Focus groups are conventionally run face to face, using a location suitable for researchers and participants. However, to reduce transmission of Covid-19, people were advised to physically distance themselves, to meet outdoors rather than inside and to wear face masks [29], making physical meetings difficult to conduct. Virtual focus groups have been previously reported, with researchers using a variety of techniques such as message boards and video conferencing, with cost of equipment (e.g., webcams) and programmes listed as potential disadvantages [30, 31]. The restrictions imposed to limit the spread of Covid-19 accelerated the widespread adoption of virtual meeting platforms such Zoom for both work and social uses. Indeed, many participants in this study described the benefits of online meetings, indicating that many of the earlier challenges with virtual platforms, such as equipment availability, had been overcome. Table 1 lists potential advantages and disadvantages of physical (under Covid-19 restrictions) and virtual meetings. The assessment was made that, with the ongoing pandemic associated restrictions, the virtual platform was the most appropriate technique to enable the focus groups to be conducted.
Planned changes in pharmacist pre-registration training, including at undergraduate level, will result in newly registered pharmacists registering as independent prescribers [51]. Pharmacist participants expressed concerns about this development, including detraction from training aspects and potential exacerbation of prescribing errors, as previously identified with junior medical staff [52]. The participants placed their views in the context of their own prescribing training, highlighting the struggle that less experienced pharmacists had with the course, and commenting that routine pharmacy work still needed addressing. However, the development is in line with the Carter report and draft workforce strategy, which both envisaged a clinical pharmacy workforce, with pharmacy technicians adopting some of the traditional pharmacist roles [36, 53]. The concerns expressed by pharmacist participants regarding time pressures to complete their tasks suggest that advanced pharmacy technician roles, which would release pharmacist time for prescribing, have still to be adopted.
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Where prescription regimens change frequently, analysis depends on modelling assumptions [111]. This approach has, for example, been used to explore associations between breastfeeding, SES and adult health [112], and between breastfeeding, infant deaths and self-reported ethnic group [113].
To our knowledge, this is the first paper to address the complex questions surrounding incorporation of infant feeding data into healthcare databases used to report patient safety. Other research approaches, such as case series, recruited cohorts and randomised trials, and analytical methods are detailed in guidelines [119], and are outside the scope of this discussion; however, concerns over external generalisation remain [21]. For simplicity, our tentative directed acyclic graphs do not offer comprehensive characterisation of the full range of variables affecting childhood outcomes and breastfeeding, listed above [1, 120, 122, 124, 129].
The inhibitory influence of Noggin4 on Wnt8 signalling was confirmed by its ability to down-regulate the specific β-catenin reporter TOPFlash, which is activated by Wnt8 (Fig. S2f). In contrast, Noggin4 did not antagonize the activity of β-catenin, the intracellular effector of canonical Wnt signalling, in a similar test (Fig. S2f). These findings indicate that Noggin4 may operate at the level of or upstream of Wnt8 receptors.
To act as a natural antagonist of Wnt8, Noggin4 must spread and interact with Wnt8 within the IS in living embryos. To track Noggin4 and Wnt8 diffusion, we produced constructs that encoded secreted fusions of Noggin1/2/4 and Wnt8 with EGFP or TagRFP (Table S1; Fig. S7). Importantly, these fusion proteins retained abilities of their wild-type variants to influence BMP and Wnt signalling reporters (Fig. S8a,b) and to induce, with the collaboration of tBR, secondary heads with cyclopic eyes (Fig. S8c,d).
Animal experiments were performed in accordance with guidelines approved by the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (Moscow, Russia) Animal Committee and handled in accordance with the Animals (Scientific Procedures) Act 1986 and Helsinki Declaration.
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