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In synthetic biology, microbial chassis including yeast Saccharomyces cerevisiae are iteratively engineered with increasing complexity and scale. Wet-lab genetic engineering tools are developed and optimised to facilitate strain construction but are often incompatible with each other due to shared regulatory elements, such as the galactose-inducible (GAL) promoter in S. cerevisiae. Here, we prototyped the cyanamide-induced I−SceI expression, which triggered double-strand DNA breaks (DSBs) for selectable marker removal. We further combined cyanamide-induced I−SceI-mediated DSB and maltose-induced MazF-mediated negative selection for plasmid-free in situ promoter substitution, which simplified the molecular cloning procedure for promoter characterisation. We then characterised three tetracycline-inducible promoters showing differential strength, a non-leaky β-estradiol-inducible promoter, cyanamide-inducible DDI2 promoter, bidirectional MAL32/MAL31 promoters, and five pairs of bidirectional GAL1/GAL10 promoters. Overall, alternative regulatory controls for genome engineering tools can be developed to facilitate genomic engineering for synthetic biology and metabolic engineering applications.

Purpose – Interprofessional practice (IPP) is one way to structure collaborations to more effectively meet the complex needs of students in educational settings. This article explores the lessons learned when one research team implemented interprofessional education (IPE) experiences in partnership with a public elementary school and pre-service professionals from elementary education, special education and communication science and disorders. Design/methodology/approach – This reflective article explores the lived experiences of researchers and partners who completed an IPE experience within one professional development school’s site. Researcher anecdotes are included to support the viewpoints shared. Findings – It was discovered that IPE experiences are essential to facilitate meaningful collaborations for pre-service professionals to learn with and from one another; however, this requires time, preparation and is most effective when teacher mentors and university professors lead with vulnerability and model flexibility. Investment in IPE is challenging but worth the effort when learning outcomes are realized. Originality/value – Specific details regarding the structure of this experience are shared as well as future directional goals for programs hoping to implement IPE in their professional practice programs.

Summary: Background: Large-scale multicentre studies are needed to understand complex relationships between the gut microbiota, health and disease. Interrogating the mucosal microbiota may identify important biology not captured by stool analysis. Gold standard tissue cryopreservation (‘flash freezing’) limits large-scale study feasibility. We aimed to compare gut microbiota in gold standard and pragmatic mucosal biopsy storage conditions. Methods: We collected endoscopic recto-sigmoid biopsies from 20 adults with inflammatory bowel disease. Biopsies were preserved using three methods: (i) flash freezing (most proximal and distal biopsy sites); (ii) nucleic acid preservative reagents (QIAGEN Allprotect®, Invitrogen RNAlater™, and Zymo DNA/RNA Shield™); and (iii) formalin fixation with paraffin embedding (FFPE), which is used to preserve tissue for clinical histopathology within healthcare settings. Microbiota were sequenced on the MiSeq platform (V4 region, 16S rRNA gene). Findings: Tissue microbiota were consistent between most proximal and distal tissue suggesting any within-patient variation observed reflected storage condition, not biopsy location. There was no significant difference in alpha-diversity or microbial community profiles of reagent-preserved versus gold standard tissue. FFPE community structure was significantly dissimilar to other tissue samples, driven by differential relative abundance of obligate gut anaerobes; Faecalibacterium, Anaerostipes and Lachnospiraceae. Despite these differences, tissue microbiota grouped by participant regardless of preservation and storage conditions. Interpretation: Preservative reagents offer a convenient alternative to flash freezing tissue in prospective large-scale mucosal microbiota studies. Whilst less comparable, FFPE provides potential for retrospective microbiota studies using historical samples. Funding: Medical Research Council (MR/T032162/1) and The Leona M. and Harry B. Helmsley Charitable Trust (G-2002-04255).

Background and purpose: The use of magnetic resonance imaging (MRI) for radiotherapy (RT) simulation has grown, prompting quality assurance (QA) guidelines by the Institute of Physics and Engineering in Medicine (IPEM) and the American Association of Physicists in Medicine (AAPM). This study compares a novel multimodality anthropomorphic phantom to an American College of Radiology (ACR) phantom for a subset of these MRI-specific QA tests in RT. Materials and methods: A novel 3D-printed multimodality head-and-neck anthropomorphic phantom was compared to an ACR large MRI phantom. IPEM and AAPM-recommended QA tests were conducted, including informatics/connectivity/data transfer, MRI-CT registration, end-to-end QA, and signal-to-noise ratio (SNR)/percentage integral uniformity (PIU) assessments using RT accessories. Results: Both phantoms were suitable for informatics/connectivity/data transfer. In MRI-CT registration, no errors were found; the ACR phantom offered more quantitative landmarks, while the anthropomorphic phantom provided limited structures. Both phantoms achieved target registration errors (TREs) below 0.97 mm and dice similarity coefficient (DSC) values above 0.9, meeting guidelines. For end-to-end QA, the anthropomorphic phantom facilitated dose measurements of 1.994 Gy versus a calculated 2.01 Gy (−0.8 %). SNR and PIU assessments showed higher values in radiology setups compared to RT setups for both phantoms. Conclusions: Multimodality anthropomorphic phantoms compatible with dosimetric equipment allow realistic end-to-end QA, unlike the ACR phantom. While the ACR phantom is suitable for informatics and MRI-CT registration, anthropomorphic phantoms better represent clinical scenarios. For comprehensive QA, both ACR and anthropomorphic phantoms are required. Additionally, large field-of-view (FOV) phantoms are crucial for evaluating large FOV MRI distortions.

Electronic health records and patient portals are increasingly utilized to enhance research recruitment efficiency, yet response patterns across patient groups remain unclear. We examined 10 studies at Emory Healthcare that used these tools to identify and recruit 24,000 patients over 1 year. Response rates were lower among males and Black individuals, though study interest was higher among respondents. Interest was also greater among those with frequent healthcare interactions and lower comorbidity. In a large academic health system, portal-based recruitment offered a streamlined approach to research recruitment and patient engagement, with minor variations across patient characteristics warranting continued study.

Introduction This analysis aimed to investigate diabetes-specific psychological outcomes among adults with type 1 diabetes (T1D) using hybrid closed-loop (HCL) versus standard therapy.Research design and methods In this multicenter, open-label, randomized, controlled, parallel-group clinical trial, adults with T1D were allocated to 26 weeks of HCL (MiniMed™ 670G) or standard therapy (insulin pump or multiple daily injections without real-time continuous glucose monitoring). Psychological outcomes (awareness and fear of hypoglycemia; and diabetes-specific positive well-being, diabetes distress, diabetes treatment satisfaction, and diabetes-specific quality of life (QoL)) were measured at enrollment, mid-trial and end-trial. Linear mixed models were conducted, using restricted maximum likelihood estimation, unadjusted and adjusted (for covariates: age, sex, diabetes duration, glycated hemoglobin, recent severe hypoglycemia, pre-trial insulin delivery modality, enrollment and mid-study scores).Results 120 participants (mean age 44±12 years) were randomized to intervention (n=61) or standard therapy (n=59). At 13 weeks, the HCL group had better diabetes-specific positive well-being than the standard therapy group (unadjusted: Δ=1.0, p=0.025; adjusted: Δ=1.1, p=0.01), which was maintained at 26 weeks (unadjusted: Δ=0.9, p=0.042; adjusted: Δ=1.0, p=0.023). At 26 weeks, the HCL group also had less diabetes distress (adjusted: Δ=−6.4, p=0.039), fear of hypoglycemia (“maintain high”: adjusted: Δ=−0.8, p=0.034; and “worry”: adjusted: Δ=−1.8, p=0.048), and perceived “unacceptably high glucose levels” (unadjusted: Δ=−1.1, p

Service reservoirs (SRs) are a critical component of drinking water distribution systems that impact water quality. There are no performance measures to quantify or understand this impact. By applying the concept of sink or source behaviour to describe the processes of material accumulation and mobilisation within SRs, this research develops a practical metric to facilitate assessment and quantification of SR performance. It is demonstrated how a few weeks of continuous data from instrumentation deployment at both inlet and outlet of SRs is sufficient to reveal valuable insight into SR and network performance. Through real-world applications, we provide evidence that the metric was able to track SR performance showing both beneficial and detrimental impacts on water quality, as well as quantifying seasonality and the benefits of SR cleaning. Such insight is invaluable for proactive, justifiable, and targeted decisions on the location and frequency of maintenance and management interventions. HIGHLIGHTS A sink or source concept was applied to service reservoirs to describe performance.; A metric was developed to assess and quantify sink and source behaviour in service reservoirs.; Sink behaviour is indicative of good asset performance.; Source behaviour is indicative of deterioration in asset performance.; Service reservoirs can have both a beneficial and negative impact on water quality.;

Surface water flooding (SWF) is a severe hazard associated with extreme convective rainfall, whose spatial and temporal sparsity belie the significant impacts it has on populations and infrastructure. Forecasting the intense convective rainfall that causes most SWF on the temporal and spatial scales required for effective flood forecasting remains extremely challenging. National-scale flood forecasts are currently issued for the UK and are well regarded amongst flood responders, but there is a need for complementary enhanced regional information. Here we present a novel SWF-forecasting method, FOREWARNS (Flood fOREcasts for Surface WAter at a RegioNal Scale), that aims to fill this gap in forecast provision. FOREWARNS compares reasonable worst-case rainfall from a neighbourhood-processed, convection-permitting ensemble forecast system against pre-simulated flood scenarios, issuing a categorical forecast of SWF severity. We report findings from a workshop structured around three historical flood events in Northern England, in which forecast users indicated they found the forecasts helpful and would use FOREWARNS to complement national guidance for action planning in advance of anticipated events. We also present results from objective verification of forecasts for 82 recorded flood events in Northern England from 2013–2022, as well as 725 daily forecasts spanning 2019–2022, using a combination of flood records and precipitation proxies. We demonstrate that FOREWARNS offers good skill in forecasting SWF risk, with high spatial hit rates and low temporal false alarm rates, confirming that user confidence is justified and that FOREWARNS would be suitable for meeting the user requirements of an enhanced operational forecast.