Antisense PMO cocktails effectively skip dystrophin exons 45-55 in myotubes transdifferentiated from DMD patient fibroblasts

Antisense-mediated exon skipping has made significant progress as a therapeutic platform in recent years, especially in the case of Duchenne muscular dystrophy (DMD). Despite FDA approval of eteplirsen-the first-ever antisense drug clinically marketed for DMD-exon skipping therapy still faces the significant hurdles of limited applicability and unknown truncated protein function. In-frame exon skipping of dystrophin exons 45-55 represents a significant approach to treating DMD, as a large proportion of patients harbor mutations within this "hotspot" region. Additionally, patients harboring dystrophin exons 45-55 deletion mutations are reported to have exceptionally mild to asymptomatic phenotypes. Here, we demonstrate that a cocktail of phosphorodiamidate morpholino oligomers can effectively skip dystrophin exons 45-55 in vitro in myotubes transdifferentiated from DMD patient fibroblast cells. This is the first report of substantive exons 45-55 skipping in DMD patient cells. These findings help validate the use of transdifferentiated patient fibroblast cells as a suitable cell model for dystrophin exon skipping assays and further emphasize the feasibility of dystrophin exons 45-55 skipping in patients

Studying the variability in patient inflow and staffing trends on Sundays versus other days in the academic emergency department

Background: Resource limitation, staff deficiency, and variability in patient inflow contribute to emergency department (ED) overcrowding, associated with delayed care, poor care, and poor patient outcomes. This study seeks to describe and analyze patient inflow variability and staffing trends on Sundays versus other days in a tertiary academic ED from South India. Methods: Patient inflow and staffing data for 2 years were collected from hospital records, cross-checked, and statistically analyzed using Epi Info 7.0. Results: Significant increase in patient inflow (45.6%) was noted on Sundays compared to other days (155.9 [95% confidence interval (CI): 152.75–159.05] vs. 107.1 [95% CI: 105.98–108.22]; P< 0.001), with higher inflow in the morning shifts (67.4 [95% CI: 65.41–69.45] vs. 32.1 [95% CI: 31.45–32.70]; P< 0.001). All categories of ED staff were deficient across all shifts (2.1 [95% CI: 2.05–2.15] tier-2 physicians, 4.9 [95% CI: 4.86–4.94] nurses, and 1.9 [95% CI: 1.88–1.92] nurse assistants on an average), especially tier-1 physicians (0.3 [95% CI: 0.24–0.36] on Sundays and 0.5 [95% CI: 0.48–0.52] on other days; P< 0.001). Patient-per-hour (PPH)-per-provider based on patient arrival rate was generally high. PPH per tier-1 physician was the highest, being 10.6 (95% CI: 9.95–11.14) versus 5.4 (95% CI: 5.26–5.59; P< 0.001) in the morning and 7.2 (95% CI: 6.95–7.45) versus 6.6 (95% CI: 6.43–6.74; P = 0.08) in the evening shifts on Sundays and other days, respectively. Conclusions: There were deficiencies in all categories of ED staff on all days, and this was pronounced on Sundays due to significantly higher patient inflow. Inadequate ED staffing, especially due to a significant dearth of tier-1 physicians is a pointer toward quality compromise in developing EDs. Authors recommend adequate staff deployment in developing EDs for optimum quality care. This should be implemented such that staffing is based on expected patient inflow so that a PPH-per-provider goal of 2.5 is targeted across all shifts