OBJECTIVES/GOALS: During and after cancer treatment, cancer-related fatigue (CRF) is a debilitating symptom reported by up to 80% of cancer patients Our understanding of the pathology underlying CRF is limited. Preliminary RNA sequencing data suggest that increased levels of KIR3DL1, the natural killer cell (NK) immunoglobulin-like receptor 3DL1a, may be associated with CRF. METHODS/STUDY POPULATION: Fatigue was measured using the Functional Assessment of Chronic Illness Therapy-Fatigue (FACT-F). Functional validation of the NK cell finding was performed from whole blood obtained from fatigued and non-fatigued subjects. NK cells were isolated from freshly collected whole blood using a human NK cell isolation kit based on CD56 microbead positive selection. NK cell function was assessed using the NK cell direct cytotoxicity assay. Briefly, isolated NK cells were co-cultured in a 2:1 ratio with calcein AM-labelled K562 cells, which are NK cell-sensitive due to the very low MHCI expression. NK cell-mediated cytotoxicity was assessed with Cytation 1 Cell Imaging Multi-Mode reader. Flow cytometric protocols were used to examine NK subset differences between the fatigued and non-fatigued groups. RESULTS/ANTICIPATED RESULTS: NK cells isolated from the fatigued group exhibited decreased cytotoxicity at 12.28% compared to NK cells isolated from non-fatigued controls at a mean of 40.6% cytotoxicity. Flow cytometry analysis revealed a decrease in the CD56dim CD16bright population in the fatigued group (87.1% of CD56+CD4- cells) compared to the control (91.4% of CD56+CD4- cells). Furthermore, there was a decrease in NKG2A expression in mature NK cells (CD56dim CD16bright) isolated from the fatigued group compared to the non-fatigued group. DISCUSSION/SIGNIFICANCE OF IMPACT: Results from the pilot study suggest that there was a decrease in NK cell cytotoxicity in the fatigued group. In addition, there may be a shift in NK cell subpopulations associated with fatigue. Findings from this pilot study suggest that impaired NK cell function may be associated with CRF pathogenesis.

ABSTRACT IMPACT: Brain networks can be explored by delivering brief pulses of electrical current in one area while measuring responses in other areas, and this describes an open-source novel algorithm to carry out this exploration. OBJECTIVES/GOALS: If we focus on a single brain site and observe the average effect of stimulating each of many other brain sites, visually-apparent motifs in the temporal response shape emerge from adjacent stimulation sites. There are no existing approaches to identify and quantify the spatiotemporal structure of these motifs. METHODS/STUDY POPULATION: Individual stimulation trials are correlated with one another, then a correlation-significance matrix quantifying similarity between stimulation sites is decomposed with non-negative matrix factorization, in which the inner dimension is iteratively reduced. The dimensionality reduction identifies stimulation sites that produce a common elicited temporal response, and linear kernel PCA is applied to obtain the robust profile of this response cluster. RESULTS/ANTICIPATED RESULTS: We describe and illustrate a data-driven approach to determine characteristic spatiotemporal structure in these response shapes, summarized by a set of unique ‘basis profile curves’ (BPCs). Each BPC may be mapped back to underlying anatomy in a natural way, quantifying projection strength from each stimulation site using simple metrics. Our technique is demonstrated for an array of implanted brain surface electrodes in a human patient, and our code is shared at https://purl.stanford.edu/rc201dv0636. DISCUSSION/SIGNIFICANCE OF FINDINGS: This framework enables straightforward interpretation of single-pulse brain stimulation data, and can be applied generically to explore the diverse milieu of interactions that comprise the connectome.

OBJECTIVES/GOALS: We aimed to find the histology-specific markers that were predictive of post-operative long-term chordoma recurrence (≤1 year) using trained multiple tree-based machine learning (ML) algorithms. METHODS/STUDY POPULATION: We reviewed the records of patients who had treatment for clival and spinal chordomas between January 2017 and June 2021 across the Mayo Clinic enterprise (Minnesota, Florida, and Arizona). Patients were excluded if they had no histopathology or recurrence as an outcome. Demographics, type of treatment, clinical and radiological follow-up duration, histopathology, and other relevant clinical factors were abstracted from each patient record. Decision tree and random forest classifiers were trained and tested to predict the long-term recurrence based on unseen data using an 80/20 split. The performance of the model was optimized using tenfold cross-validation. RESULTS/ANTICIPATED RESULTS: One hundred fifty-one patients were identified: 58 chordomas from the clivus, 48 chordomas of the mobile spine, and 45 sacrococcygeal. Subtotal Resection followed by radiation therapy, was the most common treatment modality, followed by Gross Total resection, then radiation therapy. The multivariate analysis defines the molecular predictors of recurrence following resection. S100 and pan-cytokeratin is more likely to increase the risk of post-operative recurrence (OR= 3.67; CI= [1.09,12.42], p=0.03). In the decision tree analysis, a clinical follow-up > 1897 days was found in 37 % of encounters and a 90% chance of being classified for recurrence (Accuracy= 77%). Factors predicting long-term recurrence are the patient’s age, type of surgical treatment, location of the tumor, S100, pan-cytokeratin, and EMA. DISCUSSION/SIGNIFICANCE: Our molecular and clinicopathological variables combined with tree-based ML tools successfully demonstrate a high capacity to individually identify the patient’s recurrence pattern with an accuracy of 77%. S100, pan-cytokeratin, and EMA were the histologic drivers of recurrence.

Objectives/Goals: Magnetic resonance imaging (MRI) reports are stored as unstructured text in the electronic health record (EHR), rendering the data inaccessible. Large language models (LLM) are a new tool for analyzing and generating unstructured text. We aimed to evaluate how well an LLM extracts data from MRI reports compared to manually abstracted data. Methods/Study Population: The University of California, San Francisco has deployed a HIPAA-compliant internal LLM tool utilizing GPT-4 technology and approved for PHI use. We developed a detailed prompt instructing the LLM to extract data elements from prostate MRI reports and to output the results in a structured, computer-readable format. A data pipeline was built using the OpenAI Application Programming Interface (API) to automatically extract distinct data elements from the MRI report that are important in prostate cancer care. Each prompt was executed five times and data were compared with the modal responses to determine variability of responses. Accuracy was also assessed. Results/Anticipated Results: Across 424 prostate MRI reports, GPT-4 response accuracy was consistently above 95% for most parameters. Individual field accuracies were 98.3% (96.3–99.3%) for PSA density, 97.4% (95.4–98.7%) for extracapsular extension, 98.1% (96.3–99.2%) for TNM Stage, had an overall median of 98.1% (96.3–99.2%), a mean of 97.2% (95.2–98.3%), and a range of 99.8% (98.7–100.0%) to 87.7% (84.2–90.7%). Response variability over five repeated runs ranged from 0.14% to 3.61%, differed based on the data element extracted (p Discussion/Significance of Impact: GPT-4 was highly accurate in extracting data points from prostate cancer MRI reports with low upfront programming requirements. This represents an effective tool to expedite medical data extraction for clinical and research use cases.

Objectives/Goals: Pulmonary infections by Staphylococcus aureus (S. aureus) are difficult to eradicate in people with cystic fibrosis (CF, PwCF) and remain a significant disease burden. Our objective is to determine if a MEK1/2 inhibitor, ATR-002, can reduce S. aureus biofilm growth and synergize with antibiotics against multidrug-resistant strains of S. aureus. Methods/Study Population: To quantify the ability of ATR-002 to synergize with antibiotics, synergy checkerboard assays examined the effects of ATR-002 in combination with n=11 different antibiotics against the methicillin-resistant S. aureus (MRSA) strain USA300. We next quantified bacterial growth of n=6 antibiotic-resistant CF clinical isolates treated with a combinatory dose of 5 uM ATR-002 and antibiotic to confirm results from the checkerboard assay. Finally, S. aureus biofilms were established using USA300 or n=7 different CF clinical isolates in 96-well plates for 24 hours and then were treated with media, vehicle controls, or ATR-002 (5, 25, or 50 uM) for an additional 24 hours; bacterial biomass was quantified by crystal violet staining to assess the ability of ATR-002 to decrease biofilm growth. Results/Anticipated Results: Synergy checkerboard assays revealed that ATR-002 can synergize with the antibiotics gentamicin and amikacin, but had additive/indifferent effects with clindamycin, erythromycin, nafcillin, vancomycin, doxycycline, daptomycin, sulfamethoxazole-trimethoprim, linezolid, and puromycin. Additionally, two gentamicin/amikacin-resistant CF isolates were resensitized to gentamicin and amikacin with the addition of 5 uM ATR-002. However, CF isolates resistant to erythromycin, clindamycin, and nafcillin could not be resensitized to antibiotics with the addition of 5 uM ATR-002. Biofilms produced by MRSA, and n=3 out of 7 CF clinical isolates tested were significantly reduced by treatment of 50 uM ATR-002. Discussion/Significance of Impact: Our results demonstrate that ATR-002 can synergize with some antibiotics against MRSA, which is also observed in antibiotic-resistant CF S. aureus clinical isolates, and that ATR-002 can reduce established S. aureus biofilms. Future studies will explore the antibacterial effects of ATR-002 in additional clinically relevant models of infection.

OBJECTIVES/SPECIFIC AIMS: Engaging Richmond is a community-university partnership, made up of local residents and university faculty and staff that was established in 2011 with an NIH supplement to a Clinical and Translational Science Research Award at Virginia Commonwealth University (VCU). The primary aims of the supplement were to (1) to conduct community-based participatory research (CBPR) on the leading causes of health disparities perceived by the Richmond community and (2) to thereby highlight community needs and assets and build capacity for future community-engaged research (CEnR). The goal was to prepare a community-focused, community-prioritized, health equity report while building capacity, strengthening relationships, and discovering local barriers to CEnR, and therefore to stimulate, facilitate, and inform future CEnR at VCU. METHODS/STUDY POPULATION: This is a case study exploring the impact of 1 community-university partnership on investigator-initiated research using historical and qualitative data. RESULTS/ANTICIPATED RESULTS: Although Engaging Richmond received only 12 months of support from the NIH supplement that provided its initial funding, the community-university partnership has worked continuously since its formation in 2011. This work has not only helped to build connections with the community and key stakeholders, it has also contributed substantially to the resources available to university faculty pursuing CEnR. Specifically, we find that Engaging Richmond has contributed to investigator initiated research in the following ways, either working as co-investigators or in a consultative capacity: consultation on proposal development (5 projects); assisted with instrument development (4 projects); participant recruitment (7 projects); data collection and analysis (6 projects); dissemination (5 projects). In addition to collaboration on projects, Engaging Richmond has increased institutional capacity for CEnR through its contributions to the Annual Community Engaged Institute at the university and the Center of Clinical and Translational Science’s Community Review Board (CRB). The CRB helps researchers work successfully in a community setting, enhance the research design, help to improve study implementation and assist with translation and dissemination of findings. DISCUSSION/SIGNIFICANCE OF IMPACT: Although community-university partnerships have become much more common over the past several decades, there remains a gap in research evidence on the impact of these partnerships. In their 2004 review, Viswanathan et al. note that community-based participatory research studies infrequently document improved capacity of researchers and research organizations as an outcome, despite the expectation that such improvement will accrue through investment in CEnR. A more recent study assessing the range of community-university partnerships across a research university also noted the lack of processes in place to assess impacts (Holton et al., 2015). While assessments of CEnR impact on communities have become increasingly common as demand for evidence about the effectiveness of community-engaged partnerships has mounted, there does not appear to be a similar trend in assessing the impact of these efforts on faculty research and institutional capacity. By focusing on the impact of 1 community-university partnership that has been sustained for over 5 years, we highlight the ways in which having ongoing partnerships in place can support and strengthen investigator-initiated research, reflecting the flexible, “2-way approach” (Weerts and Sandmann, 2010) at the heart of CEnR.

ABSTRACT IMPACT: Pharmacological activation of KATP channels may provide analgesia and attenuate opioid tolerance and withdrawal OBJECTIVES/GOALS: Our long term goal is to develop therapeutics for the treatment of the overuse of opioids. The objective of this application is to test novel KATP channel-targeting prodrugs in rodent models of neuropathic and inflammatory pain in addition to opioid tolerance after chronic morphine administration. METHODS/STUDY POPULATION: In one study, two different measures for chronic pain were implemented in mice. Male and female mice (n=10) were subjected to spinal nerve ligation (SNL) or intraplantar injection of Complete Freund’s Adjuvant (CFA) to induce neuropathic and inflammatory pain, respectively. Administration of KATP channel prodrugs (60ug, it) attenuated mechanical hypersensitivity after SNL or CFA compared to vehicle (saline). In a separate study, changes in mechanical hypersensitivity were tested while mice undergo chronic morphine treatment (15mg/kg, 2x, 5 days) with administration of the prodrugs. Tolerance was measured as the loss of antinociception, and withdrawal is measured ˜24 hours after the final morphine injection. RESULTS/ANTICIPATED RESULTS: Intrathecal administration of either KATP channel prodrugs significantly attenuated mechanical hypersensitivity after SNL and significantly attenuated mechanical hypersensitivity after CFA in mice. We predict that intrathecal administration of these prodrugs will also attenuate morphine tolerance and withdrawal in mice. This hypothesis is based off our previous data indicating non-water soluble KATP channel agonists produce analgesia and attenuate morphine tolerance in mice. DISCUSSION/SIGNIFICANCE OF FINDINGS: Pharmaceutical strategies to utilize KATP channels for therapeutics have been hindered due to the low solubility and low ability to cross the neurovascular unit. Newly developed, water-soluble KATP channel openers could be useful pharmaceutical strategy to reduce chronic pain, opioid tolerance, and withdrawal in human populations.

OBJECTIVES/GOALS: 1. Describe the importance of community engaged research. 2. Explore the impact of participating in Community Engagement Studios from the perspective of the community expert. 3. Explain how Community Engagement Studios can positively impact clinical and translational sciences. METHODS/STUDY POPULATION: Stakeholders who participate in CE Studios are defined as community experts (CEs); care-givers, patients, researchers, etc. They are not classified as research participants, but rather consultants, who are designated with the task of providing input on the specified research study. CEs were recruited in a variety of formats such as clinics, social media, and other community-based settings. To be eligible for the study, CEs had to participate in at least one CE Studio prior to data collection. Data was collected from an online survey from April to September of 2019. One hundred and fifteen CEs participated in the study. We conducted thematic analysis on a total of 233 participant responses to three open-ended questions assessing attitudes towards research, desire to engage in research, and understanding of research. RESULTS/ANTICIPATED RESULTS: Ten major themes emerged from the data: involvement, togetherness, trust, value, confidence, community engagement, community connectedness, encouraging others to participate, increase knowledge and awareness, and respect. A major overarching theme of inclusion was presented in the data. Findings from this study indicate that CE Studios provide a space for CEs to gain a better understanding of the multi-faceted research process, offer insight into ways to include historically excluded populations, and increase trust, confidence and respect for researchers and the research process. Moreover, community experts felt connected to their community by participating in CE Studios and expressed interest in encouraging others to participate. DISCUSSION/SIGNIFICANCE: CEs are a crucial component of the research process, and their experiences should be utilized and reflected upon as we work to reduce health disparities. CE Studios serve to build trust, identify strategic areas of improvement in research, and provide a foundation to introduce community members to the multiple ways to engage in research.

OBJECTIVES/SPECIFIC AIMS: The Clinical and Translational Science Award (CTSA) program is a national consortium of 50+ academic medical research centers charged with accelerating the translation of clinical research. In 2017, the NIH National Center for Advancing Translational Sciences anticipates total CTSA program funding of over $500M. The consortium’s hub-and-spoke structure makes it a natural dissemination network, and the newest funding announcement makes dissemination of innovation across the consortium an explicit goal, but characteristics of CTSA hubs as adopters and transmitters of innovation are unknown. METHODS/STUDY POPULATION: A content analysis was conducted using data from CTSA hub Web sites (n=64) and a structured coding taxonomy based on 6 constructs drawn from literature about diffusion of innovation in service organizations (Greenhalgh et al., 2004): dissemination priority, institutional complexity, communication infrastructure, support for dissemination/implementation functions, cross-institutional collaboration/networking, and leadership composition. RESULTS/ANTICIPATED RESULTS: In total, 52% of hubs will renew under the new PAR in the next few years, providing an incentive to demonstrate dissemination capacity (although hubs will likely lag in operationalizing these activities until they are funded). A third of hubs (34%) represent more than one academic/research institution, and almost 80% of hubs have more than one clinical affiliate. To accommodate these different levels of institutional complexity, broad diffusion will require multi-modal, locally adapted dissemination efforts. Only 25% of hubs have capacity to undertake additional dissemination activities, and only 27% provide formal D&I support, suggesting that additional capacity/support will be needed to operationalize the CTSA dissemination mission. In total, 30% of hubs participate in cross-institutional collaboration/networking, so many may not have existing norms/tools supporting inter-institutional collaboration, but 77% include leadership from outside the School of Medicine, facilitating effective intrainstitutional dissemination. DISCUSSION/SIGNIFICANCE OF IMPACT: Understanding more about CTSA hubs as both adopters and transmitters of innovation can facilitate strategic use of these sites as a built-in dissemination network to amplify the reach and impact of clinical innovation and improve population health. Based on this initial analysis, the CTSA network does not appear to be fully primed for broad, rapid dissemination of innovation across its sites. In-depth interviews are being conducted to investigate CTSA hubs’ perceptions of their dissemination capacity and roles as adopters and transmitters of innovation.

OBJECTIVES/SPECIFIC AIMS: The objectives of this study are to determine whether high-frequency ipsi-lesion or low-frequency contra-lesion ECS improves forelimb function following experimental stroke in aged animals with focal and large strokes. We also want to investigate whether ECS-induced improvements in motor function are related to an enhancement of neural structural plasticity (dendrites and synapses) and changes in growth promoting (BDNF) and growth inhibiting (NOGO-A) expression in the infarcted motor cortex in young and aged animals. METHODS/STUDY POPULATION: We will investigate whether excitatory ECS of the infarcted cortex or inhibition of the noninfarcted cortex combined with daily impaired-forelimb rehabilitative training (RT) results in greater motor functional recovery compared to RT alone. Immunohistochemical (IHC) analyses and unbiased stereological techniques will be performed to investigate changes in proteins associated with dendritic restructuring (MAP2), synaptic plasticity (PSD95 and synaptophysin), and alteration in the expression of BDNF and NOGO-A. RESULTS/ANTICIPATED RESULTS: We expect that inhibitory ECS of the noninfarcted motor cortex will improve behavioral outcomes in moderate to severe stroke animals compared with excitatory ECS or no stimulation (RT alone) animals. We predict that the ECS condition that improves motor performance most significantly compared with RT alone will have a corresponding greater increase in remaining ipsi-infarct motor cortical dendritic and synaptic plasticity (demonstrated by a greater density of MAP2, synaptophysin, and PSD-95 immunoreactivity), and greater expression of BDNF. It is unknown, but also expected that better behavioral recovery will coincide with a greater reduction in NOGO-A in the injured motor cortex. DISCUSSION/SIGNIFICANCE OF IMPACT: These studies will aid in creating a model that will allow for a better understanding of the relationship between brain stimulation, severity of injury and, in future studies, aging. These studies will also help clarify previous conflicting brain stimulation results.

OBJECTIVES/SPECIFIC AIMS: The cellular mechanisms that underlie the IFNβ/α ratio that predicts response are not known. Effects of IFN on single immune cells may be masked in whole blood or mixed cell populations. By studying the effect of IFNβ/α activity ratio on individual monocytes, we can determine the functional impact of the IFN ratio and suggest the cellular mechanisms that underlie response/non-response to TNFi therapy in RA. METHODS/STUDY POPULATION: We used single cell analysis to investigate whether monocyte gene expression differs significantly between RA patients according to their pre-TNFi serum IFN-β/α ratio. Single classical (CL) and non-classical (NC) blood-derived monocytes were isolated from 15 seropositive RA subjects prior to biologic therapy. Subjects were grouped by pre-TNFi serum IFN-β/α ratio into two groups, those with a high IFN-β/α ratio (≥1.3, n = 6) and those with a low IFN-β/α ratio (<1.3, n = 9). 87 target genes were analyzed. Genes that varied significantly between the groups by categorical analyses were tested in multivariate logistic regression models. RESULTS/ANTICIPATED RESULTS: Every participant was seropositive for rheumatoid factor and antibodies to cyclic citrullinated peptide. Among the participants in the groups, there were no significant differences in age or DAS scores (P>0.05). The treatments were comparable and none were being treated with biologic therapy. There were striking differences in monocyte gene expression between patients with pre-treatment blood IFNβ/α activity <1.3 and ≥1.3. Expression of (1) key type I IFN pathway genes (JAK1, STAT2, IFIT2, IFIH1, PRDM1); (2) IL12; (3) CD36; and (4) CTLA4 were the strongest differentiators between groups (p<0.0001 for each, corrected for multiple comparisons). DISCUSSION/SIGNIFICANCE OF IMPACT: In this study we were able to measure gene expression in single monocytes from seropositive RA patients prior to biologic treatment. Within-cell co-expression patterns demonstrate biological differences in monocytes of RA patients with an IFNβ/α ≥1.3, the ratio of type I IFNs which predicts non-response to TNFi. The data suggest that there may be differential IFN production and pathway activation in patients who do not respond to TNFi. The increased expression of CD36 in monocytes from RA patients with high IFN β/α activity may be a reflection of increased “foam cells” in the inflamed tissue of patients who do not respond to TNFi. Enrichment of CTLA4 in those with high serum IFNβ/α suggests that CTLA4-Ig may be less likely to be an effective alternative for someone who is not likely to respond to TNFi. Current work includes determining whether the peripheral blood findings reflect altered cellular composition, type I IFN production and signaling in the synovium. Significance: This work will help to develop a more individualized approach to therapy in RA and determine an immunological basis of response/non-response to TNFi.

ABSTRACT IMPACT: We aim to understand how LTCCs impact cerebellar function. OBJECTIVES/GOALS: L-type calcium channels (LTCCs) are important in activity-dependent neurite outgrowth, which comprises neurite initiation and elongation. We used cerebellar granule neurons (CGNs) to differentiate between LTCC effects on neurite initiation vs elongation. We also tested cerebellar function in mice lacking specific LTCCs with behavioral assays. METHODS/STUDY POPULATION: CGNs were cultured from 129SvEv mouse pups at P4-P6. Potassum chloride (50mM) was used to stimulate neuronal cultures for 24 hours. Isradipine (20nM) was added to culture medium to inhibit all LTCCs for 1 hour. For Cav1.2 deletion, we crossed Cav1.2 conditional knockout mice (Cav1.2-cKO) to Syn-Cre mice (for deletion in most neurons) or Atoh1-Cre mice (for deletion in CGNs). The Cav1.2-cKO line was maintained on a 129SvEv background. For constitutive Cav1.3 deletion, mice were maintained on a C57BL/6NTac. Behavioral tasks included open field, rotarod, and Erasmus Ladder. Data were analyzed with sexes combined and separated to assess for sex as a biological variable. Studies were analyzed by one-way ANOVA, two-way ANOVA, or generalized linear mixed model, where appropriate. RESULTS/ANTICIPATED RESULTS: CGNs exhibited an increase in neurite initiation but not elongation when stimulated with potassium chloride, consistent with previous reports of activity-dependent neurite outgrowth in this cell type. LTCC inhibition with isradipine blunted KCl-induced neurite initiation. We observed no change in the length of either primary or secondary neurites with isradipine treatment with or without KCl stimulation. In our behavioral experiments, we observed no deficits in open field, rotarod, or Erasmus Ladder when Cav1.2 was deleted in most neurons (driven by Syn-Cre expression) or in cerebellar granule neurons (driven by Atoh1-Cre expression). In contrast, loss of Cav1.3 was associated with impaired motor learning in the rotarod task without evidence of ataxia on Erasmus Ladder. DISCUSSION/SIGNIFICANCE OF FINDINGS: We show a specific role for LTCCs in activity-dependent CGN neurite initiation. While loss of Cav1.2 does not affect motor learning, loss of Cav1.3 does impair motor learning. Our results help expand our understanding of LTCC function in cerebellar neurodevelopment and function.

OBJECTIVES/GOALS: Fatigue is a distressing side effect of cancer and its treatment. It is a subjective symptom that can include mental, physical, emotional, and motivational components. We sought to determine whether preventing inflammation affects fatigue-like behavior in a mouse model of radiation therapy. METHODS/STUDY POPULATION: C57BL/6 mice received three consecutive 8 gray doses of daily peripheral irradiation. We used voluntary wheel running activity to measure fatigue-like behavior before and after this period. Minocycline, an antibiotic with anti-inflammatory effects, was administered beginning a week before irradiation and continued throughout the experiment. We also tested mice lacking the toll-like receptor adaptor protein, MyD88. Cognitive abilities were tested using spontaneous alternation in a Y-maze. RESULTS/ANTICIPATED RESULTS: We found that minocycline reduces fatigue-like behavior exhibited after irradiation, but had no effect on pre-irradiation activity levels. Similarly, fatigue-like behavior after radiation was partially reversed by genetic loss of MyD88. Y-maze spontaneous alternation performance remained similar in all groups. DISCUSSION/SIGNIFICANCE OF IMPACT: Both pharmacological and genetic anti-inflammatory manipulations increased voluntary activity levels after irradiation. Our results suggest that inflammation is an important factor in the development of fatigue-like behavior. Modulators of inflammatory processes hold potential for alleviating fatigue.

OBJECTIVES/GOALS: Frequent utilizers of emergency departments (ED) make up a substantial share of overall ED use. Within pediatric emergency departments (PED), infants represent an age group that make up a disproportionate share of PED visits. The objective was to compare patterns of PED use for children less than 1 year of age by visit frequency and resource utilization. METHODS/STUDY POPULATION: Retrospective cohort study of infants less than 1 year presenting to 5 EDs in one health system over a 5-year period, with a 365-day follow-up after each index visit. Patient characteristics (age, sex, race/ethnicity, presence of chronic condition) and visit characteristics (arrival day/time, acuity level, disposition, testing (labs and radiographs, medications) were assessed. The relationship between patient and visit characteristics with utilization and repeat visits was assessed using multivariable regression. RESULTS/ANTICIPATED RESULTS: A total of 20,620 patients with 33,127 ED visits during study timeframe. Thirty three percent (n=6842) had more than one visit in a year; 3964 (19.2%) had two visits, 1542 (7.5%) had three visits, and 1336 (6.5%) had 4 or more visits. Across all visits, over half (52%) were low acuity. The most common diagnoses were respiratory diseases (27%), systemic states (including fever, viral illness, 23%), and gastrointestinal diseases (15%). These diagnoses remained the most common for those with 1, 2, 3, and ≥ 4 ED visits during follow up. As ED visit frequency increased, there was an increase in percentage of children who were older, non-Hispanic Black, and triaged as low acuity. Infants with ≥ 4 ED visits were more likely to be without a chronic condition, have no medications or testing ordered, and be discharged. DISCUSSION/SIGNIFICANCE: There was high ED utilization for those without chronic conditions who were least likely to need medications, testing, and hospital admission. With increasing attention paid to high-utilization in healthcare, it is important to assess why infants use the ED at high rates and develop systems to improve high value care while decreasing resource burden.

As the pace of biomedical innovation rapidly evolves, there is a need to train researchers to understand regulatory science challenges associated with clinical translation. We describe a pilot course aimed at addressing this need delivered jointly through the Mayo Clinic Center for Clinical and Translational Science and the Yale-Mayo Center for Excellence in Regulatory Science and Innovation. Course design was informed by the Association for Clinical and Translational Science’s Regulatory Science Working Group’s competencies. The course used didactic, case-, and problem-based learning sessions to expose students to regulatory science concepts. Course evaluation focused on student satisfaction and learning. A total of 25 students enrolled in the first two course deliveries. Students represented several disciplines and career stages, from predoctoral to faculty. Students reported learning “an incredible amount” (7/19, 36.8%) or “a lot” (9/19, 47.4%); this was reflected in individual coursework and their course evaluations. Qualitative feedback indicated that assignments that challenged them to apply the content to their own research were appreciated. The heterogeneity of students enrolled, coupled with assessments and course evaluations, supports the statement that there is a growing need and desire for regulatory science-focused curricula. Future research will determine the long-term impact.

Developing the translational research workforce is a goal established by the National Center for Advancing Translational Science for its network of Clinical and Translational Science Award Program hubs. We surveyed faculty and research staff at our institution about their needs and preferences, utilization of existing trainings, and barriers and facilitators to research training. A total of 545 (21.9%) faculty and staff responded to the survey and rated grant development, research project development, and professional development among their top areas for further training. Faculty prioritized statistical methods and dissemination and implementation, while staff prioritized research compliance and research administration. Faculty (73.9%; n = 119) and staff (87.3%; n = 165) reported that additional training would give them more confidence in completing their job responsibilities. Time and lack of awareness were the most common barriers to training. Our results indicate the value of training across a range of topics with unique needs for faculty and staff. This pre-COVID survey identified time, awareness, and access to training opportunities as key barriers for faculty and staff. The shift to remote work spurred by the pandemic has further heightened the need for effective and readily accessible online trainings to enable continuous development of the clinical and translational research workforce.

There is a growing trend for studies run by academic and nonprofit organizations to have regulatory submission requirements. As a result, there is greater reliance on REDCap, an electronic data capture (EDC) widely used by researchers in these organizations. This paper discusses the development and implementation of the Rapid Validation Process (RVP) developed by the REDCap Consortium, aimed at enhancing regulatory compliance and operational efficiency in response to the dynamic demands of modern clinical research. The RVP introduces a structured validation approach that categorizes REDCap functionalities, develops targeted validation tests, and applies structured and standardized testing syntax. This approach ensures that REDCap can meet regulatory standards while maintaining flexibility to adapt to new challenges. Results from the application of the RVP on recent successive REDCap software version releases illustrate significant improvements in testing efficiency and process optimization, demonstrating the project’s success in setting new benchmarks for EDC system validation. The project’s community-driven responsibility model fosters collaboration and knowledge sharing and enhances the overall resilience and adaptability of REDCap. As REDCap continues to evolve based on feedback from clinical trialists, the RVP ensures that REDCap remains a reliable and compliant tool, ready to meet regulatory and future operational challenges.