First, Do No Harm: Understanding Primary Care Providers' Perception of Risks Associated With Discussing Weight With Pediatric Patients.

Many health care providers struggle with if- and how-to discuss weight with their pediatric patients. This study used one-on-one interviews with primary care providers (n = 20) to better understand their: (1) perception of risks associated with talking about weight with pediatric patients, (2) commitment to adhering to best practices of pediatric weight management, and (3) approaches to mitigate perceived risks. Providers felt concerned that discussing weight with children during clinic visits may have unintended negative impacts. Despite perceived risks, providers continued regular BMI screening and weight-focused conversations, but took care with regard to language and approach with the goal of mitigating perceived risks. Findings suggest that pediatric primary care providers perceive that engaging in weight-related discussions with their patients has the potential to lead to negative, unintended consequences. Future research is needed to understand if weight-focused conversations should be avoided altogether or if there are approaches that can effectively mitigate risks.

Primary care providers' perspectives on initiating childhood obesity conversations: a qualitative study.

BACKGROUND: Primary care physicians (PCPs) are in a critical position for identifying, preventing and treating childhood obesity. However, a one-size-fits-all approach does not exist for having conversations about weight with families. A better understanding of how PCPs can address paediatric patients' weight concerns is needed in order to develop effective guidelines and trainings. OBJECTIVE: To describe PCPs preferences and behaviours regarding weight-related conversations with paediatric patients' and their families. METHODS: Twenty PCPs affiliated with the University of Minnesota, USA, were recruited to participate in semi-structured interviews. Transcripts were analysed using inductive thematic analysis. RESULTS: PCP's identified well-child visits as the most appropriate time for weight-related discussions with families. Physicians described what approaches/elements they perceived to work best during conversations: collaboration, empathy, health-focused and objective measures. CONCLUSIONS: Overall, PCPs were more comfortable with weight-related discussions during annual well-child visits and rarely initiated them during an acute visit or the first encounter with a patient. Objective measures, such as growth charts, were often utilized to start discussions. Considering a large proportion of well-child visits are missed, alternative opportunities to have discussions about healthy lifestyle behaviours should be explored. The integral role PCPs play in paediatric obesity warrants further research.

Efficient expression screening of human membrane proteins in transiently transfected Human Embryonic Kidney 293S cells.

It is often an immense challenge to overexpress human membrane proteins at levels sufficient for structural studies. The use of Human Embryonic Kidney 293 (HEK 293) cells to express full-length human membrane proteins is becoming increasingly common, since these cells provide a near-native protein folding and lipid environment. Nevertheless, the labor intensiveness and low yields of HEK 293 cells and other mammalian cell expression systems necessitate the screening for suitable expression as early as possible. Here we present our methodology used to generate constructs of human membrane proteins and to rapidly assess their suitability for overexpression using transiently transfected, glycosylation-deficient GnT I-HEK 293 cells (HEK 293S). Constructs, in the presence or absence of a C-terminal enhanced green fluorescence protein (EGFP) molecule, are made in a modular manner, allowing for the rapid generation of several combinations of fusion tags and gene paralogues/orthologues. Solubilization of HEK 293S cells, using a range of detergents, followed by Western blotting is performed to assess relative expression levels and to detect possible degradation products. Fluorescence-detection size exclusion chromatography (FSEC) is employed to assess expression levels and overall homogeneity of the membrane proteins, to rank different constructs for further downstream expression trials. Constructs identified as having high expression are instantly suitable for further downstream large scale transient expression trials and stable cell line generation. The method described is accessible to all laboratory scales and can be completed in approximately 3 weeks.