Objectives/Goals: Monensin is FDA approved for use in veterinary medicine. Recent studies pointed to its potent anticancer activity. Since de novo drug discovery process typically takes 10 to 15 years and requires an investment of approximately $1.3 to $3 billion, drug repositioning can bypass several steps in this process and increase the potential for success. Methods/Study Population: Cell viability assays were conducted on human MDA-MB-231, MDA-MB-468, and MCF10A breast cancer cell lines and mouse EO771 and 4T1 breast cancer cell lines. MDA-MB-231 cell line was used in all the studies unless specified otherwise. Time course levels of Bcl-2, Bak, p62, and LC3II were assessed via Western blotting with GAPDH as a loading control. Proteomics analysis was conducted by the IDEA National Resource for Quantitative Proteomics. Time course levels of major histocompatibility complex (MHC) I and II and calreticulin were evaluated using flow cytometry. At least three biological replicates have been conducted for each experiment. Results/Anticipated Results: Monensin and several of its novel analogs were potent toward human and mouse breast cancer cell lines. Furthermore, they induced apoptotic cell death as evidenced by Annexin V/PI assay, downregulation of Bcl-2, and upregulation of Bak in MDA-MB-231 cells. Proteomics analysis revealed that several molecular pathways related to MHC class I and II antigen presentation were significantly altered following treatment with these compounds. Additionally, monensin and its analogs significantly increased the expression of MHC class I and II. Our studies also showed that monensin and its analogs increase the surface calreticulin levels. Treatment of MDA-MB-231 cells with these compounds also resulted in an increase in p62 and LC3II expression, suggesting a disruption of the autophagic process. Discussion/Significance of Impact: These results suggest that monensin and its analogs not only exhibit anti-breast cancer cell activity but also modulate immune-related pathways. By disrupting autophagy and enhancing calreticulin levels, these compounds may potentiate antitumor immune responses, providing a promising avenue for drug repositioning in cancer therapy.

Objectives/Goals: Manual skin assessment in chronic graft-versus-host disease (cGVHD) can be time consuming and inconsistent (>20% affected area) even for experts. Building on previous work we explore methods to use unmarked photos to train artificial intelligence (AI) models, aiming to improve performance by expanding and diversifying the training data without additional burden on experts. Methods/Study Population: Common to many medical imaging projects, we have a small number of expert-marked patient photos (N = 36, n = 360), and many unmarked photos (N = 337, n = 25,842). Dark skin (Fitzpatrick type 4+) is underrepresented in both sets; 11% of patients in the marked set and 9% in the unmarked set. In addition, a set of 20 expert-marked photos from 20 patients were withheld from training to assess model performance, with 20% dark skin type. Our gold standard markings were manual contours around affected skin by a trained expert. Three AI training methods were tested. Our established baseline uses only the small number of marked photos (supervised method). The semi-supervised method uses a mix of marked and unmarked photos with human feedback. The self-supervised method uses only unmarked photos without any human feedback. Results/Anticipated Results: We evaluated performance by comparing predicted skin areas with expert markings. The error was given by the absolute difference between the percentage areas marked by the AI model and expert, where lower is better. Across all test patients, the median error was 19% (interquartile range 6 – 34) for the supervised method and 10% (5 – 23) for the semi-supervised method, which incorporated unmarked photos from 83 patients. On dark skin types, the median error was 36% (18 – 62) for supervised and 28% (14 – 52) for semi-supervised, compared to a median error on light skin of 18% (5 – 26) for supervised and 7% (4 – 17) for semi-supervised. Self-supervised, using all 337 unmarked patients, is expected to further improve performance and consistency due to increased data diversity. Full results will be presented at the meeting. Discussion/Significance of Impact: By automating skin assessment for cGVHD, AI could improve accuracy and consistency compared to manual methods. If translated to clinical use, this would ease clinical burden and scale to large patient cohorts. Future work will focus on ensuring equitable performance across all skin types, providing fair and accurate assessments for every patient.

Objectives/Goals: Osteoarthritis (OA) is a multifactorial disease where sustained gut inflammation is a continued source of inflammatory mediators driving degenerative processes in joints. The goal was to use spontaneous equine model to compare fecal and leukocyte microbiome and correlation to transcriptome in OA. Methods/Study Population: Seventy-six horses (31 OA, 45 controls) were enrolled by population-based sampling. Feces and peripheral blood mononuclear cells (PBMC) were collected. Horses were determined to have OA by clinical and radiographic evidence. Horses were excluded if they received medications or joint injections within two months. Fecal and circulating leukocyte bacterial microbial 16s-seq was performed. Bulk RNAseq of PBMC was performed by the Illumina platform. Gene expression data were mapped to the equine genome, and differential expression analysis was performed with DESeq2. Qiime2 was used for microbial analysis. Enrichment analysis was performed with a cluster profiler. Correlation analyses were performed between the datasets. Results/Anticipated Results: Beta and alpha microbial diversity differed in feces and PBMC of OA vs. healthy horses. Horses with OA had an increased Firmicutes to Bacteroidetes ratio compared with controls. The fecal microbiome of OA horses had significantly higher amounts of Firmicutes Oribacterium (q Discussion/Significance of Impact: These data suggest that altered microbiome and PBMC gene expression are associated with naturally occurring OA in the translational equine model. While Oribacterium has been detected in humans with rheumatoid arthritis, its role in OA warrants further proteomic and metabolomic profiling.

Objectives/Goals: Lung transplant is a life-saving surgery for patients with advanced lung diseases yet long-term survival remains poor. The clinical features and lung injury patterns of lung transplant recipients who die early versus those who survive longer term remain undefined. Here, we use cell-free DNA and rejection parameters to help elucidate this further. Methods/Study Population: Lung transplant candidacy prioritizes patients who have a high mortality risk within 2 years and will likely survive beyond 5 years. We stratified patients who died within 2 years of transplant as early death (n = 50) and those who survived past 5 years as long-term survivors (n = 53). Lung transplant recipients had serial blood collected as part of two prospective cohort studies. Cell-free DNA (cfDNA) was quantified using relative (% donor-derived cfDNA {%ddcfDNA}) and absolute (nuclear-derived {n-cfDNA}, mitochondrial-derived {mt-cfDNA}) measurements. As part of routine posttransplant clinical care, all patients underwent pulmonary function testing (PFT), surveillance bronchoscopy with bronchoalveolar lavage (BAL), transbronchial biopsy (TBBx), and donor-specific antibody testing (DSA). Results/Anticipated Results: Over the first 2 years after transplant, the number of episodes of antibody-mediated rejection (p) Discussion/Significance of Impact: Clinically, early-death patients perform worse on routine surveillance PFTs and experience a worse degree of CLAD. These patients also have higher levels of cfDNA as quantified by n-cfDNA and mt-cfDNA. These results provide preliminary evidence that early-death patients have worse allograft rejection, dysfunction, and molecular injury.

Since the 1950s, the United Nations (UN) has designated days (e.g., World Wetland Day), years (e.g., Year of the Gorilla) and decades (e.g., Decade on Biodiversity) with a commonly stated goal to raise awareness and funding for conservation-oriented initiatives, and these Days, Years and Decades of ‘…’ (hereafter ‘DYDOs’) continue. However, the effectiveness of these initiatives to achieve their stated objectives and to contribute to positive conservation outcomes is unclear. Here we used a binary analysis change model to evaluate the effectiveness of UN conservation-oriented DYDOs observed between 1974 and 2020. We also examined four case studies to understand the different strategies employed to meet specified conservation goals. We found that DYDOs apparently contributed to positive conservation outcomes when they were tied to social media campaigns and/or when they were strategically situated in current events or global discourse. Although the outcomes of DYDOs were varied, those with longer timescales and those that engaged local communities were more likely to be successful. We suggest that DYDO organizers should identify all possible paths of action through the lens of the change model outlined in this paper to strengthen the value and effectiveness of these initiatives in the future. Using this approach could help ensure that resources are used efficiently and effectively, and that initiatives yield positive conservation outcomes that benefit people and nature.

Maladaptive daydreaming is a distinct syndrome in which the main symptom is excessive vivid fantasising that causes clinically significant distress and functional impairment in academic, vocational and social domains. Unlike normal daydreaming, maladaptive daydreaming is persistent, compulsive and detrimental to one’s life. It involves detachment from reality in favour of intense emotional engagement with alternative realities and often includes specific features such as psychomotor stereotypies (e.g. pacing in circles, jumping or shaking one’s hands), mouthing dialogues, facial gestures or enacting fantasy events. Comorbidity is common, but existing disorders do not account for the phenomenology of the symptoms. Whereas non-specific therapy is ineffective, targeted treatment seems promising. Thus, we propose that maladaptive daydreaming be considered a formal syndrome in psychiatric taxonomies, positioned within the dissociative disorders category. Maladaptive daydreaming satisfactorily meets criteria for conceptualisation as a psychiatric syndrome, including reliable discrimination from other disorders and solid interrater agreement. It involves significant dissociative aspects, such as disconnection from perception, behaviour and sense of self, and has some commonalities with but is not subsumed under existing dissociative disorders. Formal recognition of maladaptive daydreaming as a dissociative disorder will encourage awareness of a growing problem and spur theoretical, research and clinical developments.

Beliefs about other players’ strategies are crucial in determining outcomes for coordination games. If players are to coordinate on an efficient equilibrium, they must believe that others will coordinate with them. In many settings there is uncertainty about beliefs as well as strategies. Do people consider these “higher-order” beliefs (beliefs about beliefs) when making coordination decisions? I design a modified stag hunt experiment that allows me to identify how these higher-order beliefs and uncertainty about higher-order beliefs matter for coordination. Players prefer to invest especially when they believe that others are “optimistic” that they will invest; but knowledge that others think them unlikely to invest does not cause players to behave differently than when they do not know what their partners think about them. Thus resolving uncertainty about beliefs can result in marked efficiency gains.

We replicate an influential study of monetary incentive effects by Jamal and Sunder (1991) to illustrate the difficulties of drawing causal inferences from a treatment manipulation when other features of the experimental design vary simultaneously. We first show that the Jamal and Sunder (1991) conclusions hinge on one of their laboratory market sessions, conducted only within their fixed-pay condition, that is characterized by a thin market and asymmetric supply and demand curves. When we replicate this structure multiple times under both fixed pay and pay tied to performance, our findings do not support Jamal and Sunder's (1991) conclusion about the incremental effects of performance-based compensation, suggesting that other features varied in that study likely account for their observed difference. Our ceteris paribus replication leaves us unable to offer any generalized conclusions about the effects of monetary incentives in other market structures, but the broader point is to illustrate that experimental designs that attempt to generalize effects by varying multiple features simultaneously can jeopardize the ability to draw causal inferences about the primary treatment manipulation.

We present a 1000 km transect of phase-sensitive radar measurements of ice thickness, basal reflection strength, basal melting and ice-column deformation across the Ross Ice Shelf (RIS). Measurements were gathered at varying intervals in austral summer between 2015 and 2020, connecting the grounding line with the distant ice shelf front. We identified changing basal reflection strengths revealing a variety of basal conditions influenced by ice flow and by ice–ocean interaction at the ice base. Reflection strength is lower across the central RIS, while strong reflections in the near-front and near-grounding line regions correspond with higher basal melt rates, up to 0.47 ± 0.02 m a−1 in the north. Melting from atmospherically warmed surface water extends 150–170 km south of the RIS front. Melt rates up to 0.29 ± 0.03 m a−1 and 0.15 ± 0.03 m a−1 are observed near the grounding lines of the Whillans and Kamb Ice Stream, respectively. Although troublesome to compare directly, our surface-based observations generally agree with the basal melt pattern provided by satellite-based methods but provide a distinctly smoother pattern. Our work delivers a precise measurement of basal melt rates across the RIS, a rare insight that also provides an early 21st-century baseline.

People with serious mental illness (SMI) are over-represented throughout the US criminal justice system. To address this issue, forensic assertive community treatment has recently emerged as a best-practice intervention. Also known as forensic ACT, ForACT, or most commonly as “FACT,” forensic assertive community treatment is an adaptation of the assertive community treatment (ACT) model. Unlike ACT, however, FACT is purposefully designed to prevent arrest and incarceration among people with SMI who have histories of involvement with the criminal justice system (i.e., “justice-involved” individuals). Although FACT was recognized as a best practice by the Substance Abuse and Mental Health Services Administration (SAMHSA) in 2019, FACT teams vary widely in their structure and daily operations. This lack of a standard FACT model continues to impede FACT program implementation and outcomes research. This article begins with a review of FACT origins, followed by a discussion of what we know (and do not know) about FACT operation and effectiveness. Based on the authors’ experience, the article then discusses key components of FACT and concludes with a discussion of current challenges and research recommendations for FACT model development.

Stauffer (2021) shows that when Americans perceive their legislatures as having more gender diversity, they perceive these institutions to be more responsive. In this letter, we use an independently fielded survey to validate Stauffer’s findings and investigate whether her findings extend to race. We successfully replicate Stauffer’s analyses. We also newly demonstrate that perceptions of Black representation in legislatures are related to increased external efficacy, and perceptions of gender descriptive representation are associated with increased trust. Such findings underscore the importance of studying citizens’ perceptions of descriptive representation to understand their attitudes toward government.

Digital twins are a new paradigm for our time, offering the possibility of interconnected virtual representations of the real world. The concept is very versatile and has been adopted by multiple communities of practice, policymakers, researchers, and innovators. A significant part of the digital twin paradigm is about interconnecting digital objects, many of which have previously not been combined. As a result, members of the newly forming digital twin community are often talking at cross-purposes, based on different starting points, assumptions, and cultural practices. These differences are due to the philosophical world-view adopted within specific communities. In this paper, we explore the philosophical context which underpins the digital twin concept. We offer the building blocks for a philosophical framework for digital twins, consisting of 21 principles that are intended to help facilitate their further development. Specifically, we argue that the philosophy of digital twins is fundamentally holistic and emergentist. We further argue that in order to enable emergent behaviors, digital twins should be designed to reconstruct the behavior of a physical twin by “dynamically assembling” multiple digital “components”. We also argue that digital twins naturally include aspects relating to the philosophy of artificial intelligence, including learning and exploitation of knowledge. We discuss the following four questions (i) What is the distinction between a model and a digital twin? (ii) What previously unseen results can we expect from a digital twin? (iii) How can emergent behaviours be predicted? (iv) How can we assess the existence and uniqueness of digital twin outputs?

We report experiments investigating how experience influences the endowment effect. Our experiments feature endowments which are bundles of unfamiliar consumption goods. We examine how a subject’s willingness to swap items from their endowment is influenced by prior experiences of tasting the goods in question and by prior experiences of choosing between them. We do not find a statistically significant endowment effect in our baseline treatment and, because of this, we are unable to test for an effect of consumption experience. We do find an endowment effect when the endowment is acquired in two instalments and, in this setting, we find some evidence that choice experience increases trading. In a follow-up experiment, we find evidence that the absence of an endowment effect in our baseline treatment is due to subjects being more willing to swap when they do not have to give up the last unit of their endowment.