Targeted RNA editing in brainstem alleviates respiratory dysfunction in a mouse model of Rett syndrome.

Rett syndrome is a neurological disease due to loss-of-function mutations in the transcription factor, Methyl CpG binding protein 2 (MECP2). Because overexpression of endogenous MECP2 also causes disease, we have exploited a targeted RNA-editing approach to repair patient mutations where levels of MECP2 protein will never exceed endogenous levels. Here, we have constructed adeno-associated viruses coexpressing a bioengineered wild-type ADAR2 catalytic domain (Editasewt) and either Mecp2-targeting or nontargeting gfp RNA guides. The viruses are introduced systemically into male mice containing a guanosine to adenosine mutation that eliminates MeCP2 protein and causes classic Rett syndrome in humans. We find that in the mutant mice injected with the Mecp2-targeting virus, the brainstem exhibits the highest RNA-editing frequency compared to other brain regions. The efficiency is sufficient to rescue MeCP2 expression and function in the brainstem of mice expressing the Mecp2-targeting virus. Correspondingly, we find that abnormal Rett-like respiratory patterns are alleviated, and survival is prolonged, compared to mice injected with the control gfp guide virus. The levels of RNA editing among most brain regions corresponds to the distribution of guide RNA rather than Editasewt. Our results provide evidence that a targeted RNA-editing approach can alleviate a hallmark symptom in a mouse model of human disease.

How to use the Harmonising Outcome Measures for Eczema Core Outcome Set for atopic dermatitis trials: a users' guide.

BACKGROUND: The Harmonising Outcome Measures for Eczema (HOME) initiative has agreed upon the Core Outcome Set (COS) for use in atopic dermatitis (AD) clinical trials, but additional guidance is needed to maximize its uptake. OBJECTIVES: To provide answers to some of the commonly asked questions about using the HOME COS; to provide data to help with the interpretation of trial results; and to support sample size calculations for future trials. METHODS AND RESULTS: We provide practical guidance on the use of the HOME COS for investigators planning clinical trials in patients with AD. It answers some of the common questions about using the HOME COS, how to access the outcome measurement instruments, what training/resources are needed to use them appropriately and clarifies when the COS is applicable. We also provide exemplar data to inform sample size calculations for eczema trials and encourage standardized data collection and reporting of the COS. CONCLUSIONS: By encouraging adoption of the COS and facilitating consistent reporting of outcome data, it is hoped that the results of eczema trials will be more comprehensive and readily combined in meta-analyses and that patient care will subsequently be improved.

A core domain set for pyoderma gangrenosum trial outcomes: an international eDelphi and consensus study from the UPGRADE initiative.

BACKGROUND: Pyoderma gangrenosum (PG) is a rare ulcerative skin condition with no current standardized outcomes or outcome measures. With a rich investigational therapeutic pipeline, standardization of outcomes and improvement of data quality and interpretability will promote the appropriate and consistent evaluation of potential new therapies. Core outcome sets (COS) are agreed, standardized sets of outcomes that represent the minimum that should be measured and reported in all clinical trials of a specific condition. OBJECTIVES: To identify and reach a consensus on which domains (what to be measured) should be included in the Understanding Pyoderma Gangrenosum: Review and Analysis of Disease Effects (UPGRADE) core domain set for clinical trials in PG. METHODS: Collaborative discussions between patients and PG experts, and a systematic review of the literature identified items and prospective domains. A three-round international eDelphi exercise was performed to prioritize the domains and refine the provisional items (consensus: ≥ 70% of participants rating a domain as 'extremely important' and < 15% of participants voting 'not important'), followed by an international meeting to reach consensus on the core domain set (consensus: < 30% disagreement). Item-generation discussions and consensus meetings were hosted via online videoconferences. The eDelphi exercise and consensus voting were performed using Qualtrics survey software. Participants were adults with PG, healthcare professionals, researchers and industry representatives. RESULTS: Collaborative discussions and systematic reviews yielded 115 items, which were distilled into 15 prospective domains. The eDelphi exercise removed the three lowest-priority domains ('laboratory tests', 'treatment costs' and 'disease impact on family') and ranked 'pain', 'quality of life' and 'physical symptoms' as the highest-priority prospective domains. Consensus was reached on the domains of 'pain', 'quality of life' and 'clinical signs'. The domain of 'disease course/disease progression' narrowly failed to reach consensus for inclusion in the core set (32% of participants voted 'no'). Refinement of this domain definition will be required and presented for consideration at future consensus meetings. CONCLUSIONS: The UPGRADE core domain set for clinical trials in PG has been agreed by international multistakeholder consensus. Future work will develop and/or select outcome measurement instruments for these domains to establish a COS.

A Randomized, Double-Blind, Placebo-Controlled, Multicenter, 16-Week Trial to Evaluate the Efficacy and Safety of FB-401 in Children, Adolescents, and Adult Subjects (Ages 2 Years and Older) with Mild-to-Moderate Atopic Dermatitis.

BACKGROUND: Atopic dermatitis is a common chronic, relapsing, and remitting inflammatory skin disorder associated with cutaneous dysbiosis. Current treatment options often fail to adequately control the disease and have unfavorable safety profiles. There is a need for new options that address these treatment shortcomings. OBJECTIVE: The aim of the study was to evaluate the efficacy, safety, and tolerability of FB-401, a live therapeutic product of 3 strains of Roseomonas mucosa, compared to matching placebo applied topically 3 times per week to participants ages ≥2 years of age with mild-to-moderate atopic dermatitis. METHODS: A randomized, double-blind, placebo-controlled, parallel-group study was conducted. The primary outcome was the proportion of participants with 50% improvement in Eczema Area and Severity Index score from baseline at week 16. 154 subjects aged 2 or older with a clinical diagnosis of atopic dermatitis as defined by Hanifin and Rajka criteria with mild or moderate severity were randomized 1:1 via interactive web response system to FB-401 or placebo. RESULTS: The proportion of subjects who achieved the primary outcome was similar between both treatment groups, with no significant treatment group differences observed at any post-baseline visit. The number of treatment-emergent adverse events and the number of subjects with at least one were similar across treatment groups. One serious adverse event not related to treatment was reported. No treatment-emergent adverse events led to treatment discontinuation or study discontinuation. CONCLUSIONS: FB-401 showed an acceptable safety profile but failed to prove superior to placebo in treating children and adults with mild-to-moderate atopic dermatitis.

The Harmonising Outcome Measures for Eczema (HOME) implementation roadmap.

BACKGROUND: Core outcome sets (COS) are consensus-driven sets of minimum outcomes that should be measured and reported in all clinical trials. COS aim to reduce heterogeneity in outcome measurement and reporting, and selective outcome reporting. Implementing COS into clinical trials is challenging. Guidance to improve COS uptake in dermatology is lacking. OBJECTIVES: To develop a structured practical guide to COS implementation. METHODS: Members of the Harmonising Outcome Measurement for Eczema (HOME) executive committee developed an expert opinion-based roadmap founded on a combination of a review of the COS implementation literature, the Core Outcome Measures in Effectiveness Trials (COMET) initiative resources, input from HOME members and experience in COS development and clinical trials. RESULTS: The data review and input from HOME members was synthesized into themes, which guided roadmap development: (a) barriers and facilitators to COS uptake based on stakeholder awareness/engagement and COS features; and (b) key implementation science principles (assessment-driven, data-centred, priority-based and context-sensitive). The HOME implementation roadmap follows three stages. Firstly, the COS uptake scope and goals need to be defined. Secondly, during COS development, preparation for future implementation is supported by establishing the COS as a credible evidence-informed consensus by applying robust COS development methodology, engaging multiple stakeholders, fostering sustained and global engagement, emphasizing COS ease of use and universal applicability, and providing recommendations on COS use. Thirdly, incorporating completed COS into primary (trials) and secondary (reviews) research is an iterative process starting with mapping COS uptake and stakeholders' attitudes, followed by designing and carrying out targeted implementation projects. Main themes for implementation projects identified at HOME are stakeholder awareness/engagement; universal applicability for different populations; and improving ease-of-use by reducing administrative and study burden. Formal implementation frameworks can be used to identify implementation barriers/facilitators and to design implementation strategies. The effect of these strategies on uptake should be evaluated and implementation plans adjusted accordingly. CONCLUSIONS: COS can improve the quality and applicability of research and, so, clinical practice but can only succeed if used and reported consistently. The HOME implementation roadmap is an extension of the original HOME roadmap for COS development and provides a pragmatic framework to develop COS implementation strategies.

2022 Optical Interference Coatings Conference: Manufacturing Problem Contest [Invited].

Participants in the 2022 Manufacturing Problem Contest were challenged to fabricate an optical filter with a specified stepped transmittance spanning three orders of magnitude from 400 to 1100 nm. The problem required that contestants be versed in the design, deposition, and measurement of optical filters to achieve good results. Nine samples from five institutions were submitted with total thicknesses between 5.9 and 53.5 µm with between 68 and 1743 layers. The filter spectra were measured by three independent laboratories. The results were presented in June 2022 at the Optical Interference Coatings Conference in Whistler, B.C., Canada.

Pathways and sources of lead exposure: Michigan Children's Lead Determination (the MI CHILD study).

There has been little research on childhood lead exposure pathways since the 1990s. New data from Michigan in 2017-2021 for 429 children in 345 homes included lead in blood, paint, dust, soil, water, and other housing, demographic, and behavioral metrics. Fifty-three percent of these children had blood lead (BPb) ≥5 µg/dL. A repeated measures pathway model that accounted for multiple children in the same home was constructed using weighted least squares mean estimation and included variance-covariance model multiple imputation. Results showed that children's BPb was directly predicted by lead in settled floor house dust, child's age, season, and mouthing behavior and indirectly predicted by window sill and trough dust lead (DPb), bare soil lead (SPb), proportion of floors with carpets, and exterior building deteriorations. Paint lead (PPb) was also an indirect predictor of BPb through the soil and settled dust pathways. Water lead (WPb), water consumption and other lead sources/pathways were not significant predictors of BPb in this cohort. Although risk factors for individual children are highly variable and worthy of investigation to pinpoint their exposures, this study shows that the main direct and indirect pathways of lead exposure for most children in older housing remain paint and the contaminated dust and soil it generates. Pathway analyses in other jurisdictions using current data should be performed to confirm these results. This study suggests both DPb and BPb in high-risk homes may have declined since the 1990s and that lead in dust, soil, and paint all should be measured to predict risk and target remediation. Because most homes still have not been assessed for lead hazards and remediated, too many children remain at needless risk.

The Demise of Artificial Trans Fat: A History of a Public Health Achievement.

Policy Points This article describes a strategic combination of research, advocacy, corporate campaigns, communications, grassroots mobilization, legislation, regulatory actions, and litigation against companies and government to secure a national policy to remove artificial trans fat from the US food system. Sharing lessons we learned can help inform policymakers, academics, policy practitioners, and students across disciplines. Some of our lessons are that system change means that all consumers benefit without the need for individual behavior change; research can both identify opportunities to improve health and support policy adoption; policy efforts can serve as public education campaigns; policy campaigns can drive marketplace changes; and engaging forward-thinking companies can diffuse opposition to passing a policy. CONTEXT: For many decades, partially hydrogenated vegetable oil (PHO), the primary source of artificial trans fat in the American diet, was used widely in processed and restaurant foods. In the early 1990s, studies linked the consumption of artificial trans fat with heart disease. This article details how research and advocacy led to eliminating artificial trans fat from the US food supply. METHODS: We synthesized published studies of the health impact of trans fat, the legislative history of state and local trans fat bills, the Food and Drug Administration's (FDA) regulatory docket on trans fat labeling and its declaration that PHOs are no longer Generally Recognized as Safe (GRAS), and our own files, which included strategy documents, notes from meetings with the FDA staff, correspondence between advocates and the FDA, fact sheets, press releases, news clips, and other materials. FINDINGS: This history of trans fat provides insights into policy strategy and advocacy best practices that resulted in the removal of trans fat from food in the United States, preventing an estimated 50,000 premature deaths a year. The lessons we learned are that system change benefits all consumers without the need for individual behavior change; research can both identify opportunities to improve health through policy and support policy adoption; policy campaigns can serve as public education campaigns; policy can drive changes to products and the marketplace; and engaging forward-thinking companies can help diffuse opposition to passing a policy. Securing this policy required the persistence of scientists and health advocates in first discovering the risks and then using the science to secure policies to mitigate the identified harm. CONCLUSIONS: An understanding of the tactics used to help attain the targeted policies and how challenges were addressed (such as through communications, leveraging an expanding research base and expert reports, showing that a national policy was feasible through voluntary corporate changes and state and local policy, and litigation against companies and government agencies) may provide a model for scientists, students, advocates, and policymakers. We hope this account will inform efforts to address other public health challenges, such as the current threats of excessive exposure to sodium and added sugars, which persist in the US food system.

Policies to solve the salt problem.

Diets high in sodium have long been known to raise blood pressure, which, in turn, increases the risk of cardiovascular disease. Though authoritative recommendations have been made in the past several decades for federal policies and programs to reduce sodium consumption, measures adopted to date have not been effective. We recommend a comprehensive public health approach to reduce sodium in the food supply and prevent thousands of unnecessary deaths and billions of dollars in health-care costs each year.

Effects of obesity and diabesity on heart rhythm in the Zucker rat.

Obesity and type 2 diabetes mellitus are risk factors for hypertension, coronary heart disease, cardiac arrhythmias including atrial fibrillation, heart failure and sudden cardiac death. The effects of obesity and diabesity on heart rhythm were investigated in the Zucker diabetic fatty (ZDF) and Zucker fatty (ZF) compared to the Zucker lean (ZL) control rat. In vivo biotelemetry techniques were used to assess the electrocardiogram and other cardiac and metabolic parameters. ZDF rats were characterized by age-dependent elevations in fasting and non-fasting blood glucose, glucose intolerance and weight gain and ZF rats were characterized by smaller elevations in fasting and non-fasting blood glucose and greater weight gain compared to ZL rats. Heart rate (HR) was progressively reduced in ZDF, ZF and ZL rats. At 195 days (6.5 months) of age there were significant differences in HR between ZDF (265 ± 8 bpm, n = 10), ZF (336 ± 9 bpm, n = 10) and ZL (336 ± 10 bpm, n = 10) rats and significant differences in HRV between ZDF (22 ± 1 bpm, n = 10), ZF (27 ± 1 bpm, n = 10) and ZL (31 ± 1 bpm, n = 10) rats. Power spectral analysis revealed no significant (P > 0.05) differences in HRV at low frequencies, reduced HRV at high frequencies and increased sympathovagal balance in ZDF compared to ZF and ZL rats. HR was reduced by ageing and additionally reduced by diabesity in the absence of changes in physical activity and body temperature. Reductions in HRV associated with altered sympathovagal drive might partly underlie disturbed HR in the ZDF rat. Possible explanations for reduced HR and future mechanistic studies are discussed.

2019 Topical Meeting on Optical Interference Coatings: Manufacturing Problem Contest [invited].

For the seventh Manufacturing Problem Contest, participants were challenged to fabricate an optical filter with transmittance specified for s-polarization at two incident angles: 10° and 50° from 400 nm to 1100 nm. The problem required that contestants be equally versed in the design, deposition, and measurement of optical filters in order to achieve good results. Eight teams from five different countries participated in the contest using various deposition techniques. The fabricated filters had a total thickness between 8.2 µm and 17.6 µm and a total number of layers from 74 to 255, which were deposited onto one or both sides of the substrate. The performances of the filters were measured by two independent laboratories. The evaluation results were presented at the Topical Meeting on Optical Interference Coatings conference held in Santa Ana Pueblo, New Mexico, in June 2019.

Self-reported adverse reactions associated with mycoprotein (Quorn-brand) containing foods.

BACKGROUND: Mycoprotein, which is produced by a mold and is the basis of Quorn-brand meat substitutes, is a novel cause of allergic and gastrointestinal reactions, but little information has been available on its associated symptomatology. OBJECTIVE: To describe the nature and frequency of adverse reactions to mycoprotein. METHODS: Self-reports of adverse reactions to mycoprotein were collected via a Web-based questionnaire (www.quorncomplaints.org) and then analyzed. RESULTS: Analysis of 1,752 adverse reactions found that Quorn products caused allergic and gastrointestinal symptoms, with some people experiencing both. Allergic reactions, including urticaria and anaphylaxis, occurred within 4 hours of consumption in 312 people. Of those reactions, 45.8%, 1 fatal, began within 1 hour of exposure. Of those 312 individuals, 188 (60.3%) reported repeated reactions after repeated consumption of Quorn, and 2 people experienced 8 reactions (13 people did not say whether they experienced more than 1 reaction). Quorn foods caused gastrointestinal symptoms, including emesis and diarrhea, within 8 hours of consumption in 1,692 people. Of the gastrointestinal symptoms, 66.6% occurred 46 to 180 minutes after consumption of the products. Symptoms ranged from mild nausea to emesis severe enough to warrant medical attention. CONCLUSION: Mycoprotein may be causing numerous and sometimes life-threatening allergic and gastrointestinal reactions. The acceptance in the food supply of this nonessential ingredient deserves reconsideration.

Manufacturing problem contest [invited].

For the Manufacturing Problem contest, participants were asked to fabricate on provided blank substrates a challenging filter with specific reflectance and transmittance targets covering a wavelength range from 400 nm to 1100 nm. The problem was selected such that to achieve a performance close to the targets, a submitted filter had to include at least one thin absorbing layer. Nine teams from six countries participated in the contest using different deposition techniques. The teams' designs had a number of layers varying from 36 to 235, and a total thickness from 2.0 µm to 14.6 µm. The performances of all submitted filters were measured by two independent laboratories, and the results were presented at the Optical Interference Coating meeting in June 2016.

Broadband transmission filters from the 2013 Optical Interference Coatings manufacturing problem contest [invited].

A broadband transmission filter from 400 to 1100 nm was selected for the manufacturing problem contest. The purpose of the contest is to test the state of the art of current optical thin film manufacturing capabilities. A total of 37 people from 15 teams participated in the contest and submitted 17 samples. Diverse approaches were taken by participants to tackle the problem. A range of different solutions was obtained where the number of layers varied from 22 to 608, and the total layer thickness ranged from 1.859 to 23.099 µm. Two independent laboratories performed sample evaluation measurements. Three teams shared the best result with the lowest average measured merit function.

Nutritional properties of raw and cooked Azolla caroliniana Willd., an aquatic wild edible plant.

Azolla caroliniana Willd. is an understudied wild edible plant native to the Eastern United States. Other species of Azolla have been used across the world for several thousand years as a livestock feed and as "green manure." The use of Azolla for human consumption is thought to be limited by its high total polyphenolic content (TPC). However, the TPC and nutritional content of A. caroliniana has not been thoroughly studied. We measured TPC and other nutrients before and after cooking methods designed to lower TPC. We found that TPC was 4.26 g gallic acid equivalent (GAE) kg-1 DW in raw A. caroliniana. All cooking methods significantly lowered TPC. Protein content was 19% DW, and the apparent protein digestibility was 78.45%. Our yield was 173 g FW m-2 day-1 and 5.53 g DW m-2 day-1. Azolla caroliniana is a high-yielding plant with great potential for cultivation and domestication.

Measuring Signs of Atopic Dermatitis in Clinical Practice: A HOME-CP Consensus Statement.

Importance: Outcome measurement is an essential component of value-based health care and can aid patient care, quality improvement, and clinical effectiveness evidence generation. The Harmonising Outcome Measures for Eczema Clinical Practice initiative aims to identify a list of validated, feasible, outcome measurement instruments recommended to measure atopic dermatitis (AD) in the clinical practice setting. The clinical practice set is a list of instruments that clinicians can pick and choose from to suit their needs in the context of clinical care. Objective: To recommend instruments to measure clinical signs of AD in clinical practice. Evidence Review: Following the predefined roadmap, a mixed methods design was implemented and incorporated systematic reviews and qualitative consensus methods. Previous systematic reviews identified few clinical signs instruments with sufficient validation for recommendation. An updated systematic review evaluating the validity of clinical signs instruments informed an international meeting to reach consensus on recommended instruments to measure AD clinical signs in clinical practice. Consensus was defined as less than 30% disagreement. An in-person consensus exercise was held in Montreal, Canada, on October 16, 2022. The 34 attendees included patient and patient advocate research partners, health care professionals, researchers, methodologists, and industry representatives. Findings: The updated systematic review found that the Eczema Area and Severity Index (EASI), Scoring Atopic Dermatitis, and objective Scoring Atopic Dermatitis were the only instruments that demonstrated sufficient performance in all assessed measurement properties. The modified EASI and Signs Global Assessment × Body Surface Area instruments were also recommended. The EASI, Validated Investigator Global Assessment, and Investigator's Global Assessment multiplied by or measured concurrently with a body surface area measure achieved consensus in criteria and were adopted. Conclusions and Relevance: This consensus statement by the Harmonising Outcome Measures for Eczema initiative suggests that when assessing and documenting clinical signs of AD, there are several valid and feasible instruments that can best fit a clinician's specific practice needs. These instruments should improve and standardize the documentation of signs severity, help determine the effect of treatment, facilitate the generation of clinical effectiveness evidence, and enhance the implementation of value-based health care.

Trends in trans fatty acids reformulations of US supermarket and brand-name foods from 2007 through 2011.

INTRODUCTION: Although some US food manufacturers have reduced trans fatty acids (TFA) in their products, it is unknown how much TFA is being reduced, whether pace of reformulation has changed over time, or whether reformulations vary by food type or manufacturer. METHODS: In 2007, we identified 360 brand-name products in major US supermarkets that contained 0.5 g TFA or more per serving. In 2008, 2010, and 2011, product labels were re-examined to determine TFA content; ingredients lists were also examined in 2011 for partially hydrogenated vegetable oils (PHVO). We assessed changes in TFA content among the 270 products sold in all years between 2007 and 2011 and conducted sensitivity analyses on the 90 products discontinued after 2007. RESULTS: By 2011, 178 (66%) of the 270 products had reduced TFA content. Most reformulated products (146 of 178, 82%) reduced TFA to less than 0.5 g per serving, although half of these 146 still contained PHVO. Among all 270 products, mean TFA content decreased 49% between 2007 and 2011, from 1.9 to 0.9 g per serving. Yet, mean TFA reduction slowed over time, from 30.3% (2007-2008) to 12.1% (2008-2010) to 3.4% (2010-2011) (P value for trend < .001). This slowing pace was due to both fewer reformulations among TFA-containing products at start of each period and smaller TFA reductions among reformulated products. Reformulations also varied substantially by both food category and manufacturer, with some eliminating or nearly eliminating TFA and others showing no significant changes. Sensitivity analyses were similar to main findings. CONCLUSIONS: Some US products and food manufacturers have made progress in reducing TFA, but substantial variation exists by food type and by parent company, and overall progress has significantly slowed over time. Because TFA consumption is harmful even at low levels, our results emphasize the need for continued efforts toward reformulating or discontinuing foods to eliminate PHVO.

Muscle coordination and recruitment during squat assistance using a robotic ankle-foot exoskeleton.

Squatting is an intensive activity routinely performed in the workplace to lift and lower loads. The effort to perform a squat can decrease using an exoskeleton that considers individual worker's differences and assists them with a customized solution, namely, personalized assistance. Designing such an exoskeleton could be improved by understanding how the user's muscle activity changes when assistance is provided. This study investigated the change in the muscle recruitment and activation pattern when personalized assistance was provided. The personalized assistance was provided by an ankle-foot exoskeleton during squatting and we compared its effect with that of the no-device and unpowered exoskeleton conditions using previously collected data. We identified four main muscle recruitment strategies across ten participants. One of the strategies mainly used quadriceps muscles, and the activation level corresponding to the strategy was reduced under exoskeleton assistance compared to the no-device and unpowered conditions. These quadriceps dominant synergy and rectus femoris activations showed reasonable correlations (r = 0.65, 0.59) to the metabolic cost of squatting. These results indicate that the assistance helped reduce quadriceps activation, and thus, the metabolic cost of squatting. These outcomes suggest that the muscle recruitment and activation patterns could be used to design an exoskeleton and training methods.