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Ascorbic acid (AA, vitamin C) and dehydroascorbic acid (DHA) constitute a biological couple. No technique can accurately, independently, and simultaneously quantify both members of the couple in animal and human samples, thereby constraining advances in physiology and pathophysiology. Here we describe a new UPLC/MS/MS method to measure both compounds directly and independently in human plasma. Lower limits of quantification were 16 nM, with linear coefficients >0.99 over a 100-fold concentration range. The method was stable and reproducible with

Temperature-related mortality mostly affects older people and is attributable to a combination of factors. We focussed on a key non-temperature factor – rising longevity – and aimed to quantify its reciprocal relation with temperature-related mortality risk in Spain over 1980–2018.We obtained average annual temperature-attributable deaths among people aged 65y+, by sex and age group, for different temperature ranges (extreme cold, moderate cold, moderate heat, and extreme heat), from a previous study. Combining this with population and mortality data as well as life table information, we used: (i) a counterfactual approach to assess the contribution of rising longevity to changes in the absolute risk of temperature-related mortality, and (ii) decomposition to assess the contribution of changes in temperature-related mortality to changes in longevity and its variation (lifespan inequality).Rising longevity led to considerable declines in the absolute risk of temperature-related mortality in females and males across the entire temperature range. For extreme heat, it accounted for about a 30% decrease in absolute risk (half of the total decrease over the study period). For moderate and extreme cold, it accounted for about a 20% fall in absolute risk (a quarter of the total fall). In the opposite direction, changing patterns of temperature-related deaths contributed to higher life expectancy (accounting for > 20% of the total rise in both females and males) but also higher lifespan inequality amongst older people. Most of the influence (about 80%) was via moderate cold, but declines in risk at both moderate and extreme heat led to small rises in life expectancy.Our study points to the benefits of adopting risk-reduction strategies that aim, not only at modifying hazards and reducing exposure, but that also address socially-generated vulnerability among older people. This includes ensuring that lifespans lengthen primarily through increases in years lived in good health.

Abstract Background The clinical benefit of coronavirus disease 2019 (COVID-19) treatments against new circulating variants remains unclear. We sought to describe characteristics and clinical outcomes of highest risk patients with COVID-19 receiving early COVID-19 treatments in Scotland. Methods Retrospective cohort study of non-hospitalized patients diagnosed with COVID-19 from December 1, 2021–October 25, 2022, using Scottish administrative health data. We included adult patients who met ≥ 1 of the National Health Service highest risk criteria for early COVID-19 treatment and received outpatient treatment with sotrovimab, nirmatrelvir/ritonavir or molnupiravir, or no early COVID-19 treatment. Index date was defined as the earliest of COVID-19 diagnosis or early COVID-19 treatment. Baseline characteristics and acute clinical outcomes in the 28 days following index were reported. Values of ≤ 5 were suppressed. Results In total, 2548 patients were included (492: sotrovimab, 276: nirmatrelvir/ritonavir, 71: molnupiravir, and 1709: eligible highest risk untreated). Patients aged ≥ 75 years accounted for 6.9% (n = 34/492), 21.0% (n = 58/276), 16.9% (n = 12/71) and 13.2% (n = 225/1709) of the cohorts, respectively. Advanced renal disease was reported in 6.7% (n = 33/492) of sotrovimab-treated and 4.7% (n = 81/1709) of untreated patients, and ≤ 5 nirmatrelvir/ritonavir-treated and molnupiravir-treated patients. All-cause hospitalizations were experienced by 5.3% (n = 25/476) of sotrovimab-treated patients, 6.9% (n = 12/175) of nirmatrelvir/ritonavir-treated patients, ≤ 5 (suppressed number) molnupiravir-treated patients and 13.3% (n = 216/1622) of untreated patients. There were no deaths in the treated cohorts; mortality was 4.3% (n = 70/1622) among untreated patients. Conclusions Sotrovimab was often used by patients who were aged

Abstract Vivid structural colours in butterflies are caused by photonic nanostructures scattering light. Structural colours evolved for numerous biological signalling functions and have important technological applications. Optically, such structures are well understood, however insight into their development in vivo remains scarce. We show that actin is intimately involved in structural colour formation in butterfly wing scales. Using comparisons between iridescent (structurally coloured) and non-iridescent scales in adult and developing H. sara, we show that iridescent scales have more densely packed actin bundles leading to an increased density of reflective ridges. Super-resolution microscopy across three distantly related butterfly species reveals that actin is repeatedly re-arranged during scale development and crucially when the optical nanostructures are forming. Furthermore, actin perturbation experiments at these later developmental stages resulted in near total loss of structural colour in H. sara. Overall, this shows that actin plays a vital and direct templating role during structural colour formation in butterfly scales, providing ridge patterning mechanisms that are likely universal across lepidoptera.

Summary: Phelan-McDermid syndrome (PMS) is a neurodevelopmental disorder caused by haploinsufficiency of the SHANK3 gene. Approximately 25% of individuals with PMS have epilepsy. Treatment of epilepsy in PMS may require multiple anticonvulsants, and in a minority of cases, seizures remain poorly controlled. Converging lines of evidence in different experimental models indicate that the Ras-ERK pathway is implicated in the pathophysiology of seizure generation and neurobehavioral symptoms in PMS. The goal of this study was to evaluate the safety, tolerability, and efficacy in treating seizures in adults and adolescents with PMS using AMO-01, a Ras-ERK pathway inhibitor. A single 6-hour intravenous infusion of AMO-01 at 120 mg/m2 was administered to six participants using an open-label design. Safety was assessed during the infusion and for 4 weeks post-infusion. Caregivers completed seizure diaries and recorded individual seizures during a baseline period and for 4 weeks following the infusion. Exploratory clinical and biomarker assessments were completed throughout the study. AMO-01 was well tolerated, with no serious adverse events (AEs) reported. All AEs were mild or moderate in severity. Seizures were reduced by at least 25% compared to baseline at each follow-up (weeks 1, 2, and 4). Exploratory clinical measures did not change significantly from baseline, but visual evoked potentials (VEPs) and phosphorylated ERK blood levels revealed trending changes in a subset of participants. These results provide preliminary support for the safety of AMO-01 and its efficacy in reducing seizures in adults with PMS. Future placebo-controlled studies with larger sample sizes and repeated dosing are warranted.

Introduction Current dietary assessment methods face challenges in accurately capturing individuals’ dietary habits, undermining the efficacy of public health strategies. The ‘Standardised and Objective Dietary Intake Assessment Tool’ (SODIAT)-1 study aims to assess the effectiveness of three emerging technologies (urine and capillary blood biomarkers, and wearable camera technology) and two online self-reporting dietary assessment tools to monitor dietary intake. Methods This randomised controlled crossover trial will recruit 30 participants (aged 18-70 years and BMI of 20-30 kg/m 2) from Imperial College London and the University of Reading. Exclusion criteria include recent weight change, food allergies/intolerances, following restrictive diets, certain health conditions and medication use. Interested volunteers will be directed to an online screening questionnaire via REDCap and eligible participants will attend a pre-study visit. Volunteers will consume, in a random order, two highly-controlled diets (compliant and non-compliant with UK guidelines) for four days each. Each study arm will be separated by at least one-week. During each test period, dietary intake will be monitored continuously using wearable cameras and self-recorded using eNutri (food frequency questionnaire) and Intake24 (24-hour dietary recall). Urine and capillary blood samples will be collected for biomarker analysis. Data analysis will assess the accuracy of dietary reporting across these methods using Lin’s concordance correlation coefficient. Discussion and ethical considerations This study introduces a novel approach to dietary assessment, addressing the limitations of traditional methods by reducing misreporting and enhancing inclusivity, particularly for underrepresented populations with literacy or language barriers. However, challenges persist, such as variability in biomarker data due to failure to adhere to sample storage requirements and the practicalities of continuously wearing cameras. To protect privacy, participants will be instructed to remove cameras at inappropriate times, and artificial intelligence will be used to blur all images captured apart from food.

ABSTRACT Microorganisms with simplified genomes represent interesting cell chassis for systems and synthetic biology. However, genome reduction can lead to undesired traits, such as decreased growth rate and metabolic imbalances. To investigate the impact of genome reduction on Escherichia coli strain DGF-298, a strain in which ~ 36% of the genome has been removed, we reconstructed a strain-specific metabolic model (iAC1061), investigated the regulation of gene expression using iModulon-based transcriptome analysis, and performed adaptive laboratory evolution to let the strain correct potential imbalances that arose during its simplification. The model notably predicted that the removal of all three key pathways for glycolaldehyde disposal in this microorganism would lead to a metabolic bottleneck through folate starvation. Glycolaldehyde is also known to cause self-generation of reactive oxygen species, as evidenced by the increased expression of oxidative stress resistance genes in the SoxS iModulon. The reintroduction of the aldA gene, responsible for one native glycolaldehyde disposal route, alleviated the constitutive oxidative stress response. Our results suggest that systems-level approaches and adaptive laboratory evolution have additive benefits when trying to repair and optimize genome-engineered strains.IMPORTANCEGenomic streamlining can be employed in model organisms to reduce complexity and enhance strain predictability. One of the most striking examples is the bacterial strain Escherichia coli DGF-298, notable for having over one-third of its genome deleted. However, such extensive genome modifications raise the question of how similar this simplified cell remains when compared with its parent, and what are the possible unintended consequences of this simplification. In this study, we used metabolic modeling along with iModulon-based transcriptomic analysis in different growth conditions to assess the impact of genome reduction on metabolism and gene regulation. We observed little impact of genomic reduction on the regulatory network of E. coli DGF-298 and identified a potential metabolic bottleneck leading to the constitutive activity of the SoxS iModulon. We then leveraged the model's predictions to successfully restore SoxS activity to the basal level.

Background: Extensive evidence links both cold and hot temperatures to an increased incidence of occupational injuries. Contextual modifiers of the temperature-injury association have been scarcely researched. The present study addresses temporal and spatial variations to identify factors associated with (mal)adaptation to heat and cold among Spanish workers. Methods: We assessed the association between daily mean temperature and work injuries using quasi-Poisson time-series regression models in 48 Spanish provinces over the period 1988–2019, with comparative analyses with census and economic data for the sub-periods 1989–1993, 1999–2003, 2009–2013 and 2015–2019. We explored the spatial and spatiotemporal modification of the association by demographic and socioeconomic variables via cross-sectional and longitudinal meta-regressions. Findings:: We found an increased risk of work-injuries by 4 % [95 % CI: 3 %-6 %] and 12 % [95 % CI: 10 %-13 %], for the 1st and 99th percentiles of temperature, respectively, for period 1988–2019. Heat had a greater overall impact than cold, and the groups more vulnerable to heat were male workers, under 35 years, and working in agriculture, construction and hostelry. Vulnerability to heat was highest in the earliest sub-period, while vulnerability to cold rose during periods of both economic expansion and recession. High educational attainment emerged as a protective factor during the warm months in the cross-sectional meta-regressions. Conclusions: Our findings suggest an adaptation of Spanish workers to high temperatures over time. However, preventive measures are needed for traditionally exposed workers (agriculture and construction), non-traditionally vulnerable sectors (hostelry), and young, male, and less educated workers during warm months. For cold vulnerability, targeted measures should focus on women, the elderly, and tertiary service workers, especially in colder regions. Addressing temperature vulnerability would enhance worker safety, reduce injuries, and yield economic benefits.

Abstract Many of the proposed solutions to the global biodiversity crisis rely on national governments to act. The conservation movement needs to motivate governments or face an ongoing extinction crisis. Here we explore how linking biodiversity to electoral systems may assist in motivating government action. Using Australia as a case study, we analyze the intersection of 151 electoral districts and 1651 threatened species. We show all districts contain at least 14 threatened species. Half of the species analyzed (n = 801, 49%) are confined to one district (n = 44), with 1345 (81%) species intersecting with less than five. This geographical information shows that alongside local social and economic issues, the threatened species crisis can be made relevant to all Australian elected representatives. Locally relevant information can encourage integration of species needs into the scope of political representation. As such, linking biodiversity to political geography offers a potential pathway to creating transformative change.

Population ageing is expected to lead to significant rises in climate risks because vulnerability rises sharply throughout people’s later years. When assessing the vulnerability of older people, however, what’s important isn’t the number of years someone has lived (i.e. “chronological age”) but rather their functional abilities and characteristics; the latter is better captured by remaining life expectancy or “prospective age”. Here, we show that assessing growth in the size of older populations using a prospective rather than chronological age perspective can help avoid overestimates of future risks to climate change. Compared to an analysis based on chronological age, the projected increase in the vulnerable population share seen in the prospective age analysis is considerably lower. The differences between the two perspectives increase with age, decrease with country income level, and are larger in futures that give priority to sustainable development. Thus, while ageing certainly poses major challenges to societies facing climate change, these may be smaller than thought. Prospective age offers a relatively easily implemented alternative for projecting future vulnerability that better accounts for rising longevity.

Abstract Background The impact of the constantly evolving severe acute respiratory syndrome coronavirus 2 on the effectiveness of early coronavirus disease 2019 (COVID-19) treatments is unclear. Here, we report characteristics and acute clinical outcomes of patients with COVID-19 treated with a monoclonal antibody (mAb; presumed to be sotrovimab) across six distinct periods covering the emergence and predominance of Omicron subvariants (BA.1, BA.2, and BA.5) in England. Methods Retrospective cohort study using data from the Hospital Episode Statistics database from January 1–July 31, 2022. Included patients received a mAb delivered by a National Health Service (NHS) hospital as a day-case, for which the primary diagnosis was COVID-19. Patients were presumed to have received sotrovimab based on NHS data showing that 99.98% of COVID-19-mAb-treated individuals received sotrovimab during the study period. COVID-19-attributable hospitalizations were reported overall and across six distinct periods of Omicron subvariant prevalence. Subgroup analyses were conducted in patients with severe renal disease and active cancer. Results Among a total of 10,096 patients, 1.0% (n = 96) had a COVID-19-attributable hospitalization, 4.6% (n = 465) had a hospital visit due to any cause, and 0.3% (n = 27) died due to any cause during the acute period. COVID-19-attributable hospitalization rates were consistent among subgroups, and no significant differences were observed across periods of Omicron subvariant predominance. Conclusions Levels of COVID-19-attributable hospitalizations and deaths were low in mAb-treated patients and among subgroups. Similar hospitalization rates were observed whilst Omicron BA.1, BA.2, and BA.5 were predominant, despite reported reductions in in vitro neutralization activity of sotrovimab against BA.2 and BA.5.

Between July 2020 and January 2021, 230 principal caregivers completed a questionnaire to measure proxy-assessed health-related quality of life outcomes (HRQoL), behavioral outcomes in children with syndromic autism spectrum disorders and COVID-19 induced changes to lifestyle and environments. HRQoL and behavioral outcomes reported earlier during the pandemic were generally worse compared to those reported later. COVID-19 induced reduction to a caregiver's mental health appointments, and hours spent watching TV were associated with decreases in HRQoL and increased the likelihood of problematic behaviors. Increasing time outdoors and time away from digital devices were positively associated with HRQoL and behaviors and might protect children from COVID-19 induced restrictions.

Objective: To search for, review and synthesise studies of the effectiveness and cost-effectiveness of weight management schemes for the under fives. Data sources: MEDLINE [Ovid], MEDLINE In-Process [Ovid], EMBASE [Ovid], CAB [Ovid], Health Management Information Consortium [Ovid], The Cochrane Database of Systematic Reviews, Cochrane Register of Controlled Trials, Science Citation Index Expanded [Web of Science], Conference Proceedings Citation Index [The Web of Science], Database of Abstract Reviews [CRD; Centre for Reviews and Dissemination], HTA [CRD], PsycINFO [Ebsco], NHS CRD. These databases were searched from 1990 to February 2009. Supplementary internet searches were additionally conducted. Review methods: Relevant clinical effectiveness studies were identified in two stages. Titles and abstracts returned by the search strategy were examined independently by three researchers and screened for possible inclusion. Disagreements were resolved by discussion. Full texts of the identified studies were obtained. Three researchers examined these independently for inclusion or exclusion, and disagreements were again resolved by discussion. Results: One of the randomised controlled trials (RCTs) was from the UK. It measured the effects of a physical activity intervention for children in nurseries combined with home-based health education for their parents; this was compared to usual care. The main outcome measure was body mass index (BMI); secondary measures were weight and physical activity. At the 12-month follow-up, no statistically significant differences were found between the groups on any measure. However, a trend, favouring the intervention, was found for BMI and weight. The other two RCTs were from the USA. The larger trial investigated the effects of a combined preschool and home intervention in African American and Latino communities. Nutrition education and physical activity programmes were aimed at under fives in preschool. The home component consisted of related health education and homework for the parents, who received a small financial reward on completion. The 1- and 2-year results for the African American sites showed a significantly slower rate of increase in BMI than for results at baseline, for the intervention group than for the control group. However, in the Latino communities no such differences were found. The second US trial was a much smaller home-based parental education programme in Native American communities in the USA and Canada. The intervention consisted of a parental skills course for parents to improve their children’s diet and physical activity. This was compared with a course providing skills to improve child behaviour. Follow-up was at 16 weeks and showed no significant differences between groups in BMI. Conclusions: No controlled trials addressing the issue of treating obesity or evidence of cost-effectiveness studies in the under fives’ population were found. From the three prevention studies, apart from the larger US trial, the interventions showed no statistically significant differences in BMI and weight between the intervention and control groups (although there was some evidence of positive trends for BMI and weight). It should also be noted that these conclusions are based on only three dissimilar studies, thereby making the drawing of firm conclusions difficult. Further research is urgently needed in well-designed UK-based RCTs of weight management schemes aimed at the prevention of obesity, that combine with cost-effectiveness studies targeted at preschool children with long-term follow-up.

Spin-split heavy-hole gases in strained germanium quantum wells were characterized by polarisation-resolved terahertz time-domain spectroscopy. Effective masses, carrier densities, g-factors, transport lifetimes, mobilities and Rashba spin-splitting energies were evaluated, giving quantitative insights into the influence of strain. The Rashba coefficient was found to lower for samples with higher biaxial compressive strain, while heavy-hole mobilities were enhanced to over $1.5\times {10}^{6}$ cm ^2 V ^−1 s ^−1 at 3 K. This high mobility enabled the observation of the optical quantum Hall effect at terahertz frequencies for spin-split two-dimensional heavy-holes, evidenced as plateaux in the transverse magnetoconductivity at even and odd filling factors.

Background We assessed the effectiveness of sotrovimab vs no early COVID-19 treatment in highest-risk COVID-19 patients during Omicron predominance.Methods Retrospective cohort study using the Discover dataset in North West London. Included patients were non-hospitalised, aged ≥12 years and met ≥1 National Health Service highest-risk criterion for sotrovimab treatment. We used Cox proportional hazards models to compare HRs of 28-day COVID-19-related hospitalisation/death between highest-risk sotrovimab-treated and untreated patients. Age, renal disease and Omicron subvariant subgroup analyses were performed.Results We included 599 sotrovimab-treated patients and 5191 untreated patients. Compared with untreated patients, the risk of COVID-19 hospitalisation/death (HR 0.50, 95% CI 0.24, 1.06; p=0.07) and the risk of COVID-19 hospitalisation (HR 0.43, 95% CI 0.18, 1.00; p=0.051) were both lower in the sotrovimab-treated group; however, statistical significance was not reached. In the ≥65 years and renal disease subgroups, sotrovimab was associated with a significantly reduced risk of COVID-19 hospitalisation, by 89% (HR 0.11, 95% CI 0.02, 0.82; p=0.03) and 82% (HR 0.18, 95% CI 0.05, 0.62; p=0.007), respectively.Conclusions Risk of COVID-19 hospitalisation in sotrovimab-treated patients aged ≥65 years and with renal disease was significantly lower compared with untreated patients. Overall, risk of hospitalisation was also lower for sotrovimab-treated patients, but statistical significance was not reached.

Background Health impact assessment (HIA) is a widely used process that aims to identify the health impacts, positive or negative, of a policy or intervention that is not necessarily placed in the health sector. Most HIAs are done prospectively and aim to forecast expected health impacts under assumed policy implementation. HIAs may quantitatively and/or qualitatively assess health impacts, with this study focusing on the former. A variety of quantitative modelling methods exist that are used for forecasting health impacts, however, they differ in application area, data requirements, assumptions, risk modelling, complexities, limitations, strengths, and comprehensibility. We reviewed relevant models, so as to provide public health researchers with considerations for HIA model choice.Methods Based on an HIA expert consultation, combined with a narrative literature review, we identified the most relevant models that can be used for health impact forecasting. We narratively and comparatively reviewed the models, according to their fields of application, their configuration and purposes, counterfactual scenarios, underlying assumptions, health risk modelling, limitations and strengths.Results Seven relevant models for health impacts forecasting were identified, consisting of (i) comparative risk assessment (CRA), (ii) time series analysis (TSA), (iii) compartmental models (CMs), (iv) structural models (SMs), (v) agentbased models (ABMs), (vi) microsimulations (MS), and (vii) artificial intelligence (AI)/machine learning (ML). These models represent a variety in approaches and vary in the fields of HIA application, complexity and comprehensibility. We provide a set of criteria for HIA model choice. Researchers must consider that model input assumptions match the available data and parameter structures, the available resources, and that model outputs match the research question, meet expectations and are comprehensible to end-users.Conclusion The reviewed models have specific characteristics, related to available data and parameter structures, computational implementation, interpretation and comprehensibility, which the researcher should critically consider before HIA model choice.

Objective: this article aims to answer the following questions: 1. Can generative artificial intelligence be a subject of copyright law? 2. What risks the unregulated use of generative artificial intelligence systems can cause? 3. What legal gaps should be filled in to minimize such risks?Methods: comparative legal analysis, sociological method, concrete sociological method, quantitative data analysis, qualitative data analysis, statistical analysis, case study, induction, deduction.Results: the authors identified several risks of the unregulated usage of generative artificial intelligence in the creative industry, among which are: violation of copyright and labor law, violation of consumers rights and the rise of public distrust in government. They suggest that a prompt development of new legal norms can minimize these risks. In conclusion, the article constants that states have already begun to realize that the negative impact of generative artificial intelligence on the creative industry must not be ignored, hence the development of similar legal regulations in states with completely different regimes.Scientific novelty: the article provides a comprehensive study of the impact of generative artificial intelligence on the creative industry from two perspectives: the perspective of law and the perspective of the industry. The empirical basis of it consists of two international surveys and an expert opinion of a representative of the industry. This approach allowed the authors to improve the objectivity of their research and to obtain results that can be used for finding a practical solution for the identified risks. The problem of the ongoing development and popularization of generative artificial intelligence systems goes beyond the question “who is the author?” therefore, it needs to be solved by introduction of other than the already existing mechanisms and regulations - this point of view is supported not only by the results of the surveys but also by the analysis of current lawsuits against developers of generative artificial intelligence systems.Practical significance: the obtained results can be used to fasten the development of universal legal rules, regulations, instruments and standards, the current lack of which poses a threat not only to human rights, but also to several sectors within the creative industry and beyond.

Physical literacy (PL), a concept commonly associated with the early years, physical education, and youth sport development, can become a meaningful determinant of health and longevity for the adult and older adult population. A review of 55 recent publications from 2018 to 2023 that encompassed physical literacy conceptual frameworks, assessments, and intervention-based studies was undertaken through an heuristic inspired by the philosophy which gave birth to PL. With particular interest in how PL has evolved in response to the needs of an aging population, this position paper tracks a key shift in focus from the individual to the relational context. It references positive interaction and social participation in recent models as significant features of an across-the-lifespan PL perspective.The concluding position is that fostering joyful inter-action be at the heart of PL promotion, resource development and assessment practices, especially in the case of an aging population.

ABSTRACT Many pathogenic Gram-negative bacteria use repeats-in-toxin adhesins for colonization and biofilm formation. In the cholera agent Vibrio cholerae, flagellar-regulated hemagglutinin A (FrhA) enables these functions. Using bioinformatic analysis, a sugar-binding domain was identified in FrhA adjacent to a domain of unknown function. AlphaFold2 indicated the boundaries of both domains to be slightly shorter than previously predicted and assisted in the recognition of the unknown domain as a split immunoglobulin-like fold that can assist in projecting the sugar-binding domain toward its target. The AlphaFold2-predicted structure is in excellent agreement with the molecular envelope obtained from small-angle X-ray scattering analysis of a recombinant construct spanning the sugar-binding and unknown domains. This two-domain construct was probed by glycan micro-array screening and showed binding to mammalian fucosylated glycans, some of which are characteristic erythrocyte markers and intestinal cell epitopes. Isothermal titration calorimetry further showed the construct-bound l-fucose with a Kd of 21 µM. Strikingly, this recombinant protein construct bound and lysed erythrocytes in a concentration-dependent manner, and its hemolytic activity was blocked by the addition of l-fucose. A protein ortholog construct from Aeromonas veronii was also produced and showed a similar glycan-binding pattern, binding affinity, erythrocyte-binding, and hemolytic activities. As demonstrated here with Hep-2 cells, fucose-based inhibitors of this sugar-binding domain can potentially be developed to block colonization by V. cholerae and other pathogenic bacteria that share this adhesin domain.IMPORTANCEThe bacterium, Vibrio cholerae, which causes cholera, uses an adhesion protein to stick to human cells and begin the infection process. One part of this adhesin protein binds to a particular sugar, fucose, on the surface of the target cells. This binding can lead to colonization and killing of the cells by the bacteria. Adding l-fucose to the bacteria before they bind to the human cells can prevent attachment and has promise as a preventative drug to protect against cholera.

Children with autism have a significantly lower quality of life compared with their neurotypical peers. While multiple studies have quantified the impact of autism on health-related quality of life (HRQoL) through standardized surveys such as the PedsQL, none have specifically investigated the impact of syndromic autism. Here we evaluate HRQoL in children diagnosed with three genetic disorders that strongly predispose to syndromic autism: Phelan-McDermid syndrome (PMD), Rett syndrome (RTT), and SYNGAP1-related intellectual disability (SYNGAP1-ID). We find the most severely impacted dimension is physical functioning. Strikingly, syndromic autism results in worse quality of life than other chronic disorders including idiopathic autism. This study demonstrates the utility of caregiver surveys in prioritizing phenotypes, which may be targeted as clinical endpoints for genetically defined ASDs.

Background: This study estimated utility values for non-alcoholic steatohepatitis (NASH). Previous studies have assumed that health-related quality of life does not vary between the early stages of NASH. Materials &Methods: Discrete choice experiment (DCE) surveys estimated the value of avoiding fibrosis progression. Patients also completed the EQ-5D-5L. Marginal rates of substitution estimated utility change associated with fibrosis progression. Results: DCE surveys were completed by the UK general public (n = 520) and patients with NASH (n = 154). The utility decline between fibrosis stages F1 and F4 decompensated was between -0.521 to -0.646 (depending on method). Conclusion: Three methods were used to estimate utilities for NASH, each one showed sensitivity to advancing fibrosis, including in the early stages, which is often considered asymptomatic.

Background: A number of studies have reported reductions in mortality risk due to heat and cold over time. However, questions remain about the drivers of these adaptation processes to ambient temperatures. We aimed to analyse the demographic and socioeconomic drivers of the downward trends in vulnerability to heat- and cold-related mortality observed in Spain during recent decades (1980–2018). Methods: We collected data on all-cause mortality, temperature and relevant contextual indicators for 48 provinces in mainland Spain and the Balearic Islands between Jan 1, 1980, and Dec 31, 2018. Fourteen contextual indicators were analysed representing ageing, isolation, urbanicity, heating, air conditioning (AC), house antiquity and ownership, education, life expectancy, macroeconomics, socioeconomics, and health investment. The statistical analysis was separately performed for the range of months mostly causing heat- (June-September) and cold- (October-May) related mortality. We first applied a quasi-Poisson generalised linear regression in combination with distributed lag non-linear models (DLNM) to estimate province-specific temperature-mortality associations for different periods, and then we fitted univariable and multivariable multilevel spatiotemporal meta-regression models to evaluate the effect modification of the contextual characteristics on heat- and cold-related mortality risks over time. Findings: The average annual mean temperature has risen at an average rate of 0·36 °C per decade in Spain over 1980–2012, although the increase in temperature has been more pronounced in summer (0·40 °C per decade in June-September) than during the rest of the year (0·33 °C per decade). This warming has been observed, however, in parallel with a progressive reduction in the mortality risk associated to both hot and cold temperatures. We found independent associations for AC with heat-related mortality, and heating with cold-related mortality. AC was responsible for about 28·6% (31·5%) of the decrease in deaths due to heat (extreme heat) between 1989 and 1993 and 2009–2013, and heating for about 38·3% (50·8%) of the reductions in deaths due to cold (extreme cold) temperatures. Ageing (ie, proportion of population over 64 years) attenuated the decrease in cold-related mortality. Interpretation: AC and heating are effective societal adaptive measures to heat and cold temperatures. This evidence holds important implications for climate change health adaptation policies, and for the projections of climate change impacts on human health.

IntroductionDevelopmental synaptopathies are neurodevelopmental disorders caused by genetic mutations disrupting the development and function of neuronal synapses.MethodsWe administered the validated Social Responsiveness Scale, Second Edition (SRS-2) to investigate the phenotypic presentation of social-behavioral impairments for the developmental synaptopathy—SYNGAP1-related Intellectual Disability (SYNGAP1-ID) (n = 32) compared with a phenotypically similar disorder Phelan-McDermid syndrome (PMD) (n = 27) and healthy controls (n = 43). A short form SRS-2 analysis (n = 85) was also conducted.ResultsBoth SYNGAP1-ID and PMD had significantly elevated total and subcategory T-scores, with no significant score differences between SYNGAP1-ID and PMD, consistent between the full and short form. Mild to severe deficiencies in reciprocal social behavior were found in 100% of PMD individuals and 87.1% of SYNGAP1-ID individuals. Surprisingly, a positive correlation between age and total score was discovered for SYNGAP1-ID participants and not found in individuals with PMD or healthy controls.DiscussionThe short form demonstrated greater utility for SYNGAP1-ID participants due to lower item-omission rates. In conclusion, significant impairment in reciprocal social behaviors is highly prevalent in SYNGAP1-ID.

In silico genome mining provides easy access to secondary metabolite biosynthetic gene clusters (BGCs) encoding the biosynthesis of many bioactive compounds, which are the basis for many important drugs used in human medicine. However, the association between BGCs and other functions encoded in the genomes of producers have remained elusive. Here, we present a systems biology workflow that integrates genome mining with a detailed pangenome analysis for detecting genes associated with a particular BGC. We analyzed 3,889 enterobacterial genomes and found 13,266 BGCs, represented by 252 distinct BGC families and 347 additional singletons. A pangenome analysis revealed 88 genes putatively associated with a specific BGC coding for the colon cancer-related colibactin that code for diverse metabolic and regulatory functions. The presented workflow opens up the possibility to discover novel secondary metabolites, better understand their physiological roles, and provides a guide to identify and analyze BGC associated gene sets.

**Background:** Late-onset Pompe disease (LOPD) is a rare, progressive neuromuscular condition typically characterized by weakness of skeletal muscles, including those involved in respiration and diaphragmatic dysfunction. Individuals with LOPD typically eventually require mobility and/or ventilatory support. **Objectives:** This study aimed to develop health state vignettes and estimate health state utility values for LOPD in the United Kingdom. **Methods:** Vignettes were developed for 7 health states of LOPD with states defined in terms of mobility and/or ventilatory support. Vignettes were drafted based on patient-reported outcome data from the Phase 3 PROPEL trial (NCT03729362) and supplemented by a literature review. Qualitative interviews with individuals living with LOPD and clinical experts were conducted to explore the health-related quality-of-life (HRQoL) impact of LOPD and to review the draft vignettes. Vignettes were finalized following a second round of interviews with individuals living with LOPD and used in health state valuation exercises with people of the UK population. Participants rated the health states using the EQ-5D-5L, visual analogue scale, and time trade-off interviews. **Results:** Twelve individuals living with LOPD and 2 clinical experts were interviewed. Following the interviews, 4 new statements were added regarding dependence on others, bladder control problems, balance issues/fear of falling, and frustration. One hundred interviews with a representative UK population sample were completed. Mean time trade-off utilities ranged from 0.754 (SD = 0.31) (no support) to 0.132 (SD = 0.50) (invasive ventilatory and mobility support–dependent). Similarly, EQ-5D-5L utilities ranged from 0.608 (SD = 0.12) to -0.078 (SD = 0.22). **Discussion:** The utilities obtained in the study are consistent with utilities reported in the literature (0.670-0.853 for nonsupport state). The vignette content was based on robust quantitative and qualitative evidence and captured the main HRQoL impacts of LOPD. The general public rated the health states consistently lower with increasing disease progression. There was greater uncertainty around utility estimates for the severe states, suggesting that participants found it harder to rate them. **Conclusion:** This study provides utility estimates for LOPD that can be used in economic modeling of treatments for LOPD. Our findings highlight the high disease burden of LOPD and reinforce the societal value of slowing disease progression.

Abstract Background A novel treatment has been developed for erythropoietic protoporphyria (EPP) (a rare condition that leaves patients highly sensitive to light). To fully understand the burden of EPP and the benefit of treatment, a novel patient reported outcome (PRO) measure was developed called the EPP-QoL. This report describes work to support the validation of this measure. Methods Secondary analysis of trial data was undertaken. These analyses explored the underlying factor structure of the measure. This supported the deletion of some items. Further work then explored the reliability of these factors, their construct validity and estimates of meaningful change. Results The factor analyses indicated that the items could be summarised in terms of two factors. One of these was labelled EPP Symptoms and the other EPP Wellbeing, based on the items included in the domain. EPP Symptoms had evidence to support its reliability and validity. EPP Wellbeing had poor psychometric properties. Conclusions Based on the analysis it was recommended to drop the EPP Wellbeing domain (and associated items). EPP Symptoms, despite limitations in the development of items, showed evidence of validity. This work is consistent with the recommendations of a task force that provided recommendations regarding the development, modification and use of PROs in rare diseases.

Abstract Mesoplasma florum, a fast‐growing near‐minimal organism, is a compelling model to explore rational genome designs. Using sequence and structural homology, the set of metabolic functions its genome encodes was identified, allowing the reconstruction of a metabolic network representing ˜ 30% of its protein‐coding genes. Growth medium simplification enabled substrate uptake and product secretion rate quantification which, along with experimental biomass composition, were integrated as species‐specific constraints to produce the functional iJL208 genome‐scale model (GEM) of metabolism. Genome‐wide expression and essentiality datasets as well as growth data on various carbohydrates were used to validate and refine iJL208. Discrepancies between model predictions and observations were mechanistically explained using protein structures and network analysis. iJL208 was also used to propose an in silico reduced genome. Comparing this prediction to the minimal cell JCVI‐syn3.0 and its parent JCVI‐syn1.0 revealed key features of a minimal gene set. iJL208 is a stepping‐stone toward model‐driven whole‐genome engineering.

Researchers established that parental vaccination status often predicts that of their children, but a limited number of studies have examined factors influencing dyadic concordance or discordance (i.e., same or different vaccination status or intent for both members). We investigated how child versus parent age as well as parents’ perceptions of their respective friends’ immunization behavior impacted un/vaccinated parents’ decisions regarding vaccinating their child. An online survey obtained the COVID-19 vaccination status and views of 762 parents of 5–17-year-old children. More than three-quarters of all dyads were concordant; 24.1% of vaccinated parents would not vaccinate their child, with greater hesitancy for younger children and among younger or less educated parents. Children of vaccinated parents and of parents who thought most of their child’s friends were vaccinated were 4.7 and 1.9 times, respectively, more likely to be vaccinated; unvaccinated parents were 3.2 times more likely to accept the vaccine for their child if they believed most of their friends would vaccinate their children. Further, parents who reported that most of their friends were vaccinated were 1.9 times more likely to have obtained the vaccine themselves, illustrating the influence of social norms. Regardless of their own vaccination status, parents of unvaccinated children were more likely to be politically conservative. If communities or circles of friends could achieve or convey a vaccinated norm, this might persuade undecided or reluctant parents to vaccinate their children. Future research should examine the effects of community behavior and messages highlighting social norms on pediatric vaccine uptake.

New cloning vectors have been developed with features to enhance quick allelic exchange in gram-negative bacteria. The conditionally replicative R6K and transfer origins facilitate conjugation and chromosomal integration into a variety of bacterial species, whereas the sacB gene provides counterselection for allelic exchange. The vectors have incorporated the lacZ alpha fragment with an enhanced multicloning site for easy blue/white screening and priming sites identified for efficient in vivo assembly or other DNA assembly cloning techniques. Different antibiotic resistance markers allow versatility for use with different bacteria, and transformation into an Escherichia coli strain capable of conjugation enables a quick method for allelic exchange. As a proof of principle, the authors used these vectors to inactivate genes in Vibrio cholerae and Salmonella typhimurium.

Both agonism and antagonism of the glucose-dependent insulinotropic polypeptide receptor (GIPR) lead to weight loss in combination with glucagon-like peptide-1 receptor agonists in preclinical models. Here the authors show that this may be explained by desensitization of GIPR activity by chronic GIPR agonism in vitro and in vivo.

The offshore wind energy sector has rapidly expanded over the past two decades, providing a renewable energy solution for coastal nations. Sector development has been led in Europe, but is growing globally. Most developments to date have been in well-mixed, i.e., unstratified, shallow-waters near to shore. Sector growth is, for the first time, pushing developments to deep water, into a brand new environment: seasonally stratified shelf seas. Seasonally stratified shelf seas, where water density varies with depth, have a disproportionately key role in primary production, marine ecosystem and biogeochemical cycling. Infrastructure will directly mix stratified shelf seas. The magnitude of this mixing, additional to natural background processes, has yet to be fully quantified. If large enough it may erode shelf sea stratification. Therefore, offshore wind growth may destabilize and fundamentally change shelf sea systems. However, enhanced mixing may also positively impact some marine ecosystems. This paper sets the scene for sector development into this new environment, reviews the potential physical and environmental benefits and impacts of large scale industrialization of seasonally stratified shelf seas and identifies areas where research is required to best utilize, manage, and mitigate environmental change.

Across Southeast Asia and China, more than 17000 Asian bears are kept under suboptimal conditions and farmed for their bile to meet the consumer demand for traditional medicine products. Years of unsterile and repetitive bile extraction contribute to the development of chronic sterile or bacterial cholecystitis, a pathology commonly diagnosed in formerly bile-farmed bears. In both human and veterinary medicine, the diagnostic value of the macroscopic bile examination for assessing gallbladder disease is unclear. The objective of this study is to identify the role of gallbladder bile color, viscosity, and turbidity, while comparing them with established markers of cholecystitis. Moreover, it aims to define the optimal duration of oral antibiotic treatment for chronic bacterial cholecystitis in bears associated with bile farming. Thirty-nine adult, formerly bile-farmed Asiatic black bears (Ursus thibetanus) were examined under anesthesia and underwent percutaneous ultrasound guided cholecystocentesis. A total of 59 bile samples were collected with 20 animals sampled twice to evaluate the therapeutic success. All bile aspirates were assessed macroscopically and microscopically followed by submission for bacterial culture and antimicrobial sensitivity. In the majority of bears, samples with cytological evidence of bactibilia lacked inflammatory cells and did not always correlate with positive bacterial cultures. The most common bacterial isolates were Enterococcus spp, Streptococcus spp and Escherichia coli. Based on our findings, the optimal duration of antibiotic treatment for chronic bacterial cholecystitis is 30 days. Moreover, unlike Gamma-glutamyl Transferase (GGT) and gallbladder wall thickness, the organoleptic properties of bile were found to be reliable markers of chronic gallbladder inflammation with color and turbidity indicating cholestasis. The current study highlights the importance of cholecystocentesis for the management of gallbladder disease and provides initial results on the possible diagnostic value of macroscopic bile examination.