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To design, implement, and evaluate a digital indoor wayfinding web application (KH Wayfinder) for a tertiary care hospital, assessing its effects on spatial orientation and navigation-related stress among visitors. A 3-phase study was conducted in a tertiary care hospital in coastal Karnataka, India, from April 1, 2023, through July 31, 2024. Phase 1 involved a cross-sectional survey (n=41) to assess user attitudes toward digital wayfinding. In phase 2, a browser-based application was developed using HyperText Markup Language, JavaScript, cascading style sheets, and Leaflet.js, covering 5 hospital floors with 52 destination points and 758 routes. Phase 3 consisted of usability testing with 54 participants using a validated questionnaire to assess performance, satisfaction, and ease of use. The majority of users 33 (80.5%) expressed willingness to use a digital Wayfinder. Postimplementation results showed that 46 (85.2%) found the tool easy to use, 47 (87%) reported a reduction in navigation time, and 45 (83.3%) experienced reduced psychological stress. Additionally, 51 (94.4%) preferred the digital system over traditional signage, and 54 (100%) would recommend it to others. KH Wayfinder demonstrated high usability, effectiveness, and user satisfaction as a low-cost digital navigation solution. Its browser-based architecture and open-source design make it scalable and adaptable for broader use in smart hospital environments. Future enhancements may include real-time positioning, multilingual support, and accessibility features.

MYC amplification contributes to poor survival and outcome in pancreatic ductal adenocarcinoma (PDAC). Here we show that in PDAC cell lines with amplified MYC, MondoA is required for viability, facilitating proliferation while suppressing apoptosis in vitro and in vivo. Transcriptional and genomic profiling demonstrates that loss of MondoA leads to altered expression of direct MondoA targets as well as MYC target genes and is accompanied by shifts in genomic occupancy of MYC, MNT, and the MondoA paralog ChREBP. This altered genomic binding by MYC network members is associated with transcriptional perturbation of multiple metabolic and stress pathways, as well as global changes in N6-methyladenosine modification (m6A) of messenger RNA (mRNA). MondoA inhibition disrupts coordination between MYC network members and the Integrated Stress Response (ISR), resulting in decreased translation of ATF4 mRNA, discordant gene regulation of shared targets of MYC and ATF4 and, ultimately, apoptosis. Reestablishing ATF4 protein expression rescues the diminished viability due to loss of MondoA expression or activity, providing direct evidence of a link between deregulated MYC and the transcriptional machinery of the ISR. Last, we find that small-molecule inhibition of MondoA is lethal in a subset of PDAC cell lines, including patient-derived organoids, suggesting that the ability to target MYC via chemical inhibition of MondoA transcriptional activity may have broad efficacy.

Myotonic dystrophy type 1 (DM1) is a slowly progressive, multi-systemic disorder with clinical phenotypes that vary by age of onset and severity of symptoms. It is the most common form of muscular dystrophy occurring in adults. Abnormal regulation of alternative splicing of pre-mRNA results from a repeat expansion mutation in the dystrophia myotonica protein kinase (DMPK) gene. The resulting spliceopathy is universal across affected individuals and clinical phenotypes and drives the clinical manifestations of DM1, which include a diverse array of signs and symptoms affecting most organ systems. There is currently no disease-modifying treatment for DM1. Heterogeneity in the developmental and degenerative features and patterns of DM1 complicates the stratification, powering, and execution of interventional clinical trials in a reasonable timeframe. The use of splicing change as a surrogate endpoint in DM1 evolved from the principle that the degree of DM1-affected exons relates to the level of functional muscleblind-like (MBNL) RNA-binding protein activity in muscle cell nuclei. Surrogate endpoints based on panels of mis-splicing events reflecting the underlying DM1 molecular mechanism are reasonably likely to predict clinical benefit in a timely fashion, thus enabling accelerated clinical development of therapies that address unmet needs in DM1. Natural history data from the DM1 population support a strong correlation between dysregulated splicing and muscle function and point to the utility of a composite splicing index as a surrogate endpoint to predict future functional benefit, particularly in clinical trials of reasonable duration. Ongoing and future clinical trials will hopefully address the validity of surrogate endpoints using changes in splicing and whether the correction of spliceopathy correlates with meaningful clinical outcome assessments in individuals with DM1.

Strategies to Embrace Living with Lupus Fearlessly (SELF) is an online self-management education programme for people with SLE. This mixed-methods study examines SELF's impact on patient-reported outcomes while assessing implementation outcomes informing feasibility and dissemination. A convergent mixed-methods design was used, merging qualitative and quantitative data to enhance interpretation. The Reach, Effectiveness, Adoption, Implementation and Maintenance evaluation framework was used to evaluate programme feasibility and sustainment potential. Participants were recruited from the Georgians Organized Against Lupus (GOAL) cohort (Georgia). 221 adults with SLE from the GOAL cohort enrolled in SELF, completing patient-reported assessments at baseline and 90-day follow-up. 12 participants also completed in-depth interviews exploring the impact of the programme. Certain subgroups including black participants (n=193; 95% CI=-1.59 to -0.04), those with high fatigue levels (n=164; 95% CI=-1.85 to -0.11) and participants living below 100% of the federal poverty level (n=74; 95% CI=-3.44 to -0.18) reported significantly lower disease activity at 90-day follow-up after engaging with SELF. High-fatigue participants improved on multiple measures while the low-fatigue group showed unexpected declines that may reflect baseline confounding. Qualitative findings generally supported quantitative results. One exception was self-efficacy in managing medications, which showed a small but significant decrease (mean reduction=-1.38, 95% CI=-2.50 to -0.26, Cohen's d=-0.14). However, qualitative data suggested participants became more aware of skill gaps rather than less capable. SELF showed promising results for reducing disease activity, pain and fatigue among specific subgroups of participants with SLE. Further research should broaden the evaluation of SELF to new geographical settings, broader populations (such as people living with lupus in rural healthcare settings) and further testing for cultural relevance across diverse racial and ethnic groups.

Bayesian Metabolic Control Analysis (BMCA) is a promising framework for inferring metabolic control coefficients in data-limited scenarios, combining Bayesian inference with linear-logarithmic (lin-log) rate laws. These metabolic control coefficients quantify how changes in enzyme activities affect steady-state fluxes and metabolite concentrations across a metabolic network. However, its predictive accuracy and limitations remain underexplored. This study systematically evaluates BMCA's ability to infer elasticity values, flux control coefficients (FCC), and concentration control coefficients (CCC) under varying data availability conditions using three synthetic metabolic network models. We demonstrate that BMCA predictions are highly dependent on the inclusion of flux and enzyme concentration data, with the omission of these datasets leading to severe inaccuracies. In our synthetic, enzyme-perturbation datasets, external metabolite concentrations had minimal impact and, in some cases, their exclusion improved predictions; when external-nutrient perturbations were introduced and those concentrations were observed, gains were at most modest. Additionally, we find that posterior estimation with both ADVI and HMC can underestimate large-magnitude elasticities in our synthetic settings, with ADVI showing somewhat higher variance under strong up-regulation; thus, recovering |elasticity| [Formula: see text]1.5 remains challenging regardless of the inference engine. ADVI also fails to accurately infer allosteric interactions, even when regulatory effects are strong. While BMCA maintains reasonable accuracy in partially recovering the rankings of the highest FCC values, its estimates of absolute values remain constrained by prior assumptions and data limitations. Our findings reveal the BMCA algorithm's strengths and weaknesses, providing guidance on its application in metabolic engineering, and highlighting the need for methodological refinements to enhance its predictive capabilities.

The Micrapis subgenus, which includes the black dwarf honey bee (Apis andreniformis) and the red dwarf honey bee (Apis florea), remains underrepresented in genomic studies despite its ecological significance. Here, we present high-quality de novo genome assemblies for both species, generated using a hybrid sequencing approach combining Oxford Nanopore Technologies long reads with Illumina short reads. The final assemblies are highly contiguous, with contig N50 values of 5.0 Mb (A. andreniformis) and 4.3 Mb (A. florea), representing a major improvement over the previously published A. florea genome. Genome completeness assessments indicate high quality, with BUSCO scores exceeding 98.5% using the Hymenoptera database and k-mer analyses supporting base-level accuracy. Repeat annotation revealed a relatively low repetitive sequence content (∼6%), consistent with other Apis species. Using RNA sequencing data, we annotated 12,189 genes for A. andreniformis and 12,207 genes for A. florea, with ∼98% completeness in predicted proteomes. These genome assemblies provide a valuable resource for comparative and functional genomic studies, with the potential to offer new insights into the genetic basis of dwarf honey bee adaptations.

Myotonic dystrophy type 1 (DM1) is a complex multisystemic disease caused by a CTG repeat expansion in the DMPK gene for which there are no approved disease-modifying treatments. Transcription of the expanded allele produces toxic gain-of-function CUG-expansion RNA that sequesters the MBNL family of alternative splicing regulators into ribonuclear foci, leading to pathogenic mis-splicing. In this study, using our previously established HeLa DM1 repeat selective screening platform, we identified the natural flavonoid quercetin as a selective modulator of toxic CUG RNA levels. Quercetin treatment selectively reduced DMPK levels and rescued MBNL-dependent mis-splicing in DM1 patient-derived myotubes. To overcome the limited bioavailability of quercetin in vivo, we evaluated a bioavailable derivative of quercetin, enzymatically modified isoquercitrin (EMIQ), in the HSA LR DM1 skeletal muscle mouse model. EMIQ treatment of these mice through the drinking water selectively reduced the expanded CUG transcripts and rescued mis-splicing and myotonia. Given its efficacy in the HSA LR mouse model and the established safety profile in humans, we have identified EMIQ as a priority disease-targeting therapeutic lead for future clinical evaluation in DM1.

Individuals with cognitive disabilities have challenges with personal navigation and wayfinding, especially when traveling on public transportation. The purpose of this case study is to describe the structure and implementation of the Personal Navigation for Individuals with Disabilities (PNID) education and training program, which is based on a socio-technical architecture for individuals with cognitive disabilities within a fixed-route public bus system. A case study methodology was used to describe preliminary findings of the skills, attributes, and experiences of three individuals with cognitive disabilities as it relates to transportation on fixed-route bus systems in a midsized urban setting. The three individuals completed five training activities: safety, public bus, smartphone, WayFinder App, and fixed-route bus system. The case study provided a preliminary mixed-methods overview of training travelers with cognitive disabilities to use the WayFinder system while accessing fixed-route public bus system. The insights and strategies identified through the case study demonstrate the potential opportunities for development, implementation, and sustainability of the PNID program in other midsized urban settings. The PNID program (i.e. AT service delivery process), in combination with the WayFinder system (i.e. assistive technology), has the potential to meet the unique needs of individuals with cognitive disabilities when accessing public transportation.

Myotonic dystrophy type 1 (DM1) is considered a progeroid disease (i.e., causing premature aging). This hypervariable disease affects multiple systems, such as the musculoskeletal, central nervous, gastrointestinal, and others. Despite advances in understanding the underlying pathogenic mechanism of DM1, numerous gaps persist in our understanding, hindering elucidation of the heterogeneity and severity of its symptoms. Accumulating evidence indicates that the toxic intracellular RNA accumulation associated with DM1 triggers cellular senescence. These cells are in a state of irreversible cell cycle arrest and secrete a cocktail of cytokines, referred to as a senescence-associated secretory phenotype (SASP), that can have harmful effects on neighboring cells and more broadly. We hypothesize that cellular senescence contributes to the pathophysiology of DM1, and clearance of senescent cells is a promising therapeutic approach for DM1. We will discuss the therapeutic potential of different senotherapeutic drugs, especially senolytics that eliminate senescent cells, and senomorphics that reduce SASP expression.

CRISPR interference (CRISPRi) technologies have revolutionized bioengineering by providing precise tools for gene expression modulation, enabling targeted gene perturbation and metabolic pathway optimization. Despite these advances, achieving dynamic control over gene expression by CRISPR-based regulation remains a challenge due to its inherently static nature. Utilizing toehold-mediated strand displacement and ligand-responsive ribozymes (aptazymes), this study introduces switchable guide RNAs (gRNAs) that facilitate tunable gene expression mediated by mRNA or small molecule signals. We demonstrate complete silencing of gRNA via strategically designed 5' or 3' extensions that impede the gRNA spacer or the dCas9 handle, with subsequent restoration of function through sequestration or cleavage of the obstructive sequence. The resulting toehold-embedded or aptazyme-embedded gRNAs can be deactivated by specific signals, including two full-length translatable mRNAs and two small molecule triggers, thereby lifting CRISPRi repression on targeted genes. This modular approach allows for gRNA-based biocomputing through multi-layer or multi-input genetic logic gates in Saccharomyces cerevisiae. Offering a versatile strategy for post-CRISPR regulation in response to environmental signals or cellular states, this methodology expands the toolkit in eukaryotic systems for reversible control of gene expression.

There is increasing recognition of the need for patient-centred lupus self-management (SM) programmes to improve outcomes. The purpose of this manuscript is to describe the underlying methodology and initial pilot results of Strategies to Embrace Living with Lupus Fearlessly (SELF): an evidence-based digital SM intervention designed for individuals with lupus based on the Transtheoretical Model of Behaviour Change. We describe pilot testing and initial feasibility and acceptability results of SELF among individuals with lupus. Participants for the 90-day pilot test were recruited through three Centers for Disease Control & Prevention-funded lupus registry sites across the USA and the Lupus Foundation of America's constituency, including a diverse sociodemographic lupus population, using varied sources. Feasibility, acceptability and preliminary impact were assessed using data on enrolment, retention, ease of utilisation and satisfaction with SELF's features, changes in readiness for SM behaviours, impact on patient-reported outcomes (eg, fatigue interference) and user feedback. Nearly 80% of participants had moderate or high skill gaps at programme intake (ie, were in an early stage of change for a key SM skill), underscoring the importance of digital SM programmes like SELF. Among those who completed the programme, almost 60% of participants closed a skill gap by mastering one or more SM skills. Significant effects on patient-provider interactions and fatigue interference were also noted, highlighting a need for future investigation. Qualitative data were mostly positive in terms of feasibility and acceptability, with specific recommendations for future improvement. SELF utilisation and pilot data indicate that SELF is generally feasible and acceptable, particularly among those needing to build lupus SM skills. Future work will explore ways to improve the digital SM intervention and reduce barriers to initial and ongoing engagement.

Myotonic dystrophy type 1 (DM1), the leading cause of adult-onset muscular dystrophy, is caused by a CTG repeat expansion. Expression of the repeat causes widespread alternative splicing (AS) defects and downstream pathogenesis, including significant skeletal muscle impacts. The HSA LR mouse model plays a significant role in therapeutic development. This mouse model features a transgene composed of approximately 220 interrupted CTG repeats, which results in skeletal muscle pathology that mirrors DM1. To better understand this model and the growing number of therapeutic approaches developed with it, we performed a meta-analysis of publicly available RNA sequencing data for AS changes across three widely examined skeletal muscles: quadriceps, gastrocnemius, and tibialis anterior. Our analysis demonstrated that transgene expression correlated with the extent of splicing dysregulation across these muscles from gastrocnemius (highest), quadriceps (medium), to tibialis anterior (lowest). We identified 95 splicing events consistently dysregulated across all examined datasets. Comparison of splicing rescue across seven therapeutic approaches showed a range of rescue across the 95 splicing events from the three muscle groups. This analysis contributes to our understanding of the HSA LR model and the growing number of therapeutic approaches currently in preclinical development for DM1.

Maps have been invaluable navigation aids for millennia and thus have been critical for human survival. The increasing popularity of and high dependence on digital, location-aware assistive navigation technology, however, has been shown to divert our attention from the environment and to negatively influence innate spatial abilities. To mitigate this, neuroadaptive mobile geographic information displays (namGIDs) are proposed that respond in real-time to navigators' cognitive task demands and wayfinder's situated visuo-spatial attention needs. In doing so, namGIDs may not only help navigators maintain navigation efficiency but more importantly, also continuously scaffold spatial learning. To do this, the proposed navigation assistance must strike the appropriate balance between welcomed mobility efficiency gains while limiting human spatial deskilling. Leveraging neuroadaptive cartography, we can ensure to remain effective navigators, empowered to explore the world with confidence.

We describe the characteristics, content, and effectiveness of digital self-management (SM) education programs for lupus and other chronic conditions to identify gaps and inform the improvement of future programs in lupus. Three bibliographic databases were searched for articles published between May 2012 and April 2022. The search was cast to capture the breadth of digital SM education programs in the following conditions: lupus, epilepsy, fibromyalgia, multiple sclerosis, sickle cell anemia, Sjögren syndrome, psoriatic arthritis, and rheumatoid arthritis. Title and abstract screening, as well as full-text review, was conducted by two independent reviewers. Data extraction was first completed by one author charting all studies and then, a second time, by four members of the research team charting collaboratively. Of the 1,969 articles identified through the search, 14 met inclusion criteria. Two additional articles were included following bibliography review. The 16 articles represented 12 unique digital SM education programs. Programs covered five conditions: epilepsy (n = 3), fibromyalgia (n = 2), multiple sclerosis (n = 4), lupus (n = 1), and rheumatoid arthritis (n = 2). Most programs were asynchronous and internet-based (n = 9) with a prescribed sequence of content (n = 8). Peer, technical, or specialist support was offered in seven programs. Most programs demonstrated statistically significant improvement of symptoms in the intervention group (n = 8). This scoping review summarizes the current landscape for digital SM education programs in lupus and similar conditions. In lupus, further investigation will fill in the gaps around digital SM education needs, user experience, and evaluation of outcomes.

Engineering metabolism to efficiently produce chemicals from multi-step pathways requires optimizing multi-gene expression programs to achieve enzyme balance. CRISPR-Cas transcriptional control systems are emerging as important tools for programming multi-gene expression, but poor predictability of guide RNA folding can disrupt expression control. Here, we correlate efficacy of modified guide RNAs (scRNAs) for CRISPR activation (CRISPRa) in E. coli with a computational kinetic parameter describing scRNA folding rate into the active structure (rS = 0.8). This parameter also enables forward design of scRNAs, allowing us to design a system of three synthetic CRISPRa promoters that can orthogonally activate (>35-fold) expression of chosen outputs. Through combinatorial activation tuning, we profile a three-dimensional design space expressing two different biosynthetic pathways, demonstrating variable production of pteridine and human milk oligosaccharide products. This RNA design approach aids combinatorial optimization of metabolic pathways and may accelerate routine design of effective multi-gene regulation programs in bacterial hosts.

DNA nanostructures are typically assembled by thermal annealing in buffers containing magnesium. We demonstrate the assembly of DNA nanostructures at constant temperatures ranging from 4 °C to 50 °C in solutions containing different metal ions. The choice of metal ions and the assembly temperature influence the isothermal assembly of several DNA motifs and designed three-dimensional DNA crystals. Molecular dynamics simulations show more fluctuations of the DNA structure in select monovalent ions (Na+ and K+) compared to divalent ions (Mg2+ and Ca2+). A key highlight is the successful assembly of DNA motifs in nickel-containing buffer at temperatures below 40 °C, otherwise unachievable at higher temperatures, or using an annealing protocol. DNA nanostructures isothermally assembled in different ions do not affect the viability of fibroblasts, myoblasts, and myotubes and or the immune response in myoblasts. The use of ions other than the typically-used magnesium holds key potential in biological and materials science applications that require minimal amounts of magnesium.

Dynamic, multi-input gene regulatory networks (GRNs) are ubiquitous in nature. Multilayer CRISPR-based genetic circuits hold great promise for building GRNs akin to those found in naturally occurring biological systems. We develop an approach for creating high-performing activatable promoters that can be assembled into deep, wide, and multi-input CRISPR-activation and -interference (CRISPRa/i) GRNs. By integrating sequence-based design and in vivo screening, we engineer activatable promoters that achieve up to 1,000-fold dynamic range in an Escherichia coli-based cell-free system. These components enable CRISPRa GRNs that are six layers deep and four branches wide. We show the generalizability of the promoter engineering workflow by improving the dynamic range of the light-dependent EL222 optogenetic system from 6-fold to 34-fold. Additionally, high dynamic range promoters enable CRISPRa systems mediated by small molecules and protein-protein interactions. We apply these tools to build input-responsive CRISPRa/i GRNs, including feedback loops, logic gates, multilayer cascades, and dynamic pulse modulators. Our work provides a generalizable approach for the design of high dynamic range activatable promoters and enables classes of gene regulatory functions in cell-free systems.

Engineered living materials (ELMs) fabricated by encapsulating microbes in hydrogels have great potential as bioreactors for sustained bioproduction. While long-term metabolic activity has been demonstrated in these systems, the capacity and dynamics of gene expression over time is not well understood. Thus, we investigate the long-term gene expression dynamics in microbial ELMs constructed using different microbes and hydrogel matrices. Through direct gene expression measurements of engineered E. coli in F127-bisurethane methacrylate (F127-BUM) hydrogels, we show that inducible, input-responsive genetic programs in ELMs can be activated multiple times and maintained for multiple weeks. Interestingly, the encapsulated bacteria sustain inducible gene expression almost 10 times longer than free-floating, planktonic cells. These ELMs exhibit dynamic responsiveness to repeated induction cycles, with up to 97% of the initial gene expression capacity retained following a subsequent induction event. We demonstrate multi-week bioproduction cycling by implementing inducible CRISPR transcriptional activation (CRISPRa) programs that regulate the expression of enzymes in a pteridine biosynthesis pathway. ELMs fabricated from engineered S. cerevisiae in bovine serum albumin (BSA) - polyethylene glycol diacrylate (PEGDA) hydrogels were programmed to express two different proteins, each under the control of a different chemical inducer. We observed scheduled bioproduction switching between betaxanthin pigment molecules and proteinase A in S. cerevisiae ELMs over the course of 27 days under continuous cultivation. Overall, these results suggest that the capacity for long-term genetic expression may be a general property of microbial ELMs. This work establishes approaches for implementing dynamic, input-responsive genetic programs to tailor ELM functions for a wide range of advanced applications.