Bioengineered niches that recreate physiological extracellular matrix organisation to support long-term haematopoietic stem cells.

Long-term reconstituting haematopoietic stem cells (LT-HSCs) are used to treat blood disorders via stem cell transplantation. The very low abundance of LT-HSCs and their rapid differentiation during in vitro culture hinders their clinical utility. Previous developments using stromal feeder layers, defined media cocktails, and bioengineering have enabled HSC expansion in culture, but of mostly short-term HSCs and progenitor populations at the expense of naive LT-HSCs. Here, we report the creation of a bioengineered LT-HSC maintenance niche that recreates physiological extracellular matrix organisation, using soft collagen type-I hydrogels to drive nestin expression in perivascular stromal cells (PerSCs). We demonstrate that nestin, which is expressed by HSC-supportive bone marrow stromal cells, is cytoprotective and, via regulation of metabolism, is important for HIF-1α expression in PerSCs. When CD34+ve HSCs were added to the bioengineered niches comprising nestin/HIF-1α expressing PerSCs, LT-HSC numbers were maintained with normal clonal and in vivo reconstitution potential, without media supplementation. We provide proof-of-concept that our bioengineered niches can support the survival of CRISPR edited HSCs. Successful editing of LT-HSCs ex vivo can have potential impact on the treatment of blood disorders.

Knowledge and Preparedness of Physicians in Relation to Anaphylaxis at Primary Healthcare Centers in Al-Qassim, Saudi Arabia.

Background Anaphylaxis is a significant, often fatal, systemic allergic reaction with a rapid start that may affect the respiratory and/or circulatory systems; for patients to survive, emergency management must be done properly. When anaphylaxis is confirmed or highly suspected, epinephrine should be injected intramuscularly. This study aimed to assess the preparedness of primary healthcare physicians for anaphylaxis in terms of recognition and management of this condition in Al-Qassim, Saudi Arabia. Methods This cross-sectional study was carried out among primary healthcare physicians in the Qassim region of Saudi Arabia. A self-administered questionnaire was distributed among the targeted physicians. The questionnaire comprised sociodemographic characteristics, general awareness and management of anaphylaxis, and a 10-item questionnaire to assess physicians' knowledge of anaphylaxis. Results Out of 121 primary healthcare physicians, 61.2% were male, and 47.9% were aged between 25 and 35 years. Nearly all physicians (97.5%) believed that anaphylaxis is a life-threatening situation. The overall mean knowledge score was 4.74 out of 10 points. Nearly half (48.8%) were categorized as having poor knowledge levels, 43% as moderate, and only 8.3% as having good knowledge. Being specialists/consultants and being trained in managing anaphylaxis were the factors associated with increased knowledge. Conclusion The knowledge of primary care physicians regarding anaphylaxis was deficient. However, increased knowledge of anaphylaxis was seen more frequently among consultants or specialists who attended training for managing anaphylaxis cases. A multi-center study involving a bigger sample size is needed to establish physicians' knowledge of anaphylaxis.

The Heartfile Lodhran CVD prevention project--end of project evaluation

Mainstream preventive interventions often fail to reach poor populations with a high risk of cardiovascular diseases (CVDs) in Pakistan. A community-based CVD primary prevention project aimed at developing approaches to reduce risk factors in such populations was established by Heartfile in collaboration with the National Rural Support Program in the district of Lodhran. The project implemented a range of activities integrated with existing social and health service mechanisms during a three year intervention period 2000/01-03/04. These were targeted in 4 key settings: community health education, mass media interventions, training of health professionals and health education through Lady Health Workers. The project received support from the Department for International Development, U.K. At the community level, a pre-test-post-test quasi-experimental design was used for examining project outcomes related to the community component of the intervention. Pre and post-intervention (training) evaluations were conducted involving all health care providers in randomly selected workshops in order to determine baseline levels of knowledge and the impact of training on knowledge level. In order to assess practices of physician and non-physician health care providers patient interviews, with control comparisons were conducted at each health care facility. Significant positive changes were observed in knowledge levels at a community level in the district of intervention compared with baseline knowledge levels particularly in relation to a heart healthy diet, beneficial level of physical activity, the causes of high blood pressure and heart attack and the effects of high blood pressure and active and passive smoking on health. Significant changes in behaviors at a practice level were not shown in the district of intervention. However the project played a critical role in spurring national action for the prevention and control of non-communicable diseases and introducing sustainable public health interventions for poor communities in Pakistan. (AU)