Brown Fat Development in Wild-type and NG2 Null Mice.

<p>(A–C) Oil Red O staining of IBAT from P5 wild-type (A) and NG2 null (B) mice and quantification of Oil Red O staining intensity (C, n = 6). Bar = 50 µm. D. mRNA levels for brown fat-related genes in IBAT from P5 wild-type and NG2 null mice (n = 3). (E–H) Oil Red O staining of primary wild-type (E) and NG2 null (F) as well as immortalized wild-type (G) and NG2 null (H) pre-brown fat cells after 5 days of differentiation. Bar = 50 µm. (I–L) Immuno-fluorescence to detect NG2 and PDGFRβ in immortalized wild-type brown pre-adipocytes (I and K) and NG2 null brown pre-adipocytes (J and L). Bar = 100 µm. (M) Messenger RNA levels of PRDM16 and PGC1-α in immortalized pre-brown fat cells from wild-type and NG2 null mice (n = 3). *, P<0.05; **, P<0.01.</p

MEF Adipogenesis and Expression of NG2 in Adipocytes.

<p>(A and B) Oil red O staining to visualize differentiated adipocytes from wild-type (A) and NG2 null (B) MEFs. (C) Percentages of Oil red O stained cells <i>vs</i> total cells were determined in 10 randomly-selected high power fields. Results are quantified as Mean ± SEM. **, P<0.01. (D) Western blot analysis of NG2 protein in extracts of epididymal fat tissue from 3 month old wild-type and NG2 null mice. (E and F) Immunofluorescence labeling of NG2 in 20 µm-thick cryosections of epididymal fat tissue from 3-month old wild-type and NG2 null mice. (G and H) Immunofluorescence labeling of NG2 in 10 µm-thick cryosections of interscapular brown fat tissue from postnatal day 5 wild-type and NG2 null mice. Bar = 50 µm.</p

NG2 Null Mice are obese.

<p>(A and B) Body weight gain in wild-type and NG2 null mice. (C) Body weight and epididymal fat weight (grams) of 16-week old male wild-type and NG2 null mice (n = 6–10). (D) Body weight and weight of gonadal fat deposits in 16 week female mice (n = 7–9). (E) Dissected epididymal fat pads from 16-week old male wild-type and NG2 null mice. Fat pads are shown in 35-mm dishes. (F) Sizes of white fat cells in sections of epididymal fat pads from 16-week old male wild-type and NG2 null mice. Bar = 50 µm. (G) Numbers of fat cells per mm² (n = 5). (H) Expression of NG2 (Red), PDGFRα(Green) and CD31 (Blue) in sections through the ventromedial hypothalamic nucleus in both wild-type and OPC-NG2 Knockout mice. Note that yellow color in WT is due to red/green overlap. NG2 is not expressed by OPCs in the OPC-NG2 null mouse. Bar = 50 µm. (I) Body weight and epididymal fat weight in 16-week old wild-type and OPC-NG2 null mice (n = 8–9). (J and K) GTT (at 16 weeks) in male and Female wild-type and NG2 null mice (n = 5). (L) ITT (at 25 weeks) in male wild-type and NG2 null mice (n = 5). *, p<0.05; **, p<0.01.</p

Metabolic and Cold Challenge Testing in Wild-type and NG2 Null Mice.

<p>(A and B) Volume of O₂ consumption (A) and CO₂ production (B) by 3-month old wild-type and NG2 null mice over 48 hours (Two light/dark cycles, n = 15). Data were normalized to body weights of individual mice. (C) Cold challenge testing of 3-month old male wild-type and NG2 null mice (n = 5). (D) Three month old male wild-type and NG2 null mice were put on HFD for 10 weeks and body temperatures were measured. As a control, body temperatures of five month old male wild-type and NG2 null mice on regular chow were measured (n = 6–7). (E) Levels of mRNA for brown fat-related genes in IBAT of wild-type and NG2 null mice maintained at room temperature or at 4°C for 6 hours (n = 3). *, P<0.05; **, P<0.01.</p

Study booklet for control group.

BackgroundBathing babies less frequently and intensively in the first six months of life may prevent eczema, but this has not yet been definitively tested in a randomised controlled trial. Such a trial would require evidence-based support to help parents engage with a minimal bathing routine. The present study reports the development of this support.MethodsWe adopted a four-stage design process: (i) Pregnant women and their families (n = 31) were interviewed to ascertain key barriers and facilitators towards following the minimal bathing intervention. (ii) These barriers and facilitators were mapped to behaviour change techniques, focussing on the intervention types of education, persuasion and environmental restructuring, alongside appropriate modes of delivery, and prototype intervention materials were developed. (iii) We iteratively refined these materials in a workshop with multidisciplinary experts and Patient and Public Involvement and Engagement (PPIE) representatives (n = 13) and an (iv) intervention walkthrough with families (n = 5). The design process was informed by the Behaviour Change Wheel, Theoretical framework of acceptability and the Template for intervention description and replication.ResultsSocial influences and motivational factors are likely to influence both uptake and adherence to the intervention. Anticipated emotional reward from participating in research for the benefit of others was indicated to be a strong facilitator for intervention uptake. Alternatives to bathing, having fun with the baby and the night-time routine, alongside family support, were notable facilitators suggested to aid adherence to the intervention. Barriers included hygiene concerns and anticipated negative social appraisal. Barriers and facilitators were mapped to thirty-six behaviour change techniques, focussing on the intervention types of education, persuasion and environmental restructuring, all of which were embedded into the package of support. The prototype intervention materials received positive feedback from the expert workshop and study walkthrough with families. The final package of support comprises printed and digital prompts and cues, a study booklet, video, and digital tool for self-monitoring.ConclusionsThe intervention design process incorporated the ‘real world’ views and experiences of families, experts and PPIE representatives, alongside criteria for designing behavioural interventions. The effectiveness of the package of support will be tested in a feasibility trial and embedded process evaluation.</div

Image of control group pack.

BackgroundBathing babies less frequently and intensively in the first six months of life may prevent eczema, but this has not yet been definitively tested in a randomised controlled trial. Such a trial would require evidence-based support to help parents engage with a minimal bathing routine. The present study reports the development of this support.MethodsWe adopted a four-stage design process: (i) Pregnant women and their families (n = 31) were interviewed to ascertain key barriers and facilitators towards following the minimal bathing intervention. (ii) These barriers and facilitators were mapped to behaviour change techniques, focussing on the intervention types of education, persuasion and environmental restructuring, alongside appropriate modes of delivery, and prototype intervention materials were developed. (iii) We iteratively refined these materials in a workshop with multidisciplinary experts and Patient and Public Involvement and Engagement (PPIE) representatives (n = 13) and an (iv) intervention walkthrough with families (n = 5). The design process was informed by the Behaviour Change Wheel, Theoretical framework of acceptability and the Template for intervention description and replication.ResultsSocial influences and motivational factors are likely to influence both uptake and adherence to the intervention. Anticipated emotional reward from participating in research for the benefit of others was indicated to be a strong facilitator for intervention uptake. Alternatives to bathing, having fun with the baby and the night-time routine, alongside family support, were notable facilitators suggested to aid adherence to the intervention. Barriers included hygiene concerns and anticipated negative social appraisal. Barriers and facilitators were mapped to thirty-six behaviour change techniques, focussing on the intervention types of education, persuasion and environmental restructuring, all of which were embedded into the package of support. The prototype intervention materials received positive feedback from the expert workshop and study walkthrough with families. The final package of support comprises printed and digital prompts and cues, a study booklet, video, and digital tool for self-monitoring.ConclusionsThe intervention design process incorporated the ‘real world’ views and experiences of families, experts and PPIE representatives, alongside criteria for designing behavioural interventions. The effectiveness of the package of support will be tested in a feasibility trial and embedded process evaluation.</div

Consolidated criteria for reporting qualitative research (COREQ) items.

Consolidated criteria for reporting qualitative research (COREQ) items.</p

Study booklet for intervention group.

BackgroundBathing babies less frequently and intensively in the first six months of life may prevent eczema, but this has not yet been definitively tested in a randomised controlled trial. Such a trial would require evidence-based support to help parents engage with a minimal bathing routine. The present study reports the development of this support.MethodsWe adopted a four-stage design process: (i) Pregnant women and their families (n = 31) were interviewed to ascertain key barriers and facilitators towards following the minimal bathing intervention. (ii) These barriers and facilitators were mapped to behaviour change techniques, focussing on the intervention types of education, persuasion and environmental restructuring, alongside appropriate modes of delivery, and prototype intervention materials were developed. (iii) We iteratively refined these materials in a workshop with multidisciplinary experts and Patient and Public Involvement and Engagement (PPIE) representatives (n = 13) and an (iv) intervention walkthrough with families (n = 5). The design process was informed by the Behaviour Change Wheel, Theoretical framework of acceptability and the Template for intervention description and replication.ResultsSocial influences and motivational factors are likely to influence both uptake and adherence to the intervention. Anticipated emotional reward from participating in research for the benefit of others was indicated to be a strong facilitator for intervention uptake. Alternatives to bathing, having fun with the baby and the night-time routine, alongside family support, were notable facilitators suggested to aid adherence to the intervention. Barriers included hygiene concerns and anticipated negative social appraisal. Barriers and facilitators were mapped to thirty-six behaviour change techniques, focussing on the intervention types of education, persuasion and environmental restructuring, all of which were embedded into the package of support. The prototype intervention materials received positive feedback from the expert workshop and study walkthrough with families. The final package of support comprises printed and digital prompts and cues, a study booklet, video, and digital tool for self-monitoring.ConclusionsThe intervention design process incorporated the ‘real world’ views and experiences of families, experts and PPIE representatives, alongside criteria for designing behavioural interventions. The effectiveness of the package of support will be tested in a feasibility trial and embedded process evaluation.</div

Interview topic guides.

BackgroundBathing babies less frequently and intensively in the first six months of life may prevent eczema, but this has not yet been definitively tested in a randomised controlled trial. Such a trial would require evidence-based support to help parents engage with a minimal bathing routine. The present study reports the development of this support.MethodsWe adopted a four-stage design process: (i) Pregnant women and their families (n = 31) were interviewed to ascertain key barriers and facilitators towards following the minimal bathing intervention. (ii) These barriers and facilitators were mapped to behaviour change techniques, focussing on the intervention types of education, persuasion and environmental restructuring, alongside appropriate modes of delivery, and prototype intervention materials were developed. (iii) We iteratively refined these materials in a workshop with multidisciplinary experts and Patient and Public Involvement and Engagement (PPIE) representatives (n = 13) and an (iv) intervention walkthrough with families (n = 5). The design process was informed by the Behaviour Change Wheel, Theoretical framework of acceptability and the Template for intervention description and replication.ResultsSocial influences and motivational factors are likely to influence both uptake and adherence to the intervention. Anticipated emotional reward from participating in research for the benefit of others was indicated to be a strong facilitator for intervention uptake. Alternatives to bathing, having fun with the baby and the night-time routine, alongside family support, were notable facilitators suggested to aid adherence to the intervention. Barriers included hygiene concerns and anticipated negative social appraisal. Barriers and facilitators were mapped to thirty-six behaviour change techniques, focussing on the intervention types of education, persuasion and environmental restructuring, all of which were embedded into the package of support. The prototype intervention materials received positive feedback from the expert workshop and study walkthrough with families. The final package of support comprises printed and digital prompts and cues, a study booklet, video, and digital tool for self-monitoring.ConclusionsThe intervention design process incorporated the ‘real world’ views and experiences of families, experts and PPIE representatives, alongside criteria for designing behavioural interventions. The effectiveness of the package of support will be tested in a feasibility trial and embedded process evaluation.</div

Participant informed consent forms.

BackgroundBathing babies less frequently and intensively in the first six months of life may prevent eczema, but this has not yet been definitively tested in a randomised controlled trial. Such a trial would require evidence-based support to help parents engage with a minimal bathing routine. The present study reports the development of this support.MethodsWe adopted a four-stage design process: (i) Pregnant women and their families (n = 31) were interviewed to ascertain key barriers and facilitators towards following the minimal bathing intervention. (ii) These barriers and facilitators were mapped to behaviour change techniques, focussing on the intervention types of education, persuasion and environmental restructuring, alongside appropriate modes of delivery, and prototype intervention materials were developed. (iii) We iteratively refined these materials in a workshop with multidisciplinary experts and Patient and Public Involvement and Engagement (PPIE) representatives (n = 13) and an (iv) intervention walkthrough with families (n = 5). The design process was informed by the Behaviour Change Wheel, Theoretical framework of acceptability and the Template for intervention description and replication.ResultsSocial influences and motivational factors are likely to influence both uptake and adherence to the intervention. Anticipated emotional reward from participating in research for the benefit of others was indicated to be a strong facilitator for intervention uptake. Alternatives to bathing, having fun with the baby and the night-time routine, alongside family support, were notable facilitators suggested to aid adherence to the intervention. Barriers included hygiene concerns and anticipated negative social appraisal. Barriers and facilitators were mapped to thirty-six behaviour change techniques, focussing on the intervention types of education, persuasion and environmental restructuring, all of which were embedded into the package of support. The prototype intervention materials received positive feedback from the expert workshop and study walkthrough with families. The final package of support comprises printed and digital prompts and cues, a study booklet, video, and digital tool for self-monitoring.ConclusionsThe intervention design process incorporated the ‘real world’ views and experiences of families, experts and PPIE representatives, alongside criteria for designing behavioural interventions. The effectiveness of the package of support will be tested in a feasibility trial and embedded process evaluation.</div