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Fractures continue to be a global economic burden as there are currently no osteoanabolic drugs approved to accelerate fracture healing. In this study, we aimed to develop an osteoanabolic therapy which activates the Wnt/β-catenin pathway, a molecular driver of endochondral ossification. We hypothesize that using an mRNA-based therapeutic encoding β-catenin could promote cartilage to bone transformation formation by activating the canonical Wnt signaling pathway in chondrocytes. To optimize a delivery platform built on recent advancements in liposomal technologies, two FDA-approved ionizable phospholipids, DLin-MC3-DMA (MC3) and SM-102, were used to fabricate unique ionizable lipid nanoparticle (LNP) formulations and then tested for transfection efficacy both in vitro and in a murine tibia fracture model. Using firefly luciferase mRNA as a reporter gene to track and quantify transfection, SM-102 LNPs showed enhanced transfection efficacy in vitro and prolonged transfection, minimal fracture interference and no localized inflammatory response in vivo over MC3 LNPs. The generated β-cateninGOF mRNA encapsulated in SM-102 LNPs (SM-102-β-cateninGOF mRNA) showed bioactivity in vitro through upregulation of downstream canonical Wnt genes, axin2 and runx2. When testing SM-102-β-cateninGOF mRNA therapeutic in a murine tibia fracture model, histomorphometric analysis showed increased bone and decreased cartilage composition with the 45 μg concentration at 2 weeks post-fracture. μCT testing confirmed that SM-102-β-cateninGOF mRNA promoted bone formation in vivo, revealing significantly more bone volume over total volume in the 45 μg group. Thus, we generated a novel mRNA-based therapeutic encoding a β-catenin mRNA and optimized an SM-102-based LNP to maximize transfection efficacy with a localized delivery.

Veteran access to sleep medicine is of paramount importance to the Veterans Health Administration (VA). To increase access, VA has created community referral policies and programs, as well as telehealth programs. In 2017, the Office of Rural Health (ORH) funded a TeleSleep initiative focused on reaching rural Veterans with unmet sleep needs. ORH provided 3–6 years of funding to help 19 hubs support 98 spoke sites serving rural Veterans. As ORH funding concluded, each hub identified its path to sustainment. This case study follows one TeleSleep hub in VA's western geographic region as it transitioned from ORH funding sustainment as a regional Sleep Clinical Resource Hub. This case study describes the real-world process of adaptation in care delivery strategies. One key area of adaptation revolved around whether to deliver care via the patient's home facility or the provider's home facility. In early 2021, the TeleSleep team implemented an innovative provider transfer model, where temporary reinforcements from the TeleSleep hub increased the workforce capacity of spoke sites, similar to the concept of locum tenens. In this provider transfer model, TeleSleep clinicians scheduled, documented, and billed for each encounter at the Veteran's home facility. Positioning TeleSleep clinicians as local providers facilitated communication and referrals and promoted continuity and quality of care for Veterans in their home facility. This provider transfer model reduced the administrative burden of providers and schedulers and supported patient-side-only documentation of care. While this mirrors current locum tenens practice, transferring providers did not fit VA's financial model as implemented by the western region's Sleep Clinical Resource Hub. Therefore, in December 2021, VA aligned TeleSleep with VA's preferred practice of patient rather than provider transfers. In the patient transfer model, providers schedule and document in both the provider and patient electronic health records, and bill in the provider's facility. However, reflecting on this period of innovation, TeleSleep team members concluded that the provider transfer model could improve patient safety and care coordination while reducing the administrative burden of frontline clinicians. Further research and development are needed to align the provider transfer model with VA's financial model.

Abstract Varicella zoster virus (VZV) is a highly contagious human herpes virus responsible for causing chickenpox (varicella) and shingles (herpes zoster). Despite the approval of a highly effective vaccine, Shingrix®, the global incidence of herpes zoster is increasing and the economic burden to the health care system and society are substantial due to significant loss of productivity and health complications, particularly among elderly and immunocompromised individuals. This is primarily because access to the vaccines remains mostly limited to countries within developed economies, such as USA and Canada. Therefore, similarly effective vaccines against VZV that are more accessible to the rest-of-the-world are necessary. In this study, we aimed to evaluate immunogenicity and memory response induced by three mRNA-LNP-based vaccine candidates targeting VZV’s surface glycoprotein E (gE). C57BL/6 mice were immunized with each candidate vaccine, and humoral and cellular immune responses were assessed. Our results demonstrate that the mRNA-LNP-based vaccine candidates elicited robust and durable humoral responses specific to the gE antigen. Notably, mice vaccinated with the mRNA-LNP vaccines exhibited significantly higher antigen-specific T-cell cytokine production compared to the group receiving Shingrix®, the current standard of care vaccine. Additionally, mRNA-LNP vaccines induced long-lasting memory response, as evidenced by detection of persistent gE-specific Long-Lived Plasma Cells (LLPCs) and memory T cells four months after final immunization. These findings underscore the potential of our mRNA-LNP-based vaccine candidates in generating potent immune responses against VZV, offering promising prospects for their clinical development as an effective prophylactic vaccine against herpes zoster.

Data Trusts are an important emerging approach to enabling the much wider sharing of data from many different sources and for many different purposes, backed by the confidence of clear and unambiguous data governance. Data Trusts combine the technical infrastructure for sharing data with the governance framework of a legal trust. The concept of a data Trust applied specifically to spatial data offers significant opportunities for new and future applications, addressing some longstanding barriers to data sharing, such as location privacy and data sovereignty. This paper introduces and explores the concept of a ‘spatial data Trust’ by identifying and explaining the key functions and characteristics required to underpin a data Trust for spatial data. The work identifies five key features of spatial data Trusts that demand specific attention and connects these features to a history of relevant work in the field, including spatial data infrastructures (SDIs), location privacy, and spatial data quality. The conclusions identify several key strands of research for the future development of this rapidly emerging framework for spatial data sharing.

Recent work reveals a left digit effect in number line estimation such that adults' and children's estimates for three-digit numbers with different hundreds-place digits but nearly identical magnitudes are systematically different (e.g., 398 is placed too far to the left of 401 on a 0-1000 line, despite their almost indistinguishable magnitudes; Lai et al., 2018, https://doi.org/10.1111/desc.12657). In two preregistered studies (N = 218), we investigate the scope and malleability of the left digit effect. Experiment 1 used a typical forward-oriented 0-1000 number line estimation task and an atypical reverse-oriented 1000-0 number line estimation task. Experiment 2 used the same forward-oriented typical 0-1000 number line estimation task from Experiment 1, but with trial-by-trial corrective feedback. We observed a large left digit effect, regardless of the orientation of the line in Experiment 1 or the presence of corrective feedback in Experiment 2. Further, analyses using combined data showed that the pattern was present across most stimuli and participants. These findings demonstrate a left digit effect that is robust and widely observed, and that cannot be easily corrected with simple feedback. We discuss the implications of the findings for understanding sources of the effect and efforts to reduce it.

Abstract TGFβ signaling is associated with non-response to immune checkpoint blockade in patients with advanced cancers, particularly in the immune-excluded phenotype. While previous work demonstrates that converting tumors from excluded to inflamed phenotypes requires attenuation of PD-L1 and TGFβ signaling, the underlying cellular mechanisms remain unclear. Here, we show that TGFβ and PD-L1 restrain intratumoral stem cell-like CD8 T cell (TSCL) expansion and replacement of progenitor-exhausted and dysfunctional CD8 T cells with non-exhausted T effector cells in the EMT6 tumor model in female mice. Upon combined TGFβ/PD-L1 blockade IFNγhi CD8 T effector cells show enhanced motility and accumulate in the tumor. Ensuing IFNγ signaling transforms myeloid, stromal, and tumor niches to yield an immune-supportive ecosystem. Blocking IFNγ abolishes the anti-PD-L1/anti-TGFβ therapy efficacy. Our data suggest that TGFβ works with PD-L1 to prevent TSCL expansion and replacement of exhausted CD8 T cells, thereby maintaining the T cell compartment in a dysfunctional state.

Introduction: Impaired fracture healing, specifically non-union, has been found to occur up to 14% in tibial shaft fractures. The current standard of care to treat non-union often requires additional surgeries which can result in long recovery times. Injectable-based therapies to accelerate fracture healing have the potential to mitigate the need for additional surgeries. Gene therapies have recently undergone significant advancements due to developments in nanotechnology, which improve mRNA stability while reducing immunogenicity.Methods: In this study, we tested the efficacy of mineral coated microparticles (MCM) and fluoride-doped MCM (FMCM) to effectively deliver firefly luciferase (FLuc) mRNA lipoplexes (LPX) to the fracture site. Here, adult mice underwent a tibia fracture and stabilization method and all treatments were locally injected into the fracture. Level of osteogenesis and amount of bone formation were assessed using gene expression and histomorphometry respectively. Localized and systemic inflammation were measured through gene expression, histopathology scoring and measuring C-reactive protein (CRP) in the serum. Lastly, daily IVIS images were taken to track and measure transfection over time.Results: MCM-LPX-FLuc and FMCM-LPX-FLuc were not found to cause any cytotoxic effects when tested in vitro. When measuring the osteogenic potential of each mineral composition, FMCM-LPX-FLuc trended higher in osteogenic markers through qRT-PCR than the other groups tested in a murine fracture and stabilization model. Despite FMCM-LPX-FLuc showing slightly elevated il-1β and il-4 levels in the fracture callus, inflammation scoring of the fracture callus did not result in any differences. Additionally, an acute systemic inflammatory response was not observed in any of the samples tested. The concentration of MCM-LPX-FLuc and FMCM-LPX-FLuc that was used in the murine fracture model did not stimulate bone when analyzed through stereological principles. Transfection efficacy and kinetics of delivery platforms revealed that FMCM-LPX-FLuc prolongs the luciferase signal both in vitro and in vivo.Discussion: These data together reveal that FMCM-LPX-FLuc could serve as a promising mRNA delivery platform for fracture healing applications.

BackgroundIn fiscal year 2021, the Veterans Health Administration (VHA) provided care for sleep disorders to 599,966 Veterans, including 189,932 rural Veterans. To further improve rural access, the VA Office of Rural Health developed the TeleSleep Enterprise-Wide Initiative (EWI). TeleSleep's telemedicine strategies include tests for sleep apnea at the Veteran's home rather than in a sleep lab; Clinical Video Telehealth applications; and other forms of virtual care. In 2017 and 2020, VHA provided 3-year start-up funding to launch new TeleSleep programs at rural-serving VA medical facilities.MethodsIn early 2022, we surveyed leaders of 24 sites that received TeleSleep funding to identify successes, failures, facilitators, and barriers relevant to sustaining TeleSleep implementations upon expiration of startup funding. We tabulated frequencies on the multiple choice questions in the survey, and, using the survey's critical incident framework, summarized the responses to open-ended questions. TeleSleep program leaders discussed the responses and synthesized recommendations for improvement.Results18 sites reported sustainment, while six were “on track.” Sustainment involved medical centers or regional entities incorporating TeleSleep into their budgets. Facilitators included: demonstrating value; aligning with local priorities; and collaborating with spoke sites serving rural Veterans. Barriers included: misalignment with local priorities; and hiring delays. COVID was a facilitator, as it stimulated adoption of telehealth practices; and also a barrier, as it consumed attention and resources. Recommendations included: longer startup funding; dedicated funding for human resources to accelerate hiring; funders communicating with local facility leaders regarding how TeleSleep aligns with organizational priorities; hiring into job classifications aligned with market pay; and obtaining, from finance departments, projections and outcomes for the return on investment in TeleSleep.

Abstract Burkholderia have potential as biocontrol agents because they encode diverse biosynthetic gene clusters (BGCs) for a range of antimicrobial metabolites. Given the opportunistic pathogenicity associated with Burkholderia species, heterologous BGC expression within non‐pathogenic hosts is a strategy to construct safe biocontrol strains. We constructed a yeast‐adapted Burkholderia‐Escherichia shuttle vector (pMLBAD_yeast) with a yeast replication origin 2 μ and URA3 selection marker and optimised it for cloning BGCs using the in vivo recombination ability of Saccharomyces cerevisiae. Two Burkholderia polyyne BGCs, cepacin (13 kb) and caryoynencin (11 kb), were PCR‐amplified as three overlapping fragments, cloned downstream of the pBAD arabinose promoter in pMLBAD_yeast and mobilised into Burkholderia and Paraburkholderia heterologous hosts. Paraburkholderia phytofirmans carrying the heterologous polyyne constructs displayed in vitro bioactivity against a variety of fungal and bacterial plant pathogens similar to the native polyyne producers. Thirteen Paraburkholderia strains with preferential growth at 30°C compared with 37°C were also identified, and four of these were amenable to genetic manipulation and heterologous expression of the caryoynencin construct. The cloning and successful heterologous expression of Burkholderia biosynthetic gene clusters within Paraburkholderia with restricted growth at 37°C opens avenues for engineering non‐pathogenic biocontrol strains.

Arts-based training programs can aid in building observational skills, communication, team building, and leadership through targeted analysis of works of art. Furthermore, arts-based training programs are unique in that they highlight non-technical skills (NTS). The goal of this study was to investigate the extent to which NTS were developed through an arts-based training program among entry-level occupational therapy doctorate (OTD) students. This study used a mixed methodological approach to analyze retrospective data. Quantitative analysis of the Kentucky Inventory of Mindfulness Skills (KIMS) assessment was conducted. Qualitative analysis was completed through analysis of the participants’ reflection papers. Quantitative results (n = 69) from the KIMS show significance (p = 0.011; 0.013), and qualitative analysis (n = 32) resulted in five central themes: evolution of thinking, holistic approach, diverse perspective, lived reality of context, and development of NTS (collaboration, self-awareness, mindful of others, creativity, and effective communication). Quantitative and qualitative findings suggest arts-based training programs can improve students’ ability to reduce their bias through implementation of a mindfulness lens. Results and recommendations from this study can be used as an example for occupational therapy academic programs to implement an evidence-based teaching methodology.

Abstract Theoretical evolution within the field of Open Pedagogy has progressed rapidly in recent years. Practical application, however, has lagged this conceptual development. This article seeks to explore the gap between theory and practice by examining how the use of terms within the open education space may create barriers for instructors new to the concept. The authors of this article argue that the most effective approach to creating momentum toward practices associated with Open Pedagogy is to begin with an alignment of an instructor’s values with the attributes of an open educator. Subject matter experts were consulted in the association of open pedagogical values with specific practices. The result is a visual aid useful for an instructor’s self-assessment or in collaboration with curriculum designers to identify a logical start point for an instructor as they begin their movement from a more traditional to an open approach.

Abstract Maleidrides are a family of structurally related fungal natural products, many of which possess diverse, potent bioactivities. Previous identification of several maleidride biosynthetic gene clusters, and subsequent experimental work, has determined the ‘core’ set of genes required to construct the characteristic medium-sized alicyclic ring with maleic anhydride moieties. Through genome mining, this work has used these core genes to discover ten entirely novel putative maleidride biosynthetic gene clusters, amongst both publicly available genomes, and encoded within the genome of the previously un-sequenced epiheveadride producer Wicklowia aquatica CBS 125634. We have undertaken phylogenetic analyses and comparative bioinformatics on all known and putative maleidride biosynthetic gene clusters to gain further insights regarding these unique biosynthetic pathways.

Abstract The rise in throughput and quality of long-read sequencing should allow unambiguous identification of full-length transcript isoforms. However, its application to single-cell RNA-seq has been limited by throughput and expense. Here we develop and characterize long-read Split-seq (LR-Split-seq), which uses combinatorial barcoding to sequence single cells with long reads. Applied to the C2C12 myogenic system, LR-split-seq associates isoforms to cell types with relative economy and design flexibility. We find widespread evidence of changing isoform expression during differentiation including alternative transcription start sites (TSS) and/or alternative internal exon usage. LR-Split-seq provides an affordable method for identifying cluster-specific isoforms in single cells.

[This corrects the article DOI: 10.1371/journal.pntd.0008604.].

Students acting as co-creators of academic material is growing in popularity as a pedagogical approach in higher education. With student engagement and persistence consistently being emphasized for student and institution well-being, educational praxis must foster engaged, high-retention student cohorts. This exploratory research uses a mixed-methods approach to examine the experience of students participating in a first-year course utilizing OER-enabled Pedagogy. Students considered how projects that were open impacted their perception of course engagement, satisfaction, and overall experience. Participants also evaluated their level of concern in sharing attributed academic work. A plurality of students preferred the project using OER-enabled Pedagogy, indicating it increased engagement and skills acquisition. The majority of students were unconcerned about sharing work publicly, even if their names were included. Themes that emerged from interviews included the motivational value of creating work potentially valuable to others, being given agency, and receiving public credit for their efforts.

Abstract Background Obstructive sleep apnea is common among rural Veterans, however, access to diagnostic sleep testing, sleep specialists, and treatment devices is limited. To improve access to sleep care, the Veterans Health Administration (VA) implemented a national sleep telemedicine program. The TeleSleep program components included: 1) virtual clinical encounters; 2) home sleep apnea testing; and 3) web application for Veterans and providers to remotely monitor symptoms, sleep quality and use of positive airway pressure (PAP) therapy. This study aimed to identify factors impacting Veteran’s participation, satisfaction and experience with the TeleSleep program as part of a quality improvement initiative. Methods Semi-structured interview questions elicited patient perspectives and preferences regarding accessing and engaging with TeleSleep care. Rapid qualitative and matrix analysis methods for health services research were used to organize and describe the qualitative data. Results Thirty Veterans with obstructive sleep apnea (OSA) recruited from 6 VA telehealth “hubs” participated in interviews. Veterans reported positive experiences with sleep telemedicine, including improvements in sleep quality, other health conditions, and quality of life. Access to care improved as a result of decreased travel burden and ability of both clinicians and Veterans to remotely monitor and track personal sleep data. Overall experiences with telehealth technology were positive. Veterans indicated a strong preference for VA over non-VA community-based sleep care. Patient recommendations for change included improving scheduling, continuity and timeliness of communication, and the equipment refill process. Conclusions The VA TeleSleep program improved patient experiences across multiple aspects of care including a reduction in travel burden, increased access to clinicians and remote monitoring, and patient-reported health and quality of life outcomes, though some communication and continuity challenges remain. Implementing telehealth services may also improve the experiences of patients served by other subspecialties or healthcare systems.

Number line estimation (NLE) performance is usually believed to depend on the magnitudes of presented numerals, rather than on the particular digits instantiating those magnitudes. Recent research, however, shows that NLE placements differ considerably for target numerals with nearly identical magnitudes, but instantiated with different leftmost digits. Here we investigate whether this left digit effect may be due, in part, to the ordering of digits in number words. In English, the leftmost digit of an Arabic numeral is spoken first (“forty-one”), but Dutch number words are characterized by the inversion property: the rightmost digit of a two-digit number word is spoken first (“eenenveertig” – one and forty in Dutch). Participants (N = 40 Dutch-English bilinguals and N = 20 English-speaking monolinguals) completed a standard 0-100 NLE task. Target numerals were read aloud by an experimenter in either English or Dutch. Preregistered analyses revealed a strong left digit effect in monolingual English speakers’ estimates: e.g., 41 was placed more than two units to the right of 39. No left digit effect was observed among Dutch-English bilingual participants tested in either language. These findings are consistent with the idea that the order in which digits are spoken might influence multi-digit number processing, and suggests linguistic influences on numerical estimation performance.

Single-cell RNA-seq (scRNA-seq) has revolutionized modern genomics, but the large size of myotubes and myofibers has restricted use of scRNA-seq in skeletal muscle. For the study of muscle, single-nucleus RNA-seq (snRNA-seq) has emerged not only as an alternative to scRNA-seq, but as a novel method providing valuable insights into multinucleated cells such as myofibers. Nuclei within myofibers specialize at junctions with other cell types such as motor neurons. Nuclear heterogeneity plays important roles in certain diseases such as muscular dystrophies. We survey current methods of high-throughput single cell and subcellular resolution transcriptomics, including single-cell and single-nucleus RNA-seq and spatial transcriptomics, applied to satellite cells, myoblasts, myotubes and myofibers. We summarize the major myonuclei subtypes identified in homeostatic and regenerating tissue including those specific to fiber type or at junctions with other cell types. Disease-specific nucleus populations were found in two muscular dystrophies, FSHD and Duchenne muscular dystrophy, demonstrating the importance of performing transcriptome studies at the single nucleus level in muscle.

Abstract Background Youth with autism spectrum disorder (ASD) represent a growing population with significant service needs. Prominent among these needs are high rates of co-occurring psychiatric conditions that contribute to increased functional impairments and often necessitate mental health services. Executive functioning deficits are associated with ASD as well as common co-occurring conditions (e.g., attention-deficit/hyperactivity disorder) and an evidence-based intervention has been developed and tested to address executive functioning within the school context. There is an urgent need to implement indicated evidence-based interventions for youth with ASD receiving care in community mental health settings. Interventions that optimally “fit” the mental health services context as well as the complex and co-occurring mental health needs of these youth have the potential to improve key clinical outcomes for this high priority population. Methods This mixed-methods developmental study will apply the Exploration, Preparation, Implementation, Sustainment implementation framework and a community-academic partnership approach to systematically adapt and test an evidence-based executive functioning intervention for youth with ASD for delivery in community mental health settings. Specific aims are to (1) conduct a need and context assessment to inform the systematic adaptation an executive functioning evidence-based intervention; (2) systematically adapt the clinical intervention and develop a corresponding implementation plan, together entitled “Executive Functioning for Enhancing Community-based Treatment for ASD,” (EFFECT for ASD); and (3) conduct a feasibility pilot test of EFFECT for ASD in community mental health settings. Discussion Tailoring evidence-based interventions for delivery in community-based mental health services for youth with ASD has the potential to increase quality of care and improve child outcomes. Results from the current study will serve as the foundation for large-scale hybrid implementation and effectiveness trials and a generalizable approach for different service systems of care and clinical populations. Trial registration Clinicaltrials.gov, NCT04295512 .

The COVID-19 pandemic has forced both quantitative and qualitative health researchers to adapt and strategize data collection strategies without causing any harm to the participants or researchers. This has resulted in utilizing various types of strategies such as online surveys and synchronous virtual platforms such as Zoom and Webex. This transition from face-to-face to synchronous online platforms has helped in increasing coverage as well as reaching participants who are otherwise unreachable. While quantitative health researchers seem to have made a seamless transition to synchronous online platforms, qualitative health researchers who rely on studying participants in their “real-world-settings” are facing unique challenges with online data collection strategies. This article critically examines the benefits and challenges of implementing qualitative health research studies via synchronous online platforms and provides several practical considerations that can inform qualitative health researchers. It can also assist Institutional Review Board members in reviewing and implementing qualitative health research study protocols in a manner that preserves the integrity, richness, and iterative nature of qualitative research methodology.

COVID-19, in addition to disrupting the global education system in general, is widening the economic and racial gaps institutions have spent years trying to address. The economic reality is that students who work to support themselves, their families, and purchase educational materials needed to succeed have been disproportionately harmed. This article discusses how the global COVID-19 pandemic is compounding structural inequities inherent in higher education. This requires faculty to reevaluate their role as agents of change in a world that is fundamentally different than it was a short time ago. The experience of one liberal arts institution in the U.S. who moved all courses to free materials in under six months will be recounted as an example of what is possible during extraordinary circumstances if students are truly prioritized during strategic planning.

Open pedagogy is growing in popularity as an instructional method to decentralize classroom power dynamics, engage students, and provide greater meaning to student work. To investigate the impact of open pedagogy on motivation, interviews were conducted with first-year college students at a four-year liberal arts college after completing a semester-long project based on this pedagogical approach. Student responses were assessed using self-determination theory as a theoretical framework, particularly in relation to the motivation regulatory styles displayed by research participants. Results indicate that students experienced various forms of extrinsic motivation during the project based on open pedagogy, with autonomous forms of regulation being more prevalent than controlled regulation. Interview data also suggest that agency plays a role in mediating the internalization of student motivation. Based on these findings, suggestions are provided to the design of assignments in general and open pedagogy specifically to enhance development of autonomous forms of motivation.

BackgroundSustainability within neglected tropical disease (NTD) programs is a complex and challenging issue. The need for a shared understanding about what sustainability means for NTD programs is more important than ever as stakeholders are currently realigning for the next decade of NTD programming with the launch of WHO's new NTD roadmap for 2012-2030. The aim of this paper is to assess different perspectives to generate a working definition of sustainability for NTD programs.Methodology/principal findingsThis study surveyed affiliates of the NTD NGO Network (NNN) about their definitions of sustainability and then analyzed the data using an inductive and deductive process. The research team drafted a sustainability statement based on the survey findings and then solicited and incorporated feedback on the statement from a diverse group of expert reviewers. The final statement includes a working definition of sustainability for NTD programs that highlights three key essential components to sustainability: domestic commitment, responsive resource mobilization, and accountability.Conclusions/significanceThis research resulted in a sustainability statement, based on a survey and extensive consultation with stakeholders, that represents a starting point for shared understanding around the concept of sustainability for NTD programs. Future collaborative work should build off this definition and seek to incorporate indicators for sustainability into programmatic decision-making.

Students acting as content creators is an emergent trend in the field of open educational practice. As more faculty turn towards the use of open pedagogy or OER-enabled Pedagogy, they must be prepared to address concerns related to intellectual property rights of student work. This article addresses student concerns related to intellectual property rights, specifically related to Creative Commons licensing as well as faculty awareness of use of Creative Commons licensing. Research was conducted at a small, liberal arts college in the Appalachian Region of the United States. All first-year students engaged in an OER-enabled Pedagogy project where they collaboratively created a reader for the First Year Studies seminar course. Following class, students and faculty were interviewed regarding how dynamics of intellectual property and Creative Commons licensing impacted the educational process. Results indicate students are open to sharing their works with credit, and value helping others. Faculty tend to be unfamiliar with Creative Commons licensing and must balance the desire to help students understand licensing and prescribing their own preferences when asked about licensing selection.

Students are experiencing enormous economic precarity as a result of COVID-19. Reports indicate that those hardest hit by job loss due to the coronavirus are of lower income (Beer, 2020). While economic recovery is underway for more affluent workers, the same cannot be said for those toward the bottom of the wage scale, particularly underserved populations (Long, 2020). The University of Pikeville (UPIKE) in Central Appalachia recognizes the impact that emerging and existing financial inequities have on our institution’s most vulnerable populations. Even pre-pandemic, students had indicated the costs of purchasing textbooks was stressful and discouraging. As a result, the institution decided in April 2020 to convert all classes to free materials by the start of the Fall 2020 semester. Written by those who supported faculty in this transition, the goal of this article is threefold: To describe our research methods in seeking out appropriate free material for instructors and how we guided faculty in developing their own knowledge in search strategies, To detail the process we established for faculty to apply for funding when appropriate materials could not be found, and To share the lessons we learned along with emerging success stories. We hope that this guidance will encourage other institutions to implement similar initiatives. Based on anecdotal experiences as well as our own ongoing research, we believe similar efforts are essential in addressing systemic inequities and creating cross-campus collaboration, particularly in the face of unprecedented challenges that were not created by, but have been exacerbated because of the global pandemic.

Abstract Introduction Recent work reveals a new source of error in number line estimation (NLE), the left digit effect (Lai, Zax, et al., 2018), whereby numerals with different leftmost digits but similar magnitudes (e.g., 399, 401) are placed farther apart on a number line (e.g., 0 to 1,000) than is warranted. The goals of the present study were to: (1) replicate the left digit effect, and (2) assess whether it is related to mathematical achievement. Method Participants were all individuals (adult college students) who completed the NLE task in the laboratory between 2014 and 2019 for whom SAT scores were available (n = 227). Results We replicated the left digit effect but found its size was not correlated with SAT math score, although it was negatively correlated with SAT verbal score for one NLE task version. Conclusions These findings provide further evidence that individual digits strongly influence estimation performance and suggest that this effect may have different cognitive contributors, and predict different complex skills, than overall NLE accuracy.

To aid students during COVID-19, the University of Pikeville transitioned all undergraduate classes to no-cost alternatives for course textbooks. Additionally, the academic calendar was modified for the first time in the institution’s 130-year history from a traditional 16-week semester to 8-week block scheduling. This case study explores strategies, approaches to corresponding with constituents, and lessons learned in leading an institution through two major curricular changes in under six months in response to a global pandemic.

BackgroundIn Kenya, over five million school age children (SAC) are estimated to be at risk of parasitic worms causing soil-transmitted helminthiasis (STH) and schistosomiasis. As such, the Government of Kenya launched a National School Based Deworming (NSBD) program in 2012 targeting the at-risk SAC living in endemic regions, with the aim of reducing infections prevalence to a level where they no longer constitute a public health problem. The impact of the program has been consistently monitored from 2012 to 2017 through a robust and extensive monitoring and evaluation (M&E) program. The aim of the current study was to evaluate the parasitological outcomes and additionally investigate water, sanitation and hygiene (WASH) related factors associated with infection prevalence after five rounds of mass drug administration (MDA), to inform the program's next steps.Materials and methodsWe utilized a cross-sectional design in a representative, stratified, two-stage sample of school children across six regions in Kenya. A sample size of 100 schools with approximately 108 children per school was purposively selected based on the Year 5 STH infection endemicity prior to the survey. Stool samples were examined for the presence of STH and Schistosoma mansoni eggs using double-slide Kato-Katz technique, urine samples were processed using urine filtration technique for the presence of S. haematobium eggs. Survey questionnaires were administered to all the participating children to collect information on their demographic and individual, household and school level WASH characteristics.Principal findingsOverall, STH prevalence was 12.9% (95%CI: 10.4-16.1) with species prevalence of 9.7% (95%CI: 7.5-12.6) for Ascaris lumbricoides, 3.6% (95%CI: 2.2-5.8) for Trichuris trichiura and 1.0% (95%CI: 0.6-1.5) for hookworm. S. mansoni prevalence was 2.2% (95%CI: 1.2-4.3) and S. haematobium prevalence was 0.3% (95%CI: 0.1-1.0). All the infections showed significant prevalence reductions when compared with the baseline prevalence, except S. mansoni. From multivariable analysis, increased odds of any STH infections were associated with not wearing shoes, adjusted odds ratio (aOR) = 1.36 (95%CI: 1.09-1.69); p = 0.007; high number of household members, aOR = 1.21 (95%CI: 1.04-1.41); p = 0.015; and school absenteeism of more than two days, aOR = 1.33 (95%CI: 1.01-1.80); p = 0.045. Further, children below five years had up to four times higher odds of getting STH infections, aOR = 4.68 (95%CI: 1.49-14.73); p = 0.008. However, no significant factors were identified for schistosomiasis, probably due to low prevalence levels affecting performance of statistical analysis.ConclusionsAfter five rounds of MDA, the program shows low prevalence of STH and schistosomiasis, however, not to a level where the infections are not a public health problem. With considerable inter-county infection prevalence heterogeneity, the program should adopt future MDA frequencies based on the county's infection prevalence status. Further, the program should encourage interventions aimed at improving coverage among preschool age children and improving WASH practices as long-term infection control strategies.

FSHD is characterized by the misexpression of DUX4 in skeletal muscle. Although DUX4 upregulation is thought to be the pathogenic cause of FSHD, DUX4 is lowly expressed in patient samples, and analysis of the consequences of DUX4 expression has largely relied on artificial overexpression. To better understand the native expression profile of DUX4 and its targets, we performed bulk RNA-seq on a 6-day differentiation time-course in primary FSHD2 patient myoblasts. We identify a set of 54 genes upregulated in FSHD2 cells, termed FSHD-induced genes. Using single-cell and single-nucleus RNA-seq on myoblasts and differentiated myotubes, respectively, we captured, for the first time, DUX4 expressed at the single-nucleus level in a native state. We identified two populations of FSHD myotube nuclei based on low or high enrichment of DUX4 and FSHD-induced genes ("FSHD-Lo" and "FSHD Hi", respectively). FSHD-Hi myotube nuclei coexpress multiple DUX4 target genes including DUXA, LEUTX and ZSCAN4, and also upregulate cell cycle-related genes with significant enrichment of E2F target genes and p53 signaling activation. We found more FSHD-Hi nuclei than DUX4-positive nuclei, and confirmed with in situ RNA/protein detection that DUX4 transcribed in only one or two nuclei is sufficient for DUX4 protein to activate target genes across multiple nuclei within the same myotube. DUXA (the DUX4 paralog) is more widely expressed than DUX4, and depletion of DUXA suppressed the expression of LEUTX and ZSCAN4 in late, but not early, differentiation. The results suggest that the DUXA can take over the role of DUX4 to maintain target gene expression. These results provide a possible explanation as to why it is easier to detect DUX4 target genes than DUX4 itself in patient cells and raise the possibility of a self-sustaining network of gene dysregulation triggered by the limited DUX4 expression.

When allocating resources, people often diversify across categories even when those categories are arbitrary, such that allocations differ when identical sets of options are partitioned differently ("partition dependence"). The first goal of the present work (Experiment 1) was to replicate an experiment by Fox and colleagues in which graduate students exhibited partition dependence when asked how university financial aid should be allocated across arbitrarily partitioned income brackets. Our sample consisted of community members at a liberal arts college where financial aid practices have been recent topics of debate. Because stronger intrinsic preferences can reduce partition dependence, these participants might display little partition dependence with financial aid allocations. Alternatively, a demonstration of strong partition dependence in this population would emphasize the robustness of the effect. The second goal was to extend a "high transparency" modification to the present task context (Experiment 2) in which participants were shown both possible income partitions and randomly assigned themselves to one, to determine whether partition dependence in this paradigm would be reduced by revealing the study design (and the arbitrariness of income categories). Participants demonstrated clear partition dependence in both experiments. Results demonstrate the robustness of partition dependence in this context.

Toxicology studies on pristine graphene are limited and lack significant correlations with actual human response. The goal of the current study was to determine the response of total colonic human tissue to pristine graphene exposure. Biopsy punches of colon tissues from healthy human were used to assess the biological response after ex vivo exposure to graphene at three different concentrations (1, 10, and 100 µg/mL). mRNA expression of specific genes or intestinal cytokine abundance was assessed using real-time PCR or multiplex immunoassays, respectively. Pristine graphene-activated genes that are related to binding and adhesion (GTPase and KRAS) within 2 h of exposure. Furthermore, the PCNA (proliferating cell nuclear antigen) gene was upregulated after exposure to graphene at all concentrations. Ingenuity pathway analysis revealed that STAT3 and VEGF signaling pathways (known to be involved in cell proliferation and growth) were upregulated. Graphene exposure (10 µg/mL) for 24 h significantly increased levels of pro-inflammatory cytokines IFNγ, IL-8, IL-17, IL-6, IL-9, MIP-1α, and Eotaxin. Collectively, these results indicated that graphene may activate the STAT3–IL23–IL17 response axis. The findings in this study provide information on toxicity evaluation using a human-relevant ex vivo colon model and serve as a basis for further exploration of its bio-applications.

Limited antibacterial activity of silver ions leached from silver-impregnated food contact materials could be due to: 1) the presence of silver resistance genes in tested bacteria; or 2) lack of susceptibility to silver ion-mediated killing in the bacterial strain (K. Williams, L. Valencia, K. Gokulan, R. Trbojevich, S. Khare, 2016 [1]). This study contains data to address the specificity of silver resistance genes in Salmonella Typhimurium during the real time PCR using melting curve analysis and an assessment of the minimum inhibitory concentration of silver ions for Salmonella.

Consumer products manufactured with antimicrobial silver nanoparticles (AgNPs) may affect the gastrointestinal (GI) system. The human GI-tract is complex and there are physiological and anatomical differences between human and animal models that limit comparisons between species. Thus, assessment of AgNP toxicity on the human GI-tract may require tools that allow for the examination of subtle changes in inflammatory markers and indicators of epithelial perturbation. Fresh tissues were excised from the GI-tract of human male and female subjects to evaluate the effects of AgNPs on the GI-system. The purpose of this study was to perform an assessment on the ability of the ex vivo model to evaluate changes in levels of pro-/anti-inflammatory cytokines/chemokines and mRNA expression of intestinal permeability related genes induced by AgNPs in ileal tissues. The ex vivo model preserved the structural and biological functions of the in-situ organ. Analysis of cytokine expression data indicated that intestinal tissue of male and female subjects responded differently to AgNP treatment, with male samples showing significantly elevated Granulocyte-macrophage colony-stimulating factor (GM-CSF) after treatment with 10 nm and 20 nm AgNPs for 2 h and significantly elevated RANTES after treatment with 20 nm AgNPs for 24 h. In contrast, tissues of female showed no significant effects of AgNP treatment at 2 h and significantly decreased RANTES (20 nm), TNF-α (10 nm), and IFN-γ (10 nm) at 24 h. Smaller size AgNPs (10 nm) perturbed more permeability-related genes in samples of male subjects, than in samples from female subjects. In contrast, exposure to 20 nm AgNPs resulted in upregulation of a greater number of genes in female-derived samples (36 genes) than in male-derived samples (8 genes). The ex vivo tissue model can distinguish sex dependent effects of AgNP and could serve as a translational non-animal model to assess the impacts of xenobiotics on human intestinal mucosa.

Purpose: This report describes a case of amaurosis fugax due to a rare primary cardiac sarcoma. Observations: A patient who was recently diagnosed with left atrial pleomorphic sarcoma presented with a chief complaint of multiple episodes of intermittent vision loss in the right eye during the course of radiation therapy. Conclusions and importance: The authors postulate emboli from the left atrial sarcoma entered systemic circulation and subsequently caused brief episodes of transient occlusion to retinal, ophthalmic and/or ciliary arteries leading to momentary retinal hypoxia. We believe this is a novel finding, previously unreported in the literature, of transient embolic occlusion without permanent visual sequelae due to a malignant primary cardiac pleomorphic sarcoma.

Most nematodes are small worms that lack enough RNA for regular RNA-seq protocols without pooling hundred to thousand of individuals. We have adapted the Smart-seq2 protocol in order to sequence the transcriptome of an individual worm. While developed for individual Steinernema carpocapsae and Caenorhabditis elegans larvae as well as embryos, the protocol should be adaptable for other nematode species and small invertebrates. In addition, we describe how to analyze the RNA-seq results using the Galaxy online environment. We expect that this method will be useful for the studying gene expression variances of individual nematodes in wild type and mutant backgrounds.

While there is substantial evidence that adults who violate gender stereotypes often face backlash (i.e. social and economic penalties), less is known about the nature of gender stereotypes for young children, and the penalties that children may face for violating them. We conducted three experiments, with over 2000 adults from the US, to better understand the content and consequences of adults' gender stereotypes for young children. In Experiment 1, we tested which characteristics adults (N = 635) believed to be descriptive (i.e. typical), prescriptive (i.e. required), and proscriptive (i.e. forbidden) for preschool-aged boys and girls. Using the characteristics that were rated in Experiment 1, we then constructed vignettes that were either 'masculine' or 'feminine', and manipulated whether the vignettes were said to describe a boy or a girl. Experiment 2 (N = 697) revealed that adults rated stereotype-violating children as less likeable than their stereotype-conforming peers, and that this difference was more robust for boys than girls. Experiment 3 (N = 731) was a direct replication of Experiment 2, and revealed converging evidence of backlash against stereotype-violating children. In sum, our results suggest that even young children encounter backlash from adults for stereotype violations, and that these effects may be strongest for boys.

Mutations in the actin-binding gene Filamin-A have been linked to non-syndromic myxomatous valvular dystrophy and associated mitral valve prolapse. Previous studies by our group traced the adult valve defects back to developmental errors in valve interstitial cell-mediated extracellular matrix remodeling during fetal valve gestation. Mice deficient in Filamin-A exhibit enlarged mitral leaflets at E17.5, and subsequent progression to a myxomatous phenotype is observed by two months. For this study, we sought to define mechanisms that contribute to myxomatous degeneration in the adult Filamin-A-deficient mouse. In vivo experiments demonstrate increased infiltration of hematopoietic-derived cells and macrophages in adolescent Filamin-A conditional knockout mice. Concurrent with this infiltration of hematopoietic cells, we show an increase in Erk activity, which localizes to regions of MMP2 expression. Additionally, increases in cell proliferation are observed at two months, when hematopoietic cell engraftment and signaling are pronounced. Similar changes are observed in human myxomatous mitral valve tissue, suggesting that infiltration of hematopoietic-derived cells and/or increased Erk signaling may contribute to myxomatous valvular dystrophy. Consequently, immune cell targeting and/or suppression of pErk activities may represent an effective therapeutic option for mitral valve prolapse patients.

The study of perinatal mental health (mental health during pregnancy and postpartum) is a complex field of study that is of major importance both for the mental and physical health of new mothers and for the neurobehavioral development and long-term functioning of the children they bear. In this review, we cover the most recent additions to this rapidly evolving field. Notable advances include further illumination of the epidemiological patterns and clinical manifestations of perinatal mood disruption; new efficacy data on treatment and prevention; clarifications of the respective contributions of maternal mental illness and psychotropic medication to outcomes of pregnancy, birth, and child development; and updated expert guidelines for screening.

Introduction. Our study aims to investigate and evaluate (1) rates of success of ECV for breech presentation at term at the Royal Women’s Hospital in comparison to international standards; (2) mode of delivery following ECV; (3) factors influencing success rates of ECV at the Royal Women’s Hospital. Methods. An audit of all women who underwent ECV between the years 2007 and 2014 at the Royal Women’s Hospital as public patients was completed. Data parameters were collected from paper and electronic patient files at the Women’s Hospital. Data was collected to analyse the effect of the following parameters on ECV success and birth outcome: age, parity, gestational age, BMI, AFI, and tocolytic use. These parameters were analysed to determine their effect on ECV outcome and birth outcome. Results. The Women’s Hospital, Melbourne, has an ECV success rate of approximately 37%. Of the patients undergoing ECV, 29% proceeded to normal vaginal delivery. Predictors of successful ECV included low BMI, multiparity, and AFI more than 16 (P

Desde o início de sua carreira como compositor e líder de orquestra de jazz nos anos de 1920, Duke Ellington tem sido comparado a compositores eruditos europeus como Delius e Debussy. Declarações como a de Constant LAMBERT (1934), de que Duke Ellington estabeleceu "um padrão pelo qual podemos julgar. . .compositores eruditos" se concentraram em procedimentos composicionais complexos de sua obra. Seguindo outra direção, o presente estudo examina a interseção entre os estilos erudito e jazzístico, por meio da análise de seções tipicamente improvisadas de Ellington. O desenvolvimento do solo improvisado de saxofone barítono, materiais improvisatórios no Interlúdio e o papel do piano de Duke Ellington em três gravações - de 1937, 1953 e 1956 (ELLINGTON, 1937, 1953, 1956), - de Diminuendo and crescendo in blue (1937) indicam o estabelecimento de solos fixos no repertório de sua orquestra. O grau de elaboração composicional aí implícito sugere outras implicações. A análise minuciosa das gravações e a verificação de críticas contemporâneas de Ellington e de fontes acadêmicas posteriores (especialmente a de Bruno NETTL, 1974) permitem avaliar as implicações da predeterminação sugerida pelo tratamento que Duke Ellington deu à improvisação neste período.