Five head and neck cancer quality metrics were identified that have substantial variability in adherence and meaningfully impact overall survival. These metrics are appropriate candidates for national adoption. Cancer 2017;123:4372-81. © 2017 American Cancer Society.
Muscular dystrophies are a class of disorders that cause progressive muscle wasting. A major hurdle for discovering treatments for the muscular dystrophies is a lack of reliable assays to monitor disease progression in animal models. We have developed a novel mouse model to assess disease activity noninvasively in mice with muscular dystrophies. These mice express an inducible luciferase reporter gene in muscle stem cells. In dystrophic mice, muscle stem cells activate and proliferate in response to muscle degeneration, resulting in an increase in the level of luciferase expression, which can be monitored by noninvasive, bioluminescence imaging. We applied this noninvasive imaging to assess disease activity in a mouse model of the human disease limb girdle muscular dystrophy 2B (LGMD2B), caused by a mutation in the dysferlin gene. We monitored the natural history and disease progression in these dysferlin-deficient mice up to 18 months of age and were able to detect disease activity prior to the appearance of any overt disease manifestation by histopathological analyses. Disease activity was reflected by changes in luciferase activity over time, and disease burden was reflected by cumulative luciferase activity, which paralleled disease progression as determined by histopathological analysis. The ability to monitor disease activity noninvasively in mouse models of muscular dystrophy will be invaluable for the assessment of disease progression and the effectiveness of therapeutic interventions.
Background: Timely and effective bystander first aid can improve outcomes for trauma victims. Bystanders are present at most traumas and are more likely to assist with prior training. Materials and methods: An evidence-based course was created for the general public in highrisk Chicago neighborhoods focused on basic traumatic first aid, including scene management, hemorrhage control, and mitigating the psychological impact of trauma to overcome the bystander effect. Prospectively, participants completed knowledge-based and self-efficacy assessments precourse, postcourse, and 6 mo follow-up. The change in self-efficacy and knowledge scores was analyzed. Results: Over 32 courses, 503 participants were taught; 474 and 460 participants completed precourse and postcourse surveys, respectively, whereas 60 of 327 who consented for follow-up completed the 6-mo survey. Postcourse, participants were more likely to assist trauma victims and felt more confident in the quality of care they could provide; the effect remained significant at 6 mo (all P < 0.001). All seven self-efficacy empowerment-based questions individually demonstrated improvement from precourse to postcourse (P < 0.001), with an overall mean (SD) increase of 2.8 (2.1, P < 0.001); six maintained significance at follow-up with an overall mean increase of 2.8 (1.9, P < 0.001). Knowledge scores improved from 6.2 of 10 to 7.2 postcourse and 7.7 at follow-up (P < 0.001). Most improved were the ability to render first aid and apply tourniquets. Conclusions: The TFRC increased self-efficacy, successfully teaching initial trauma care, particularly hemorrhage control and scene safety, suggesting that a grassroots approach to trauma care may improve outcomes in communities that experience high violence rates.
OBJECTIVE: To test the efficiency of direct spherification in supporting differentiation of mouse embryonic stem cells (mESCs) into meiotic male germ cells.DESIGN: Biological 3D scaffolding was utilized to culture and differentiate mESCs. Two main methods were investigated: 1) mESCs were injected into pre-formed spheres (IN), and 2) a suspension of mESCs was directly encapsulated (DE) into spheres. To coax differentiation, spheres were bathed in a growth factor cocktail. After mechanical breaching of the spheres, serial isolation at different time intervals was carried out. These isolated cells were analyzed with germ cell stage-specific markers.MATERIALS AND METHODS: MESCs were initially cultured on a 6well dish, trypsinized, and resuspended in stem cell medium. For the IN method, the suspension was injected by using a 1 mL syringe into pre-formed spheres. For the DE method, spheres were formed by suspending mESCs in a base spherification solution and encapsulated by exposure to sodium alginate. Four spheres were bathed in EpiLC differentiating medium containing Activin A, bFGF, and KSR for 3 days, and then submerged in spermatogonial stem cell (SSC) medium composed of DMEM, GDNF, FGF2, 2-mercaptoethanol, L-glutamine, B27 supplement, and 1 mM retinoic acid (RA) for 7 days. Each sphere measured at a diameter of 8 to 10 mm, containing approximately 6.3x10⁵ cells per sphere. Cells were isolated and analyzed after 3 days for OCT4 and Nanog, and DAZL and VASA at day 10.RESULTS: Both IN and DE proved successful in supporting the growth of mESCs. However, optic visualization through the membrane of the IN sphere showed a lower concentration compared to the DE approach. Embryoid body formation was first observed in the DE group at day 4, indicating greater efficiency with this approach. Confident with the sustainability and efficiency of DE, we attempted differentiation solely by this method, by bathing the spheres in EpiLC medium. After 3 days of culture, two spheres were breached and their cellular content was analyzed, showing OCT4 expression in about 85% of cells and a more conspicuous decrease in Nanog expression (<40%), indicating successful progression to EpiLCs. The remaining two spheres cultured in SSC medium with RA from day 4 to 10 showed an increase in embryoid body size that even extruded through the sphere wall. At day 10, the spheres were mechanically breached, and isolated cells showed positivity for VASA ($5%) and DAZL ($5%), confirming meiotic differentiation into germ cells.CONCLUSIONS: Successful sustenance and propagation of mESCs using direct spherification indicates the feasibility of this 3D culture system to promote differentiation towards the male germ line. The ability to support differentiation to post-meiotic stages confirmed by biological proof would render this culture method sustainable for in vitro neogametogenesis.
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