An investigation of the early molecular regulation of adipogenesis in human mesenchymal stem cells.

Human bone marrow-derived mesenchymal stem cells (hMSCs) can differentiate in vitro into various lineages including adipocytes, osteoblasts, and chondrocytes. Determining the earliest events of hMSC differentiation would provide a greater understanding of stem cell function, leading to novel therapeutic strategies. The molecular regulation of adipogenesis has been studied extensively using preadipocyte models that are committed to the adipocyte lineage. However, the events that occur in uncommitted stem cells such as hMSCs, and thus control the early stages of differentiation, have not been identified. In this thesis, hMSC differentiation to adipocytes was induced using a classic induction cocktail, and transcriptional profiles were obtained at early timepoints using Affymetrix U133 Plus 2.0 microarrays. Adipogenesis was shown to involve the regulation of over 4200 genes, which were clustered into distinct temporal patterns of gene expression. Recognised adipogenic regulators were expressed at the later stages of adipogenesis alongside genes involved in lipid metabolism, thus demonstrating the validity of the system. Global annotation of genes showing early expression changes revealed temporal regulation of signaling pathways known to control adipogenesis, together with molecular processes and pathway regulators not previously implicated in adipogenesis. A further detailed analysis of gene function identified 83 potential candidates for novel adipogenic regulators. Knock-down of gene expression induced by virally-mediated shRNA is an effective method to assess gene function. To investigate whether the candidate genes identified from the transcriptional profiling played a role in hMSC adipogenesis, retroviral shRNA library screens and an additional lentiviral shRNA strategy were developed and implemented. Finally, the effect on hMSC differentiation of the introduction of oncogenic regulators was investigated. Oncogenic transformation of hMSCs may lead to aberrant differentiation and mesenchymal tumour formation. It was found that the progressive addition of transforming genes did not block adipogenic or osteogenic differentiation, but caused significant impairment to the process

Students’ views on the need for hostile environment awareness training for South African emergency medical care students

Background. South Africans experience high levels of contact crime, including assault, robbery and hijacking. Emergency medical services (EMS) are frequently called to the scene of such incidents. Their presence in these potentially hostile environments increases the risk of South African (SA) paramedics and paramedic students becoming victims of crime and violence. A 2015 study showed that ~66% of SA EMS staff reported being assaulted while on duty. During a 10-month period in 2016, Western Cape Province recorded >40 incidents of physical violence against their EMS personnel. Questions are being asked about how well prepared EMS staff are to operate in potentially hostile environments.Objectives. To explore emergency medical care (EMC) students’ experiences of violence and crime and their views on the need for hostile environment awareness training (HEAT) as part of their undergraduate degree programmes.Methods. A self-designed, non-validated, cross-sectional online survey questionnaire was used to document the experiences, views and opinions of 113 undergraduate EMC students from 4 SA universities. The questionnaire consisted of 24 closed-ended questions with pre-set Likert scale options focusing on documenting participants’ experiences, views and opinions regarding hostile environments and the need for HEAT.Results. A high percentage of participants (92%) indicated feeling unsafe while engaging in clinical learning shifts; 63% specified that they had personally witnessed violence against EMS crews, and 32% indicated that they had been assaulted while on duty. Unsurprisingly, 81% of the respondents felt that there is a need for inclusion of HEAT in the undergraduate curriculum.Conclusions. Participation in clinical learning shifts in the current SA prehospital EMS environment increases the risk of exposure to potentially hostile environments. Consequently, students feel unsafe and support the inclusion of HEAT as part of their undergraduate degree programmes

An adapted triage tool (ETAT) at Red Cross War Memorial Children’s Hospital Medical Emergency Unit, Cape Town: An evaluation

Objective. To evaluate the efficacy of an adapted Emergency Triage Assessment and Treatment (ETAT) tool at a children’s hospital. Design. A two-armed descriptive study. Setting. Red Cross War Memorial Children’s Hospital, Cape Town, South Africa. Methods. Triage data on 1 309 children from October 2007 and July 2009 were analysed. The number of children in each triage category (red (emergency), orange (urgent or priority) and green (non-urgent)) and their disposal were evaluated. Results. 1. The October 2007 series: 902 children aged 5 days - 15 years were evaluated. Their median age was 20 (interquartile range (IQR) 7 - 50) months, and 58.8% (n=530) were triaged green, 37.5% (n=338) orange and 3.8% (n=34) red. Over 90% of children in the green category were discharged (478/530), while 32.5% of children triaged orange (110/338) and 52.9% of children triaged red (18/34) were admitted. There was a significant increase in admission rate for each triage colour change from green through orange to red after adjustment for age category (risk ratio (RR) 2.6; 95% confidence interval (CI) 2.2 - 3.1). 2. The July 2009 cohort: 407 children with a median age of 22 months (IQR 7 - 53 months) were enrolled. Twelve children (2.9%) were triaged red, 187 (45.9%) orange and 208 (51.1%) green. A quarter (101/407) of the children triaged were admitted: 91.7% (11/12) from the red category and 36.9% (69/187) from the orange category were admitted, while 89.9% of children in the green category (187/208) were discharged. After adjusting for age category, admissions increased by more than 300% for every change in triage acuity (RR 3.2; 95% CI 2.5 - 4.1). Conclusions. The adapted ETAT process may serve as a reliable triage tool for busy paediatric medical emergency units in resource-constrained countries and could be evaluated further in community emergency settings

An adapted triage tool (ETAT) at Red Cross War Memorial Children’s Hospital Medical Emergency Unit, Cape Town: An evaluation

Objective. To evaluate the efficacy of an adapted Emergency Triage Assessment and Treatment (ETAT) tool at a children’s hospital. Design. A two-armed descriptive study. Setting. Red Cross War Memorial Children’s Hospital, Cape Town, South Africa. Methods. Triage data on 1 309 children from October 2007 and July 2009 were analysed. The number of children in each triage category (red (emergency), orange (urgent or priority) and green (non-urgent)) and their disposal were evaluated. Results. 1. The October 2007 series: 902 children aged 5 days - 15 years were evaluated. Their median age was 20 (interquartile range (IQR) 7 - 50) months, and 58.8% (n=530) were triaged green, 37.5% (n=338) orange and 3.8% (n=34) red. Over 90% of children in the green category were discharged (478/530), while 32.5% of children triaged orange (110/338) and 52.9% of children triaged red (18/34) were admitted. There was a significant increase in admission rate for each triage colour change from green through orange to red after adjustment for age category (risk ratio (RR) 2.6; 95% confidence interval (CI) 2.2 - 3.1). 2. The July 2009 cohort: 407 children with a median age of 22 months (IQR 7 - 53 months) were enrolled. Twelve children (2.9%) were triaged red, 187 (45.9%) orange and 208 (51.1%) green. A quarter (101/407) of the children triaged were admitted: 91.7% (11/12) from the red category and 36.9% (69/187) from the orange category were admitted, while 89.9% of children in the green category (187/208) were discharged. After adjusting for age category, admissions increased by more than 300% for every change in triage acuity (RR 3.2; 95% CI 2.5 - 4.1). Conclusions. The adapted ETAT process may serve as a reliable triage tool for busy paediatric medical emergency units in resource-constrained countries and could be evaluated further in community emergency settings

Limits to tDCS effects in language:failures to modulate word production in healthy participants with frontal or temporal tDCS

Transcranial direct current stimulation (tDCS) is a method of non-invasive brain stimulation widely used to modulate cognitive functions. Recent studies, however, suggests that effects are unreliable, small and often non-significant at least when stimulation is applied in a single session to healthy individuals. We examined the effects of frontal and temporal lobe anodal tDCS on naming and reading tasks and considered possible interactions with linguistic activation and selection mechanisms as well possible interactions with item difficulty and participant individual variability. Across four separate experiments (N, Exp 1A = 18; 1B = 20; 1C = 18; 2 = 17), we failed to find any difference between real and sham stimulation. Moreover, we found no evidence of significant effects limited to particular conditions (i.e., those requiring suppression of semantic interference), to a subset of participants or to longer RTs. Our findings sound a cautionary note on using tDCS as a means to modulate cognitive performance. Consistent effects of tDCS may be difficult to demonstrate in healthy participants in reading and naming tasks, and be limited to cases of pathological neurophysiology and/or to the use of learning paradigms