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Abstract Parkinson’s disease (PD) is a common neurodegenerative disorder with a significant risk proportion driven by genetics. While much progress has been made, most of the heritability remains unknown. This is in-part because previous genetic studies have focused on the contribution of single nucleotide variants. More complex forms of variation, such as structural variants and tandem repeats, are already associated with several synucleinopathies. However, because more sophisticated sequencing methods are usually required to detect these regions, little is understood regarding their contribution to PD. One example is a polymorphic CT-rich region in intron 4 of the SNCA gene. This haplotype has been suggested to be associated with risk of Lewy Body (LB) pathology in Alzheimer’s Disease and SNCA gene expression, but is yet to be investigated in PD. Here, we attempt to resolve this CT-rich haplotype and investigate its role in PD. We performed targeted PacBio HiFi sequencing of the region in 1375 PD cases and 959 controls. We replicate the previously reported associations and a novel association between two PD risk SNVs (rs356182 and rs5019538) and haplotype 4, the largest haplotype. Through quantitative trait locus analyzes we identify a significant haplotype 4 association with alternative CAGE transcriptional start site usage, not leading to significant differential SNCA gene expression in post-mortem frontal cortex brain tissue. Therefore, disease association in this locus might not be biologically driven by this CT-rich repeat region. Our data demonstrates the complexity of this SNCA region and highlights that further follow up functional studies are warranted.

Abstract The diversity of chemical and structural attributes of proteins makes it inherently difficult to produce a wide range of proteins in a single recombinant protein production system. The nature of the target proteins themselves, along with cost, ease of use, and speed, are typically cited as major factors to consider in production. Despite a wide variety of alternative expression systems, most recombinant proteins for research and therapeutics are produced in a limited number of systems: Escherichia coli, yeast, insect cells, and the mammalian cell lines HEK293 and CHO. Recent interest in Vibrio natriegens as a new bacterial recombinant protein expression host is due in part to its short doubling time of ≤ 10 min but also stems from the promise of compatibility with techniques and genetic systems developed for E. coli. We successfully incorporated V. natriegens as an additional bacterial expression system for recombinant protein production and report improvements to published protocols as well as new protocols that expand the versatility of the system. While not all proteins benefit from production in V. natriegens, we successfully produced several proteins that were difficult or impossible to produce in E. coli. We also show that in some cases, the increased yield is due to higher levels of properly folded protein. Additionally, we were able to adapt our enhanced isotope incorporation methods for use with V. natriegens. Taken together, these observations and improvements allowed production of proteins for structural biology, biochemistry, assay development, and structure-based drug design in V. natriegens that were impossible and/or unaffordable to produce in E. coli.

IntroductionNadofaragene firadenovec (Ad-IFNα/Syn3) is now approved for BCG-unresponsive bladder cancer (BLCA). IFNα is a pleiotropic cytokine that causes direct tumor cell killing via TRAIL-mediated apoptosis, angiogenesis inhibition, and activation of the innate and adaptive immune system. We established an immunocompetent murine BLCA model to study the effects of murine adenoviral IFNα (muAd-Ifnα) gene therapy on cancer cells and the tumor microenvironment using a novel murine equivalent of Nadofaragene firadenovec (muAd-Ifnα).MethodsTumors were induced by instilling MB49 cells into the bladders of mice; luciferase imaging confirmed tumor development. Mice were treated with adenovirus control (Ad-Ctrl; empty vector), or muAd-Ifnα (3x1011 VP/mL), and survival analysis was performed. For single-cell sequencing (scRNAseq) analysis (72h), bladders were harvested and treated with collagenase/hyaluronidase and TrypLE for cell dissociation. Single cells were suspended in PBS/1% FBS buffer; viability was assessed with Vicell cell counter. scRNAseq analysis was performed using 10X genomics 3’ sequencing. Raw RNAseq data were pre-processed using Cell Ranger single-cell software. Seurat (R package) was used to normalize and cluster the scRNA data. Pooled differential gene expression analysis in specific cell clusters was performed with DESeq2.ResultsWe identified 16 cell clusters based on marker expression which were grouped into epithelial (tumor), uroplakin-enriched, endothelial, T-cells, neutrophils, and macrophage clusters. Top differentially expressed genes between muAd-Ifnα and Ad-Ctrl were identified. Within the specific cell clusters, IPA analysis revealed significant differences between muAd-Ifnα and control. IFNα signaling and hypercytokinemia/chemokinemia were upregulated in all clusters. Cell death pathways were upregulated in tumor and endothelial clusters. T-cells demonstrated upregulation of the immunogenic cell death signaling pathway and a decrease in the Th2 pathway genes. Macrophages showed upregulation of PD1/PD-L1 pathways along with downregulation of macrophage activation pathways (alternate and classical). Multiplex immunofluorescence confirmed increased infiltration with macrophages in muAd-Ifnα treated tumors compared to controls. PD1/PD-L1 expression was reduced at 72h.DiscussionThis single-cell analysis builds upon our understanding of the impact of Ad-IFNα on tumor cells and other compartments of the microenvironment. These data will help identify mechanisms to improve patient selection and therapeutic efficacy of Nadofaragene firadenovec.

Abstract The skin is complex with multiple layers serving protective, regulatory, and detective functions. The skin hosts chemicals originating from consumption, synthesis, and the environment. Skin chemicals can provide insight into one's daily routine or their level of safety in a work environment. The goal of this study was to investigate the utility of noninvasive skin swabs to detect drugs in a pharmacy setting and to determine whether drugs are transferred to the skin of pharmacy staff. To answer this question, skin swabs were collected from healthy pharmacy staff workers and healthy non‐pharmacy individuals and analyzed via untargeted liquid chromatography–tandem mass spectrometry (LC–MS/MS). Drugs were annotated through library matching against the GNPS community spectral library. We then used questionnaire data to exclude medications that participants took orally or applied topically and focused on the drugs participants were exposed to in the work setting. Overall, pharmacy staff had a higher number and variety of medications on their skin as compared with healthy individuals who did not work in a pharmacy. In addition, we identified some chemicals such as N,N‐Diethyl‐metatoluamide on a large number of subjects in both experimental and control groups, indicating environmental exposure to this compound may be ubiquitous and long‐lasting.

AbstractIncreasing atmospheric greenhouse gas (GHG) concentrations causes global temperature rising as well as extreme hydrological events. Understanding the effects of climate change-induced severe hydrological events such as flooding and drought on soil properties and subsequent GHG emissions from soil has enabled scientists to optimize different land management and global warming potential mitigation of a system. Flooding and drought dynamics affect soil physicochemical properties and GHG emissions specifically from land use systems that are adjacent to streams like riparian buffers. Drivers of soil GHG emissions and the effect of extreme hydrological events on the GHG flux from riparian systems in three climate zones (Mediterranean, temperate, and tropical/subtropical) were investigated in this review utilizing Web of Science and Scopus databases. Wet soil conditions contribute more to global warming (7.2 Mg CO2eq-C ha−1 h−1) than dry soils (2.1 Mg CO2eq-C ha−1 h−1) in riparian systems. Temperate riparian systems showed the greatest soil N2O emissions (0.7 mg N2O-N m−2 h−1) after flooding events/during wet seasons. The greatest CH4 (12 mg CH4-C m−2 h−1) and CO2 (15.2 g CO2-C m−2 d−1) flux from wet soils were observed in tropical/subtropical riparian systems. Greater soil inorganic N content (NO3- and NH4+) in temperate riparian systems is responsible for the higher soil N2O flux during wet seasons. Intense precipitation events and greater soil carbon content in tropical/subtropical riparian systems contributed to more CO2 and CH4 emissions relative to the temperate and Mediterranean riparian systems.

Meat colour is one of the primary factors affecting the acceptability of retail meat cuts. Therefore, displaying the natural bright-red colour of meat is a major goal of the meat industry. Consumers frequently reject to retail meat cuts with discolouration even at low levels of surface browning and/or darkening, which in turn results in significant food waste and economic loss. Thus, understanding the factors that influence meat colour deviations such as discolouration or darkening is crucial in developing effective strategies to minimise the display colour-related quality issues. Numerous factors influence series of biochemical reactions of post-mortem muscles and the chemical state of myoglobin, which ultimately affects meat colour and colour stability. Recent studies have demonstrated the incidence of dark-cutting beef post-mortem can be attributed to the combination of lower glycolytic potential, less glycolytic metabolites, enzymes and dysregulated energy metabolism. This review will focus on the role of post-mortem bioenergetics and mitochondria dynamics that affect beef colour chemistry.

Background: Point prevalence surveys (PPS) organized by the European Centre for Disease Prevention and Control (ECDC) play a crucial role in assessing healthcare-associated infections (HAIs) and antimicrobial use (AU) in European acute care hospitals. In 2017, a crude prevalence of 28.1% (95% CI 27.3-29.0%) of inpatients receiving at least one antimicrobial was recorded in Belgium (patients ≥65 years: 29.6% (95% CI 28.5-30.7%), < 6 5 years : 26.5% (95%CI 25.3-27.6%)) . Following the challenges posed by the COVID-19 pandemic, the 2022 ECDC-PPS aimed to reassess AU levels. Method: A cross-sectional study was conducted between September and November 2022 in 57 representative acute care Belgian hospital sites (35 mergers), following the ECDC-PPS protocol version 6.0. All patients present in surveyed wards at 8 a.m. on the PPS day and not discharged at that time were included. Infection prevention and control teams collected comprehensive data on hospitals, wards, and AU, including agents and indications. Results: Among the 10,142 included inpatients, 29.3% (95%CI 28.4-30.2) were receiving at least one antimicrobial (patients ≥65 years: 31.1% (95% CI 29.7-32.4%), < 6 5 years : 27.1% (95%CI 25.6-28.6%)). Intensive care units (56.3%), surgical (38.7%), and medical wards (33.1%) demonstrated the highest AU prevalence, while psychiatric wards exhibited the lowest (3.0%). A total of 3,549 antimicrobials were recorded, commonly prescribed for treating community-acquired infections (48.6%) and HAIs (30.3%, including 4.2% of long-term care facility acquired infections), as well as for surgical and medical prophylaxis (12.4 and 6.6%, respectively). Notably, only 22.7% of surgical prophylaxis courses (n=100/440) lasted more than one day. The top three most used antimicrobial agents consisted of amoxicillin in combination with a beta-lactamase inhibitor (J01CR02, 20.0%), cefazolin (J01DB04, 9.8%) and piperacillin in combination with a beta-lactamase inhibitor (J01CR05, 9.6%). The most frequently reported diagnoses for medical antimicrobial treatment were pneumonia (25.7%) and urinary tract infections (17.1%). The reason for AU was available in 80.0% of the medical notes. Conclusion: The 2022 PPS reveals an increased AU prevalence (+1.2%) in Belgian acute care hospitals, especially in patients over 65 years of age (+1.5%). This increase was less pronounced in younger patients (< 6 5y) (+0.6%). Future investigations are crucial to delve into prescription attitudes and modifiable practices, emphasizing the urgent need for robust antimicrobial stewardship programs in these healthcare settings.

Abstract Background Frailty is a state of increased vulnerability from physical, social, and cognitive factors resulting in greater risk of negative health-related outcomes and increased healthcare expenditure. A 36-factor electronic frailty index (eFI) developed in the United Kingdom calculates frailty scores using electronic medical record data. There is currently no standardization of frailty screening in Canadian primary care. In order to implement the eFI in a Canadian context, adaptation of the tool is necessary because frailty is represented by different clinical terminologies in the UK and Canada. In considering the promise of implementing an eFI in British Columbia, Canada, we first looked at the content validation of the 36-factor eFI. Our research question was: Does the eFI represent frailty from the perspectives of primary care clinicians and older adults in British Columbia? Methods A modified Delphi using three rounds of questionnaires with a panel of 23 experts (five family physicians, five nurse practitioners, five nurses, four allied health professionals, four older adults) reviewed and provided feedback on the 36-factor eFI. These professional groups were chosen because they closely work as interprofessional teams within primary care settings with older adults. Older adults provide real life context and experiences. Questionnaires involved rating the importance of each frailty factor on a 0–10 scale and providing rationale for ratings. Panelists were also given the opportunity to suggest additional factors that ought to be included in the screening tool. Suggested factors were similarly rated in two Delphi rounds. Results Thirty-three of the 36 eFI factors achieved consensus (> 80% of panelists provided a rating of ≥ 8). Factors that did not achieve consensus were hypertension, thyroid disorder and peptic ulcer. These factors were perceived as easily treatable or manageable and/or not considered reflective of frailty on their own. Additional factors suggested by panelists that achieved consensus included: cancer, challenges to healthcare access, chronic pain, communication challenges, fecal incontinence, food insecurity, liver failure/cirrhosis, mental health challenges, medication noncompliance, poverty/financial difficulties, race/ethnic disparity, sedentary/low activity levels, and substance use/misuse. There was a 100% retention rate in each of the three Delphi rounds. Conclusions and next steps Three key findings emerged from this study: the conceptualization of frailty varied across participants, identification of frailty in community/primary care remains challenging, and social determinants of health affect clinicians’ assessments and perceptions of frailty status. This study will inform the next phase of a broader mixed-method sequential study to build a frailty screening tool that could ultimately become a standard of practice for frailty screening in Canadian primary care. Early detection of frailty can help tailor decision making, frame discussions about goals of care, prevent advancement on the frailty trajectory, and ultimately decrease health expenditures, leading to improved patient and system level outcomes.

Abstract Despite the increasing availability of tandem mass spectrometry (MS/MS) community spectral libraries for untargeted metabolomics over the past decade, the majority of acquired MS/MS spectra remain uninterpreted. To further aid in interpreting unannotated spectra, we created a nearest neighbor suspect spectral library, consisting of 87,916 annotated MS/MS spectra derived from hundreds of millions of MS/MS spectra originating from published untargeted metabolomics experiments. Entries in this library, or “suspects,” were derived from unannotated spectra that could be linked in a molecular network to an annotated spectrum. Annotations were propagated to unknowns based on structural relationships to reference molecules using MS/MS-based spectrum alignment. We demonstrate the broad relevance of the nearest neighbor suspect spectral library through representative examples of propagation-based annotation of acylcarnitines, bacterial and plant natural products, and drug metabolism. Our results also highlight how the library can help to better understand an Alzheimer’s brain phenotype. The nearest neighbor suspect spectral library is openly available for download or for data analysis through the GNPS platform to help investigators hypothesize candidate structures for unknown MS/MS spectra in untargeted metabolomics data.

Nanoparticles are promising candidates as direct therapeutics and delivery systems for osteoarthritis treatments, primarily via intraarticular injection, but little is known about the impact on sliding behavior for a soft material surface like cartilage that would be encountered in a joint. Nanoparticle additives have primarily been studied in the context of hard material interfaces, such as metals or metal oxides, where different lubricating or anti-wear mechanisms depend sensitively on chemical composition, size, and concentration. To understand what nanoparticle parameters influence in situ (in a fluid environment) frictional behavior of soft materials, polyacrylamide (PAM) hydrogels were used as a model soft material platform. Friction tests were conducted in a rheometer with a tribology adapter, with PAM hydrogels molded in a petri dish and immersed in different nanoparticle containing fluid environments. A range of nanoparticle compositions were selected to compare broad categories: gold (metal) with a citrate capping ligand, nanodiamond (carbon), and zirconium dioxide (metal oxide). Comparing surface chemistry, concentration, and degree of aggregation, both nanoparticle surface chemistry and nanoparticle solution viscosity were found to modulate in situ hydrogel friction.

Abstract Introduction Primary spontaneous pneumothorax (PSP) is the presence of air in the pleural space, occurring in the absence of trauma and known lung disease. Standardized expert guidelines on PSP are needed due to the variety of diagnostic methods, therapeutic strategies and medical and surgical disciplines involved in its management. Methods Literature review, analysis of the literature according to the GRADE (Grading of Recommendation, Assessment, Development and Evaluation) methodology; proposals for guidelines rated by experts, patients and organizers to reach a consensus. Only expert opinions with strong agreement were selected. Results A large PSP is defined as presence of a visible rim along the entire axillary line between the lung margin and the chest wall and ≥ 2 cm at the hilum level on frontal chest X-ray. The therapeutic strategy depends on the clinical presentation: emergency needle aspiration for tension PSP; in the absence of signs of severity: conservative management (small PSP), needle aspiration or chest tube drainage (large PSP). Outpatient treatment is possible if a dedicated outpatient care system is previously organized. Indications, surgical procedures and perioperative analgesia are detailed. Associated measures, including smoking cessation, are described. Conclusion These guidelines are a step towards PSP treatment and follow-up strategy optimization in France.

This study aimed to develop poly(lactic acid) (PLA) biodegradable film containing sodium nitrite and α-tocopherol, and to examine its effect on dark-cutting beef color attributes. Using a twin-screw extruder, PLA pellets were manufactured as a low-concentration (LC) PLA pellet containing 0.12% sodium nitrite and 0.5% α-tocopherol or a high-concentration (HC) PLA pellet containing 0.6% sodium nitrite and 2.5% α-tocopherol. Extruded PLA pellets were oven-dried before being compressed and molded into film sheets using a hot press. In this study, 7 normal-pH and 7 dark-cutting strip loins were selected from a commercial processor. Loins were sliced into 2.54-cm thick steaks and randomly assigned to the respective loin-type treatments. The packaging treatments include normal-pH polyvinyl chloride (PVC), dark-cutting PVC, dark-cutting vacuum, LC-PLA dark-cutting in vacuum, and HC-PLA dark-cutting in vacuum. Normal-pH steaks were packaged with PVC overwrap, while dark-cutting control steaks were packaged in PVC overwrap and vacuum-sealed. The respective PLA films (LC or HC) were placed directly on the surface of the dark-cutting steak surface before vacuum packaging, resulting in LC-PLA and HC-PLA treatments. All steaks were placed in a simulated retail dis-play maintained at 2±1°C for 6 d, during which their surface color was evaluated every 24 h using a HunterLab spectrophotometer and a trained panel (n=6). Half of the steaks were removed from display on d 5 and cooked to 71°C on a George Foreman grill to evaluate the cooked color. The remaining steaks were evaluated on d 6 for microbial growth and lipid oxidation. A significant increase (P0.05) compared to dark-cutting steaks in control vacuum packages (without nitrite). There were no differences (P>0.05) in aerobic plate count and lipid oxidation between nitrite-embedded and control vacuum dark-cutting treatments. Results indicate that using lower concentrations of sodium nitrite (0.12%) andα-tocopherol (0.5%) in PLA films can help improve the surface color of raw dark-cutting steaks while minimizing unpleasant cooked color associated with nitrite-embedded film.

The parasitic species Perkinsus olseni (= atlanticus) (Perkinsea, Alveolata) infects a wide range of mollusc species and is responsible for mortality events and economic losses in the aquaculture industry and fisheries worldwide. Thus far, most studies conducted in this field have approached the problem from a “one parasite-one disease” perspective, notably with regards to commercially relevant clam species, while the impact of other Perkinsus species should also be considered as it could play a key role in the disease phenotype and dynamics. Co-infection of P. olseni and P. chesapeaki has already been sporadically described in Manila clam populations in Europe. Here, we describe for the first time the parasitic distribution of two Perkinsus species, P. olseni and P. chesapeaki, in individual clam organs and in five different locations across Arcachon Bay (France), using simultaneous in situ detection by quantitative PCR (qPCR) duplex methodology. We show that P. olseni single-infection largely dominated prevalence (46–84%) with high intensities of infection (7.2 to 8.5 log-nb of copies. g−1of wet tissue of Manila clam) depending on location, suggesting that infection is driven by the abiotic characteristics of stations and physiological states of the host. Conversely, single P. chesapeaki infections were observed in only two sampling stations, Ile aux Oiseaux and Gujan, with low prevalences 2 and 14%, respectively. Interestingly, the co-infection by both Perkinsus spp., ranging in prevalence from 12 to 34%, was distributed across four stations of Arcachon Bay, and was detected in one or two organs maximum. Within these co-infected organs, P. olseni largely dominated the global parasitic load. Hence, the co-infection dynamics between P. olseni and P. chesapeaki may rely on a facilitating role of P. olseni in developing a primary infection which in turn may help P. chesapeaki infect R. philippinarum as a reservoir for a preferred host. This ecological study demonstrates that the detection and quantification of both parasitic species, P. olseni and P. chesapeaki, is essential and timely in resolving cryptic infections and their consequences on individual hosts and clam populations.

OBJECTIVES/GOALS: A primary care clinic is opening in the Eva Rothwell Center (ERC) located in the Keith Neighbourhood of Hamilton, Ontario. This new clinic aims to address rampant health disparities in the community. Effective delivery of health services requires a robust model of care that meets and sustains the specific needs of the community and clinic providers. METHODS/STUDY POPULATION: The primary objective of this study is to describe the current health needs of Keith Neighbourhood residents. The secondary objective is to describe the needs of health providers working within the ERC health clinic. Data collection will involve the collection of primary data (through methods such as surveys and interviews) and secondary data (including historical and current census data). Primary data analysis for primary will use conventional quantitative and qualitative methods. Finally, researchers will use the analyzed data to develop a context-appropriate initial model of care for the ERC health clinic and a subsequent evaluation plan to assess the model’s effectiveness and sustainability. RESULTS/ANTICIPATED RESULTS: All primary and secondary data will be synthesized into a report that will inform the development and implementation of two main deliverables. The first deliverable will be a framework for the clinic’s initial model of care that is context-appropriate to the current needs of the Keith Neighbourhood. The model of care will be culturally sensitive and trauma-informed. The second deliverable will be an evaluation plan for the clinic that can be used to continuously iterate on the initial model, ensuring its sustainability. Furthermore, the project’s process may be extrapolated into a framework that could be used to establish primary care clinics within other priority communities. DISCUSSION/SIGNIFICANCE: The proposed model of care will enable practitioners to deliver relevant and timely health services while being adaptable to the community’s evolving needs. It will help improve the Keith Neighbourhood residents' long-term health and social outcomes. This project will contribute to and inform the development of the field of translational science.

Abstract The dominant benthic primary producers in coral reef ecosystems are complex holobionts with diverse microbiomes and metabolomes. In this study, we characterize the tissue metabolomes and microbiomes of corals, macroalgae, and crustose coralline algae via an intensive, replicated synoptic survey of a single coral reef system (Waimea Bay, Oʻahu, Hawaii) and use these results to define associations between microbial taxa and metabolites specific to different hosts. Our results quantify and constrain the degree of host specificity of tissue metabolomes and microbiomes at both phylum and genus level. Both microbiome and metabolomes were distinct between calcifiers (corals and CCA) and erect macroalgae. Moreover, our multi-omics investigations highlight common lipid-based immune response pathways across host organisms. In addition, we observed strong covariation among several specific microbial taxa and metabolite classes, suggesting new metabolic roles of symbiosis to further explore.

When a notable earthquake occurs in the United States, a range of familiar real‐ and near‐real‐time products are produced by the U.S. Geological Survey (USGS) Advanced National Seismic System (ANSS), and made available via the ANSS Comprehensive Earthquake Catalog. For historical and early instrumental earthquakes, similar results and products are developed depending on data availability and event significance, drawing from published later studies. The year 2023 marked the ninetieth anniversary of the 11 March 1933 Long Beach, California, earthquake. This anniversary provided the impetus to update ANSS products, drawing on archived and published data. Here, we describe the updated ShakeMap, shaking recordings and intensities, and retrospective aftershock forecast for the Long Beach, California, earthquake. In effect we have developed standard, modern ANSS products for an earthquake that occurred 90 yr ago. Our results show that the distributions of both the ground motions, anchored by three strong‐motion recordings, and aftershock magnitudes are consistent with expectations for an M 6.4 mainshock in Southern California. We show that, notwithstanding possible limitations, instrumentally recorded accelerations from the closest station are consistent with predicted shaking and directly estimated macroseismic intensities. Updated data products have been added to the USGS event page, where they are available for download. Public‐facing products were also created for the anniversary and are freely available on the USGS website.

Introduction There is growing evidence that sleep is disrupted after stroke, with worse sleep relating to poorer motor outcomes. It is also widely acknowledged that consolidation of motor learning, a critical component of poststroke recovery, is sleep-dependent. However, whether the relationship between disrupted sleep and poor outcomes after stroke is related to direct interference of sleep-dependent motor consolidation processes, is currently unknown. Therefore, the aim of the present study is to understand whether measures of motor consolidation mediate the relationship between sleep and clinical motor outcomes post stroke.Methods and analysis We will conduct a longitudinal observational study of up to 150 participants diagnosed with stroke affecting the upper limb. Participants will be recruited and assessed within 7 days of their stroke and followed up at approximately 1 and 6 months. The primary objective of the study is to determine whether sleep in the subacute phase of recovery explains the variability in upper limb motor outcomes after stroke (over and above predicted recovery potential from the Predict Recovery Potential algorithm) and whether this relationship is dependent on consolidation of motor learning. We will also test whether motor consolidation mediates the relationship between sleep and whole-body clinical motor outcomes, whether motor consolidation is associated with specific electrophysiological sleep signals and sleep alterations during subacute recovery.Ethics and dissemination This trial has received both Health Research Authority, Health and Care Research Wales and National Research Ethics Service approval (IRAS: 304135; REC: 22/LO/0353). The results of this trial will help to enhance our understanding of the role of sleep in recovery of motor function after stroke and will be disseminated via presentations at scientific conferences, peer-reviewed publication, public engagement events, stakeholder organisations and other forms of media where appropriate.Trial registration number ClinicalTrials.gov: NCT05746260, registered on 27 February 2023.

Objectives: This study aimed to develop a valid and reliable food service satisfaction questionnaire for use by family members of residents in nursing homes. Design: Questionnaire development and validation study conducted using COSMIN® benchmarks for excellence. Setting: Nursing homes. Participants: Family members of residents in nursing homes. Measurements: Content validity was established based on a review of the literature, qualitative interviews with family members (n = 9) and expert review (n = 10). Face validity was established in pilot testing with family members (n = 5). A 40-item questionnaire was developed for psychometric testing. Data from 414 family members was used to establish construct validity (Principal Components Analysis), with data from 101 of those family members used to evaluate reliability (internal consistency (Cronbach’s alpha [α]) and temporal stability [weighted Kappa]). Results: A two factor 24-item scale resulted, with excellent internal consistency (factor one: 11 items related to food and meals (α = 0.960); factor two: 13 items related to the mealtime experience (α = 0.907)). Temporal stability was moderate to near perfect (weighted Kappa: 0.461−0.875; p

Previous research indicated that inherently present electron donors and carriers contribute to nonenzymatic metmyoglobin reduction. However, limited knowledge is currently available on how light and oxygen influence nonenzymatic metmyoglobin reduction. The objective of this study was to evaluate the effect of light and oxygen levels on nonenzymatic metmyoglobin reduction in vitro. Two experiments were conducted using equine metmyoglobin solution at pH 5.6 (represents typical postmortem muscle pH) with different electron donors and electron carriers in a 96-well plate. A gas blend of 80% oxygen and 20% carbon dioxide or a pure 99.9% nitrogen gas was bubbled in phosphate buffers and myoglobin solutions to create 2 oxygen levels (low oxygen [4%] and high oxygen [160%]). The desired dissolved oxygen content was measured using a NeoFox oxygen probe (atmospheric oxygen level was 20% oxygen). There were significant effects of electron donors, carriers, and light on nonenzymatic metmyoglobin reduction. Ascorbate and nicotinamide adenine dinucleotide reduced form (NADH) in the presence of methylene blue and ethylenediaminetetraacetic acid (EDTA) exhibited a greater nonenzymatic metmyoglobin reduction in light than dark. An increase in nonenzymatic metmyoglobin-reducing activity (P

Abstract Background West Nile virus (WNV) is the leading cause of mosquito-borne illness in the continental USA. WNV occurrence has high spatiotemporal variation, and current approaches to targeted control of the virus are limited, making forecasting a public health priority. However, little research has been done to compare strengths and weaknesses of WNV disease forecasting approaches on the national scale. We used forecasts submitted to the 2020 WNV Forecasting Challenge, an open challenge organized by the Centers for Disease Control and Prevention, to assess the status of WNV neuroinvasive disease (WNND) prediction and identify avenues for improvement. Methods We performed a multi-model comparative assessment of probabilistic forecasts submitted by 15 teams for annual WNND cases in US counties for 2020 and assessed forecast accuracy, calibration, and discriminatory power. In the evaluation, we included forecasts produced by comparison models of varying complexity as benchmarks of forecast performance. We also used regression analysis to identify modeling approaches and contextual factors that were associated with forecast skill. Results Simple models based on historical WNND cases generally scored better than more complex models and combined higher discriminatory power with better calibration of uncertainty. Forecast skill improved across updated forecast submissions submitted during the 2020 season. Among models using additional data, inclusion of climate or human demographic data was associated with higher skill, while inclusion of mosquito or land use data was associated with lower skill. We also identified population size, extreme minimum winter temperature, and interannual variation in WNND cases as county-level characteristics associated with variation in forecast skill. Conclusions Historical WNND cases were strong predictors of future cases with minimal increase in skill achieved by models that included other factors. Although opportunities might exist to specifically improve predictions for areas with large populations and low or high winter temperatures, areas with high case-count variability are intrinsically more difficult to predict. Also, the prediction of outbreaks, which are outliers relative to typical case numbers, remains difficult. Further improvements to prediction could be obtained with improved calibration of forecast uncertainty and access to real-time data streams (e.g. current weather and preliminary human cases). Graphical Abstract

In 2020, our research collective facilitated a photovoice project titled “Picturing Our Realities: Arts-based Reflections with Central American Youth in Canada,” which brought together young, second-generation, and one-and-a-half-generation (born in another country and moved at a young age) Central American identifying people in Toronto to talk about their experiences growing up as children of immigrants. This photovoice project reveals the ways the civil war and migration process is a haunting presence in the lives of second and 1.5 generation Central American Canadians as they grow up and carve their paths as adults. We can see how unresolved social conflict emerges and shapes family memory, sense of self, understandings of community, and means of engaging in community activism and community work. We argue that this act of remembering and paying homage to previous generations is a means of confronting and resisting past injustices and forming ways of healing from the afterlives of violence. This recognition of the afterlives of mass violence, and the calls of action that this recognition entails, may form a powerful catalyst for community organizing and creating community spaces to respond to historical hauntings and structural violence.

Wastewater injection has induced earthquakes in Northeastern Colorado since 2014. We apply ambient noise correlation techniques to determine temporal changes in seismic velocities in the region. We find no clear correlation between seismic velocity fluctuations and either injection volumes or seismicity patterns. We do observe apparent annual variations in velocity that may be associated with hydrologic loading or thermoelastic strain. In addition, we model uniform and vertically localized velocity perturbations, and measure the velocity change with 1D synthetic seismograms. Our results indicate that our methods underestimate the known velocity change, especially at shorter station distances and when variations are restricted to a horizontal layer. If injection does cause measurable velocity changes, its effect is likely diluted in cross correlations due to its localized spatial extent around injection wells.

Abstract The COVID-19 pandemic has forced many throughout the world to isolate themselves from their respective communities to stop the spread of disease. Although this form of distancing can prevent the contraction of a virus, it results in social isolation and physical inactivity. Consequently, our communities have become heavily reliant on digital solutions to foster social connection and increase physical activity when forced to isolate. Dance is a multidimensional form of physical activity that includes sensory, motor, cognitive, rhythmic, creative, and social elements. Long-term, interventional studies in dance have shown positive effects on both mental and social health; however, little has been done to examine the acute effects and no studies to date have explored the relationship between the affective state and social outcomes of dance. We examined the hypothesis that online dance is associated with improvements in affective state and social connection during a time of social isolation, namely, the COVID-19 crisis. Healthy adults (age ≥ 18; n = 47) engaged in a single session of 60 min of self-selected online dance, completing a series of validated self-reported questionnaires before and after class. We found that online dance was associated with improvements in affective state as measured by increased positive affect and self-esteem and decreased negative affect and depressive symptoms. Additionally, online dance was associated with improvements in social and community connectedness. Further, we found that those who experienced the largest increases in self-esteem and decreases in negative affect demonstrated the largest gains in social connectivity. Although in-person dance classes may be optimal for formalized dance training, online dance instruction offers an accessible platform that can provide mental and social health benefits during the COVID-19 social isolation crisis. We conclude that through online dance, individuals can experience a connection between the body, mind, and community.

Objective Spinal fractures often have devastating sequelae. Thoracolumbar fractures are classified using the Thoracolumbar Injury Classification and Severity score (TLICS) to determine the severity of injury and to guide treatment. Recently advancements in minimally invasive spine surgery (MISS) have led to new approaches to high-severity fractures. Studies have suggested that MISS may yield similar outcomes to conventional, more invasive procedures while producing several benefits. Methods This retrospective study involves 46 patients treated from 2005 through 2020 for high grade thoracolumbar trauma from T2 to L5 with a minimum follow-up of 6-months treated with MISS techniques using percutaneous instrumentation. Results Average TLICS was 7.5. Patient derived outcome measures with average length of follow-up of 602 days included Oswestry Disability Index 28.9, Patient Satisfaction Index 4.2, Short Form-12 Mental Component Score 51.9, and Short Form-12 Physical Component Score 37.7. Average estimated blood loss was 119.2 mL. Conclusion The TLICS is a validated tool used to guide surgical intervention in high grade trauma. The utilization of MISS techniques for the treatment and stabilization of thoracolumbar trauma is efficacious and a viable alternative to traditional open approaches.

Background Cisgender women represent over half of people living with HIV globally. However, current research efforts toward a cure for HIV focus predominantly on cisgender men. The under-representation of women in HIV cure clinical studies is particularly problematic given data suggesting that sex-dependent phenotypes limit scientific discovery. Objective We aimed to generate considerations to increase the meaningful involvement of women in HIV cure-related research. Materials and methods We conducted in-depth interviews with biomedical researchers and community members to better understand factors that could increase the meaningful involvement of women in HIV cure clinical trials. Participants were affiliated with academia, industry, community advisory boards, and community-based organizations, and were identified using listings from the AIDS Clinical Trials Group and the Martin Delaney Collaboratories. We used conventional content analysis to analyze the qualitative data. Results We recruited 27 participants, of whom 11 were biomedical researchers and 16 were community members. Participants included 25 cisgender women, 1 transgender woman, and 1 cisgender man. Key considerations emerged, including the need to ensure that HIV cure studies reflect HIV epidemiologic trends and having accurate representation by sex and gender in HIV cure research. To increase the meaningful involvement of women, recommendations included instituting intentional enrollment goals, frequent and mandatory reporting on enrollment, and incentives for sites to enroll women. Additional themes included the need for agency and self-determination, attention to lived experiences, trauma and healing, and adequate support for women (e.g. logistical, psychosocial, mental, emotional, and physical). Participants noted that women would be willing to participate in HIV cure trials, related procedures (e.g. biopsies), and analytical treatment interruptions. They also expressed a desired for women-centered and holistic clinical trial designs that account for intersectionality. Conclusions Our empirical inquiry extends recent calls to action to increase diversity of people involved in HIV cure research. Redressing the under-inclusion of women in HIV cure research is an urgent imperative. The entire field must mobilize and reform to achieve this goal. Meaningfully involving women across the gender spectrum in HIV cure research is needed to ensure that interventions are safe, effective, scalable, and acceptable for all people with HIV.

Introduction: Generating physiologically relevant red blood cell extracellular vesicles (RBC-EVs) for mechanistic studies is challenging. Herein, we investigated how to generate and isolate high concentrations of RBC-EVs in vitro via shear stress and mechanosensitive piezo1 ion channel stimulation.Methods: RBC-EVs were generated by applying shear stress or the piezo1-agonist yoda1 to RBCs. We then investigated how piezo1 RBC-EV generation parameters (hematocrit, treatment time, treatment dose), isolation methods (membrane-based affinity, ultrafiltration, ultracentrifugation with and without size exclusion chromatography), and storage conditions impacted RBC-EV yield and purity. Lastly, we used pressure myography to determine how RBC-EVs isolated using different methods affected mouse carotid artery vasodilation.Results: Our results showed that treating RBCs at 6% hematocrit with 10 µM yoda1 for 30 min and isolating RBC-EVs via ultracentrifugation minimized hemolysis, maximized yield and purity, and produced the most consistent RBC-EV preparations. Co-isolated contaminants in impure samples, but not piezo1 RBC-EVs, induced mouse carotid artery vasodilation.Conclusion: This work shows that RBC-EVs can be generated through piezo1 stimulation and may be generated in vivo under physiologic flow conditions. Our studies further emphasize the importance of characterizing EV generation and isolation parameters before using EVs for mechanistic analysis since RBC-EV purity can impact functional outcomes.

Extracts from Euglena gracilis have been shown to prevent cancer growth in mouse models. However, the molecular mechanism of this anti-cancer activity has not been determined nor has the effect of Euglena extracts on tobacco smoke carcinogen-induced carcinogenesis. Here, we investigate the hypothesis that this anti-cancer activity is a result of changes in the intestinal microbiota induced by oral administration of the extract. We found that a Euglena gracilis water extract prevents lung tumorigenesis induced by a tobacco smoke-specific carcinogen (NNK) in mice treated either 2 weeks before or 10 weeks after NNK injection. Both of these treatment regimens are associated with significant increases in 27 microbiota metabolites found in the mouse feces, including large increases in triethanolamine, salicylate, desaminotyrosine, N-acetylserine, glycolate, and aspartate. Increases in the short-chain fatty acids (SCFAs) including acetate, propionate and butyrate are also observed. We also detected a significant attenuation of lung carcinoma cell growth through the induction of cell cycle arrest and apoptosis caused by low levels of SCFAs. This study provides strong evidence of anti-cancer activity in Euglena gracilis extracts against tobacco smoke carcinogen-induced tumorigenesis and demonstrates that this activity is linked to increased production of specific gut microbiota metabolites and the resultant induction of cell cycle arrest and apoptosis of lung carcinoma cells.

High meat pH leads to an undercooked or an abnormal pink appearance in fully-cooked product. High-pressure processing (HPP) promotes protein denaturation. The objective was to evaluate the effects of HPP on cooked steak color and sensory attributes of dark-cutting beef. USDA Choice (mean pH = 5.5) and dark-cutting (mean pH = 6.3) strip loin sections were vacuum packaged and treated with 0 (no HPP), 300, 450, and 600 MPa of pressure for 90 s using chilled water. Steaks were randomly assigned to measure external cooked color, Warner-Bratzler shear force, and trained sensory panel. Applying 300 MPa of pressure improved (P 0.05) a* and chroma of the external cooked steak color. HPP treatments also did not affect (P > 0.05) initial juiciness, sustained juiciness, beef flavor intensity, or overall acceptability. However, 600 MPa made dark-cutting steaks tougher and lighter (P

Tinnitus is prevalent among patients suffering from Single-Sided Deafness (SSD) and Asymmetrical Hearing Loss (AHL). In addition to bothersome tinnitus in the poorer ear, these patients also report issues with understanding speech in noise and sound localization. The conventional treatment options offered to these patients to improve auditory abilities are cochlear implantation, bone conduction devices or Contralateral Routing Of Signal (CROS) hearing aids. It was recently found that the benefit of cochlear implantation for tinnitus associated with AHL/SSD was greater than the other two approaches. It is conceivable that the lack of stimulation provided to the poorer ear in these last approaches explains their modest impact on tinnitus perception. A new technology that combines the ability to reroute the sound from the poorer ear to the good ear (CROS system) while still stimulating the poorer ear with conventional sound amplification has recently been developed: the StereoBiCROS system. The aim of this study was to investigate the effects of this new device on tinnitus. Twelve AHL and two SSD patients aged 70.7 ± 7.9 years with tinnitus were fitted with bilateral hearing aids that included 3 programs: Stereophonic, BiCROS and StereoBiCROS (CROS + bilateral amplification). The short-and long-term effect of the approach on tinnitus was assessed using a tinnitus Loudness Visual Analog Scale (VAS) and the Tinnitus Handicap Inventory (THI), respectively. Both the VAS and the THI were used before and one month after the hearing aid fitting. Of the 14 patients who used their hearing aids daily (12.6 ± 1.6 h per day) the StereoBiCROS program was the most used program (81.8 ± 20.5% of the time). The average THI total score decreased from 47 (± 22) to 15 (± 16) (p = 0.002) and the VAS-Loudness score decreased from 7 (± 1) to 2 (± 2) (p

The objective was to evaluate the effects of high-pressure processing (HPP) levels on retail color of dark-cutting beef. Eight USDA Choice (mean pH=5.5; normal-pH beef) and 12 dark-cutting (mean pH=6.3) strip loins were obtained from a commercial packing plant within 2 d of harvest. Dark-cutting loins were cut into equal sections, vacuum packaged,and randomly assigned to 0 (no HPP), 300, 450, and 600 MPa of pressure for 90 s using chilled water. Following 48 h of dark storage at 2°C, dark-cutting loin sections were cut into 1.9-cm-thick steaks, placed in Styrofoam trays overwrapped in polyvinyl chloride (PVC) film, and placed in a simulated retail display for 8 d. The surface color readings were measured every 24 h using a HunterLab MiniScan XE Plus spectrophotometer, whereas a trained color panel (n=6) evaluated dis-coloration, paleness, and lean color on steaks. Lipid oxidation was evaluated on day 0, 4, and 8 of retail display, and structural changes were determined using light microscopy on day 0 of display. There was a significant HPP level×day of retail display interaction for all instrumental color measurements. Throughout the retail display, L* values of 450 and 600 MPa applied steaks were greater (P

Background Adolescents increasingly access mental health services in schools. School mental health professionals (SPs; school counselors, social workers, etc.) can offer evidence-based mental health practices (EBPs) in schools, which may address access gaps and improve clinical outcomes. Although some studies have assessed factors associated with EBP adoption in schools, additional research focusing on SP- and school-level factors is warranted to support EBP implementation as SPs’ mental health delivery grows. Methods Baseline data were collected from SPs at Michigan high schools participating in a statewide trial to implement SP-delivered cognitive behavioral therapy (CBT) to students. Models examined factors associated with attitudes about EBPs, implementation climate, and implementation leadership, and their associations with CBT knowledge, training attendance, and pre-training CBT delivery. Results One hundred ninety-eight SPs at 107 schools (87%) completed a baseline survey. The mean Evidence-Based Practice Attitude Scale (EBPAS) total score was 2.9, and school-aggregated mean scores of the Implementation Climate Scale (ICS) and Implementation Leadership Scale (ILS) were 1.83 and 1.77, respectively, all on a scale ranging from 0 ( low ) to 4 ( high ). ICS and ILS scores were lower than typically reported in clinical settings, while EBPAS scores were higher. School characteristics were not significantly associated with EBPAS, ICS, or ILS scores, but scores did differ by SP role. Higher EBPAS scores were associated with more CBT knowledge (average marginal effect for 1 SD change [AME] = 0.15 points) and a higher probability of training completion (AME = 8 percentage points). Higher ICS scores were associated with a higher probability of pre-training CBT delivery (AME = 6 percentage points), and higher ILS scores were associated with higher probability of training completion (AME = 10 percentage points). Conclusions Our findings suggest that SPs’ attitudes toward EBPs and organizational support were positively associated with early signs of implementation success. As schools increasingly fill the adolescent mental healthcare access gap, efforts to strengthen both provider attitudes toward EBP and strategic organizational factors supporting EBP delivery will be key to encouraging EBP uptake in schools.

Adoption of electronic identification ear tags (EID) and DNA testing by commercial range sheep producers in the Western United States has been low, despite the availability of these technologies for over a decade. Jointly, these technologies offer an approach to provide individual animal performance data to improve flock health, genetic and reproductive management. This project involved a collaboration with five California sheep producers representing a broad geographic range, varying levels of pre-project EID adoption, and diverse operational practices. Tissue samples were collected from, and ear EIDs were placed in, a total of 2,936 rams and their potential lambs. We partnered with a commercial packing company, Superior Farms, to genotype the animals. Superior Farms used a targeted genotyping panel to assign parentage, and link individual animal identification (ID) to camera-graded carcass measurements. This enabled the collection of individual progeny carcass data and provided insight into sire performance, providing for the within-flock identification of prolific sires that were producing lambs with significantly more saleable meat as compared to their flock mates. Overall, almost 91% of lambs were successfully matched to their sire, and prolificacy ranging from 0-135 lambs per ram. There was as much as an $80 difference in the average edible product from camera-graded carcasses derived from lamb groups sired by different rams. A partial budget analysis modeling investment in an EID system coupled with an autodrafter and scale to collect individual weights and improve labor efficiency during processing, and a sheep flip chute to improve worker safety during foot trimmings, yielded a greater than 7:1 return on investment over a five-year time frame. Ideally, the data collection enabled by EIDs and DNA testing would feed into data-driven genetic evaluation programs to enable selection for more productive and profitable animals, and allow the US sheep industry to accelerate the rate of genetic improvement.

Abstract Objective The COVID-19 pandemic has forced many human subjects research to halt in-person activities and pivot to virtual engagement, including Focus Groups (FGs). We highlight learnings from our experience of hosting virtual FGs from our BEhavioral EConomics for Oral health iNnovation (BEECON) study focusing on oral hygiene behaviors among low-income, predominantly Hispanic families, including practical tips and potential pitfalls to avoid for researchers considering virtual engagement. Results There can be particular benefits to holding virtual sessions among minority parents of young children—to provide flexibility, comfort, and reduced logistical barriers for participation—while still facilitating friendly conversation with minimal distractions. However, extensive preparation is needed to ensure smooth execution and minimal distractions.

Required undergraduate diversity courses often expose students to topics and worldviews which may push them out of their comfort zones and prompt dissonance and even resistance. This paper reports on interviews with 68 faculty members across 16 humanities and social science disciplines at five predominantly white institutions in the Southern United States, detailing how they navigated discussion of controversial topics in required diversity courses. Most instructors aimed to expose students to critical social issues yet were concerned that resistance could disturb the learning process. We identified 20 unique strategies for handling controversial topics in class that included proactively establishing community and safety and normalizing conflict, and reactively acknowledging and surfacing multiple perspectives, as well as connecting content to students' lived experiences. Some instructors also reported a lack of controversy or conflict in their classrooms, which they variously attributed to student characteristics or their own disinclination to promote heated discussion - which, we argue, calls into question the breadth and criteria of many institutionally defined diversity course requirements. We conclude the paper with implications for faculty, educational developers, administrators, and institutions.

Acute pyelonephritis (APN) is a relatively common community-acquired infection, particularly in women. The early appropriate antibiotic treatment of this potentially life-threatening infection is associated with improved outcomes. The international management guidelines for complicated urinary tract infections and APN recommend using oral antibiotics with Escherichia coli and other Enterobacterales to fluoroquinolones, trimethoprim-sulfamethoxazole (TMP-SMX), and beta-lactams has left patients without reliable oral antibiotic treatment options for APN. This narrative review proposes using precision medicine concepts to improve empirical antibiotic therapy for APN in ambulatory settings. Whereas resistance rates to a particular antibiotic class may exceed 10% at the population-based level, the predicted antibiotic resistance rates based on patient-specific risk factors fall under 10% in many patients with APN on the individual level. The utilization of clinical tools for the prediction of fluoroquinolones, TMP-SMX, and third-generation cephalosporin resistance improves the ambulatory antibiotic management of APN. It may also reduce the need to switch antibiotic therapy later based on the in vitro antibiotic susceptibility testing results of bacterial isolates in urinary cultures. This approach may mitigate the burden of increasing antibiotic resistance in the community by ensuring that the initial antibiotic prescribed has the highest likelihood of treating APN appropriately.

BackgroundSurvivors of oral cavity and oropharyngeal cancer frequently experience difficulties in swallowing; tasting; speaking; chewing; and maintaining comfortable movements of the head, neck, and shoulder. Engagement in regular self-care can reduce further loss of function and mitigate late effects. Despite the substantial self-care requirements, there are no empirically based interventions to enhance the skills and confidence of these survivors in managing their ongoing care. ObjectiveThe aim of this study is to describe the rationale and methodology for a randomized controlled trial evaluating Empowered Survivor (ES) versus Springboard Beyond Cancer, a general web-based program for cancer survivors, on self-efficacy in managing care, preparedness for managing survivorship, and health-related quality of life (QOL). MethodsThis study will recruit a total of 600 individuals who were diagnosed with oral cavity or oropharyngeal cancer in the past 3 years and are currently cancer free primarily from state cancer registries; these individuals will be randomly assigned to either the ES or Springboard Beyond Cancer condition. The participants complete measures of self-efficacy in managing care, preparedness for survivorship, health-related QOL, and engagement in oral self-examination and head and neck strengthening and flexibility exercises at baseline and 2 and 6 months after baseline. The primary aim of this study is to evaluate the impact of ES versus Springboard Beyond Cancer on self-efficacy, preparedness, and health-related QOL. The secondary aim is to examine the mediators and moderators of ES’s impact on self-efficacy in managing care, preparedness, and health-related QOL at 6 months. The exploratory aim is to conduct a process evaluation of ES to identify potential oncology or community settings for future implementation. ResultsMultilevel modeling will be used to examine whether there are significant differences between the ES and Springboard Beyond Cancer interventions over time. Mediational models will evaluate the indirect effects of ES on outcomes. Quantitative analyses will evaluate the predictors of ES use, and qualitative analyses will evaluate the preferred timing and settings for the implementation of ES. ConclusionsThis randomized controlled trial evaluates a completely web-based intervention, ES, versus a general web-based program for cancer survivors, Springboard Beyond Cancer, on self-efficacy in managing care, preparedness for managing survivorship, and health-related QOL and identifies the putative mediators and moderators of the intervention’s effects. If an effect on the primary outcomes is illustrated, the next step could be an implementation trial to evaluate the intervention’s uptake in and impact on an oncology care setting or nonprofit organizations. Trial RegistrationClincalTrials.gov NCT04713449; https://clinicaltrials.gov/ct2/show/NCT04713449 International Registered Report Identifier (IRRID)DERR1-10.2196/39996

BackgroundIndividuals diagnosed with melanoma before the age of 40 years (young-onset melanoma survivors) and their first-degree relatives (FDRs) are a growing population at risk for developing recurrent melanoma or new melanomas. Regular surveillance using clinical skin examination (CSE) and skin self-examination (SSE) and engagement in preventive behaviors including sun protection are recommended. Given the growing population of survivors and their families who are at increased risk, it is surprising that no behavioral interventions have been developed and evaluated to improve risk-reduction behaviors. ObjectiveWe describe the rationale and methodology for a randomized controlled trial evaluating the efficacy of a Facebook intervention providing information, goal setting, and peer support to increase CSE, SSE, and sun protection for young-onset melanoma survivors and their FDRs. MethodsOverall, 577 survivors and 577 FDRs will be randomly assigned to either the Young Melanoma Family Facebook Group or the Melanoma Family Healthy Lifestyle Facebook Group condition. Participants will complete measures of CSE, SSE, and sun protection, and mediator measures of attitudes and beliefs before and after the intervention. The primary aim is to evaluate the impact of the Young Melanoma Family Facebook intervention versus the Melanoma Family Healthy Lifestyle Facebook intervention on CSE, SSE frequency and comprehensiveness, and sun protection among FDRs. The secondary aims examine the efficacy of the Young Melanoma Family Facebook intervention on survivors’ SSE frequency and comprehensiveness and sun protection behaviors and mechanisms of intervention efficacy for intervention impact on FDR and survivor outcomes. The exploratory aim is to evaluate the efficacy of the 2 interventions on perceived stress, physical activity, and healthy eating. ResultsThis project was funded by the National Institutes of Health (R01CA221854). The project began in May 2018, and recruitment started in January 2019. We anticipate completing enrollment by November 2023. Power calculations recommended a sample size of 577 survivors and 577 FDRs. Multilevel modeling treating family as the upper-level sampling unit and individual as the lower-level sampling unit will be the primary data analytic approach. Fixed effect predictors in these models will include condition, role, sex, all 2- and 3-way interactions, and covariates. ConclusionsThe Young Melanoma Family Facebook intervention aims to improve primary and secondary skin cancer prevention for young-onset melanoma survivors and their family members. The intervention’s delivery via a popular, freely available social media platform increases its impact because of the potential for dissemination in many contexts. If efficacious, this program could be disseminated by dermatologist practices, public health or nonprofit organizations focused on melanoma, and existing melanoma and skin cancer Facebook groups, thereby expanding its reach. This project will produce a content library of posts and a moderation guide for others. Trial RegistrationClinicalTrials.gov NCT03677739; https://clinicaltrials.gov/ct2/show/NCT03677739 International Registered Report Identifier (IRRID)DERR1-10.2196/39640

A growing public health concern, chronic Diesel Exhaust Particle (DEP) exposure is a heavy risk factor for the development of neurodegenerative diseases like Alzheimer’s (AD). Considered the brain’s first line of defense, the Blood–Brain Barrier (BBB) and perivascular microglia work in tandem to protect the brain from circulating neurotoxic molecules like DEP. Importantly, there is a strong association between AD and BBB dysfunction, particularly in the Aβ transporter and multidrug resistant pump, P-glycoprotein (P-gp). However, the response of this efflux transporter is not well understood in the context of environmental exposures, such as to DEP. Moreover, microglia are seldom included in in vitro BBB models, despite their significance in neurovascular health and disease. Therefore, the goal of this study was to evaluate the effect of acute (24 hr.) DEP exposure (2000 μg/ml) on P-gp expression and function, paracellular permeability, and inflammation profiles of the human in vitro BBB model (hCMEC/D3) with and without microglia (hMC3). Our results suggested that DEP exposure can decrease both the expression and function of P-gp in the BBB, and corroborated that DEP exposure impairs BBB integrity (i.e. increased permeability), a response that was significantly worsened by the influence of microglia in co-culture. Interestingly, DEP exposure seemed to produce atypical inflammation profiles and an unexpected general downregulation in inflammatory markers in both the monoculture and co-culture, which differentially expressed IL-1β and GM-CSF. Interestingly, the microglia in co-culture did not appear to influence the response of the BBB, save in the permeability assay, where it worsened the BBB’s response. Overall, our study is important because it is the first (to our knowledge) to investigate the effect of acute DEP exposure on P-gp in the in vitro human BBB, while also investigating the influence of microglia on the BBB’s responses to this environmental chemical.

Objectives To conceptualize new methods for integrating patient-centered measurement into team-based care. Methods A standalone portal was introduced into a rural clinic to support conceptualization of new methods for integration of patient-centered measurement in team-based care. The portal housed mental health-related online resources, three patient-reported measures and a self-action plan. Six providers and four patients used the portal for four months. Our data collection techniques included clinic discussions, one-on-one interviews, workflow diagrams and data generated through the portal. Analysis was supported through coding interview transcripts, looking across multiple sources of research data and research team discussions. Results Our research team conceptualized five team-based patient-centered measurement methods through this study. Patient-centered measurement Team Mapping offfers a technique to provide greater clarity of care-team roles and responsibilities in data collected through patient-centered measurement. Longitudinal Care Alignment can guide the care-team on incorporating patient-centered measurement into ongoing provider–patient interactions. Digital Tool Exploration can be used to evaluate a team's readiness toward digital tool adoption, and the impact of these tools. Team-based quality improvement serves as a framework for engaging teams in patient-centered quality improvement. Shared learning is a method that promotes patientprovider interactions that validate patient's perspectives of their care. Conclusion The portal illuminated new methods for the integration of patient-centered measurement in team-based care. The first three proposed patient-centered measurement methods provides ways to assess how a clinic can incorporate patient-centered measurement methods into team-based care. The latter two methods focus on the aim of patient-generated data in which patient's values and perspectives are represented and quality of patient-centered care can be evaluated. Further testing is needed to assess the utility of these patient-centered measurement methods across different clinical settings and domains.

Introduction The PLAYshop programme is a novel, brief, theory-based, parent-focused physical literacy intervention in early childhood designed to address the major public health issue of childhood physical inactivity. The primary objective of this study is to examine the efficacy of the virtually delivered PLAYshop programme in increasing preschool-aged children’s physical literacy, including fundamental movement skills and motivation and enjoyment.Methods and analysis This study aims to recruit 130 families with preschool-aged children (3–5 years) from Alberta and British Columbia, Canada who will be randomised to an intervention or control group. The PLAYshop programme is informed by the Capability, Opportunity, Motivation, Behavior (COM-B) model and includes four intervention strategies: (1) educational training via a 60 min virtual synchronous workshop, (2) educational resources via handouts, (3) material resources via a goody bag of basic active play equipment and (4) follow-up support via access to a digital app with an online toolkit and four biweekly booster lessons (1-week, 3-week, 5-week and 7-week follow-up). To assess the primary outcome of physical literacy, five fundamental movement skills (overhand throw, underhand throw, horizontal jump, hop, one leg balance) will be measured virtually at baseline and 2-month follow-up using the Test of Gross Motor Development (TGMD) and the Movement Assessment Battery for Children-Second Edition (MABC-2) tools. Additionally, children’s motivation and enjoyment will also be assessed at baseline and 2-month follow-up by: (1) parental-report using items from the Preschool Physical Literacy Assessment (PrePLAy) and (2) self-report using an adapted Five Degrees of Happiness Likert scale for children. The control group will receive the PLAYshop programme after the 2-month follow-up.Ethics and dissemination The protocol was approved by the University of Alberta (00093764) and University of Victoria (16-444) Research Ethics Boards. Findings will be disseminated through peer-reviewed publications, conference presentations, social and traditional media and a circulated infographic.Trial registration number NCT05255250.

Objectives: Demonstrate the benefits of using 3D printed skull models when counseling families regarding disorders of the cranial vault (namely plagiocephaly and craniosynostosis), as traditional imaging review and discussion is often insufficient. Methods: 3D printed skull models of a patient with plagiocephaly were used during clinic appointments to aid in the counseling of parents. Surveys were distributed following the appointment to evaluate the utility of these models during the discussion. Results: Fifty surveys were distributed (with a 98% response rate). 3D models were both empirically and anecdotally helpful for parents in understanding their child's diagnosis. Conclusion: Advances in 3D printing technology and software have made producing models more accessible. Incorporating physical, disorder-specific models into our discussions has led to improvements in our ability to communicate with our patients and their families. Innovation: Disorders of the cranial can be challenging to describe to the parents and guardians of affected children; using 3D printed models is a useful adjunct in patient-centered discussions. The subject response to the use of these emerging technologies in this setting suggests a major role for 3D models in patient education and counseling for cranial vault disorders.