 0.60) in the digestibility experiment. Intake, duodenal flow, and digestibility of neutral detergent fiber and acid detergent fiber were not affected by treatment (P ≥ 0.16). Ruminal pH increased linearly (P \u3c 0.01) as rate of roughage inclusion increased. Ruminal concentrations of acetate and butyrate increased, and propionate decreased in a linear fashion (P \u3c 0.01) thereby increasing (P \u3c 0.01) acetate and butyrate to propionate ratio with increasing dietary roughage. Our data indicate that increasing roughage inclusion in wheat-based diets including 30% MDGS increased ruminal pH and shifted ruminal fermentation patterns. Additionally, increasing roughage inclusion did not affect feedlot performance in steers fed wheat at 36% to 42% of dietary dry matter in combination with 30% MDGS

Analyzing Commercial Video Game Instruction through the Lens of Instructional Design

This paper will examine how Gagne’s Nine Events of Instruction (1992) may appear, perhaps inadvertently, within commercial games that guide the user from novice to expert player. By employing a qualitative artifact analysis methodology, we examine a popular action adventure video game to determine if game designers encourage players to build game expertise by employing similar events to Gagne’s instructional design model. We demonstrate that our artifact of analysis does consistently employ Gagne’s events, though often in a manner unique to a digitally mediated space. We conclude that an experiential game setting has the potential to be a platform for instructional delivery

Practical and emotional preparation for death: A mixed methods study investigating experiences of family carers of people with dementia

BACKGROUND: When family carers are more prepared for the end of the life of a person they care for, they report improved bereavement outcomes. Few studies have explored how carers prepare for the death of a person with dementia. We aimed to explore how carers for people with all stages of dementia experience preparing for end of life care and death. METHODS: This was a mixed methods cross-sectional study. Family carers of people with dementia (n = 150) completed a structured interview with validated scales, alongside questions about death preparedness and advance decisions. A sub-sample (n = 16) completed qualitative interviews exploring their experiences of planning for end of life. We fitted logistic regression models to explore associations with preparedness, and thematically analysed qualitative data. RESULTS: We addressed practical and emotional preparation separately for 143 participants. Fifty seven percent of participants were very practically prepared for death, while only 29% were very emotionally prepared. Male carers were more likely than female carers to report being very emotionally and practically prepared. Higher engagement with healthcare professionals was associated with feeling very practically prepared; although we found that formal discussions of end of life care issues with healthcare professionals did not impact carers’ feelings of preparation. Higher levels of dementia severity and carer depression were associated with feeling very emotionally prepared. Three qualitative themes related to practical and emotional preparation were identified: (1) ambiguity and uncertainty; (2) support from the system; and (3) how death is perceived by the carer. CONCLUSIONS: While most carers felt practically prepared for death, emotional preparation was much lower. Further research is needed to understand how engagement with healthcare professionals or other forms of social or emotional support could help carers, particularly female carers, to emotionally prepare for their relative’s death

Practical and emotional preparation for death: A mixed methods study investigating experiences of family carers of people with dementia

BACKGROUND: When family carers are more prepared for the end of the life of a person they care for, they report improved bereavement outcomes. Few studies have explored how carers prepare for the death of a person with dementia. We aimed to explore how carers for people with all stages of dementia experience preparing for end of life care and death. METHODS: This was a mixed methods cross-sectional study. Family carers of people with dementia (n = 150) completed a structured interview with validated scales, alongside questions about death preparedness and advance decisions. A sub-sample (n = 16) completed qualitative interviews exploring their experiences of planning for end of life. We fitted logistic regression models to explore associations with preparedness, and thematically analysed qualitative data. RESULTS: We addressed practical and emotional preparation separately for 143 participants. Fifty seven percent of participants were very practically prepared for death, while only 29% were very emotionally prepared. Male carers were more likely than female carers to report being very emotionally and practically prepared. Higher engagement with healthcare professionals was associated with feeling very practically prepared; although we found that formal discussions of end of life care issues with healthcare professionals did not impact carers' feelings of preparation. Higher levels of dementia severity and carer depression were associated with feeling very emotionally prepared. Three qualitative themes related to practical and emotional preparation were identified: (1) ambiguity and uncertainty; (2) support from the system; and (3) how death is perceived by the carer. CONCLUSIONS: While most carers felt practically prepared for death, emotional preparation was much lower. Further research is needed to understand how engagement with healthcare professionals or other forms of social or emotional support could help carers, particularly female carers, to emotionally prepare for their relative's death

Effects of Marsh Edge Erosion in Coupled Barrier Island-Marsh Systems and Geometric Constraints on Marsh Evolution

Sand washed across barrier islands during storms (called overwash) provides sediment for salt marshes behind those islands, and can allow a marsh which otherwise would drown to grow vertically fast enough to keep up with sea level. We use a barrier island-marsh evolution model (GEOMBEST+) to see what effect marsh edge erosion by waves has on overwash-supported marshes. Consistent with previous research, we find that wave erosion can make marshes more resilient by freeing sediment that can be used elsewhere on the marsh surface. We add that horizontal erosion of the marsh edge provides more sediment per volume eroded than vertical erosion of the marsh surface. This is because the bottom layers of the marsh contain more sediment (that can stay on marsh surfaces), while the surface layers include plant material (that drifts away or decomposes). We also find that when the marsh and bay are keeping up with sea level, expanding or eroding the marsh is the only way to change the volume of the bay, so how fast the marsh is expanding or eroding can be predicted using geometry, knowing only the size of the basin, sea-level-rise rate, and the net rate of sediment import or export

Drinking-Water Arsenic Exposure Modulates Gene Expression in Human Lymphocytes from a U.S. Population

Background: Arsenic exposure impairs development and can lead to cancer, cardiovascular disease, and diabetes. The mechanism underlying these effects remains unknown. Primarily because of geologic sources of contamination, drinking-water arsenic levels are above the current recommended maximum contaminant level of 10 μg/L in the northeastern, western, and north central regions of the United States. Objectives: We investigated the effects of arsenic exposure, defined by internal biomarkers at levels relevant to the United States and similarly exposed populations, on gene expression. Methods: We conducted separate Affymetrix microarray-based genomewide analyses of expression patterns. Peripheral blood lymphocyte samples from 21 controls interviewed (1999–2002) as part of a case–control study in New Hampshire were selected based on high- versus low-level arsenic exposure levels. Results: The biologic functions of the transcripts that showed statistically significant abundance differences between high- and low-arsenic exposure groups included an overrepresentation of genes involved in defense response, immune function, cell growth, apoptosis, regulation of cell cycle, T-cell receptor signaling pathway, and diabetes. Notably, the high-arsenic exposure group exhibited higher levels of several killer cell immunoglobulin-like receptors that inhibit natural killer cell activity. Conclusions: These findings define biologic changes that occur with chronic arsenic exposure in humans and provide leads and potential targets for understanding and monitoring the pathogenesis of arsenic-induced diseases