OBJECTIVES/GOALS: Participating in out-of-school activities (i.e., sports) is associated with improved physical and mental health, but racial/ethnic and income disparities persist. Our goal was to describe parent perspectives to understand how to enhance family engagement in these activities, especially in low-income Black and Latinx communities. METHODS/STUDY POPULATION: We recruited parents through a large urban parks and recreation department located in Los Angeles County (LAC) and two community-based organizations who provide services primarily to low-income Black and Latinx residents in LAC. We conducted semi-structed English or Spanish interviews with parents of children ages 6-17 about the impact of activities on families, experiences making it difficult or easier for their child to be involved, and recommendations to meet their needs and facilitate family engagement. We coded the interviews using Dedoose and then performed thematic analysis. RESULTS/ANTICIPATED RESULTS: We completed 34 interviews (17 English, 17 Spanish). Parents shared these central ideas about out-of-school activities: (1) they support families and promote healthy child development; (2) they must create a safe environment and promote a sense of belonging for all families; and (3) service delivery changes reduced barriers, but activities often remained inaccessible. Barriers included lack of awareness about low-cost options, online enrollment, financial costs, waitlists, program timing, inaccessible parks/pools, safety, inconsistent quality, few opportunities for families to participate together, and a desire for more inclusive practices around race, culture, language, gender, children with special needs, and parent involvement. Parents offered recommendations to increase family engagement. DISCUSSION/SIGNIFICANCE: Involving parents in optimizing out-of-school activities may improve access, uptake, and quality of these health-promoting activities in low-income Black and Latinx communities.

OBJECTIVES/GOALS: The objectives of the ICPC Course were 1) to define elements needed to 'build' a solid framework for the creation of an effective informed consent document; 2) to examine various regulatory references and their importance in the context of the informed consent process; and 3) to review helpful tips and best practices of the informed consent process. METHODS/STUDY POPULATION: This 7-week course was a multi-disciplinary effort by the Miami CTSI, the Regulatory and Monitoring Support Office and the Human Subjects Research Office (HSRO), as part of their co-op partnership. The CTSI’s Network of Clinical Research Professionals (NCRP) (over 1995 members and established to offer and enhance education, training, and overall career development and networking for the U’s diverse research workforce) served as the target audience. The lead HSRO official was the course content developer and served as instructor. The Director of the HSRO served as the certificate & program coordinator. Additionally, this course indirectly fulfilled part of the requirements of the Institution’s AAHRPP re-accreditation. RESULTS/ANTICIPATED RESULTS: 70 persons attended all sessions and received their certificate. A post-course survey was deployed (n=70). 69 respondents were extremely satisfied with course content; 67 were likely to recommend the course to a colleague. Feedback highlights include: appreciation for time taken to organize/offer the course; presenter knowledge and preparation; and that case studies/examples were up-to-date and applicable. As an added incentive to participate in the course, both the School of Nursing and the Medical School provided CEU credits and the Society of Clinical Research Associates (SOCRA) accepted documentation of candidate participation in continuing education programs for recertification. DISCUSSION/SIGNIFICANCE: Based on a need identified through a multi-disciplinary process, a successful Informed Consent Professional Certificate Course was created and implemented. Respondents indicate extremely high satisfaction with the course and felt it was very relevant and applicable to their practice.

OBJECTIVES/GOALS: Prostate cancer treatment is associated with significant genitourinary side effects. There is a critical need for treatment with decreased morbidity. We report the development of a novel treatment paradigm combining irreversible electroporation and lower dose radiation to provide prostate cancer patients with a less morbid treatment. METHODS/STUDY POPULATION: Intermediate risk prostate cancer patients will undergo focal irreversible electroporation followed by low dose, whole gland radiation therapy. The primary endpoint is freedom from clinically significant cancer on biopsy at 12-month follow up. Secondary endpoints include safety profile, oncologic efficacy, effectiveness of RT and need for secondary treatment. This trial (NCT05345444) and currently actively recruiting patients after initial feasibility trial. Sample size is calculated to detect an increase in the proportion of patients who are cancer free at 1-year, from 0.80 to 0.95. An exact binomial test with a 10% one-sided significance level will have 94.3% power to detect the difference between the null and alternative hypothesis when the sample size is 42. RESULTS/ANTICIPATED RESULTS: This is a clinical trial in progress. DISCUSSION/SIGNIFICANCE: Combined irreversible electroporation (IRE) and a lower dose radiotherapy (RTIRE) may provide prostate cancer patients a treatment with minimal side effects.

OBJECTIVES/GOALS: We prioritize Chlamydia’s public health impact, aim to develop rVCG-MECA for practical use, study robust immunity for effective strategies, and assess animal models for human vaccination adaptation. Our work highlights rVCG-MECA’s translational significance in public health. METHODS/STUDY POPULATION: Female Mice C57BL/6J mice (N=8) were immunized intramuscularly(IM) and intranasally(IN) and boosted twice, two weeks apart, with rVCG-MECA, once with live Chlamydia (C. trachomatis serovar D elementary bodies) and PBS. Specific mucosal and systemic immune responses were characterized. Vaccine efficacy was determined from chlamydia shedding following the transcervical challenge. Additionally, Chlamydia-specific cytokine (IFN-γ and IL-4) production by splenic and ILN T cells was assessed after 16 weeks RESULTS/ANTICIPATED RESULTS: Immunization with rVCG-MECA via intramuscular and intranasal routes triggered notable humoral responses in systemic and mucosal tissues. Intramuscular vaccination produced higher IgG2c levels in both tissues, while intranasal vaccination led to elevated IgA levels in mucosal tissues. rVCG-MECA-immunized mice exhibited significantly higher IFN-γ (Th1) secretion compared to IL-4 (Th2), with intramuscular immunization showing the highest IFN-γ levels. These findings anticipate robust immune responses, promising protection against Chlamydia, particularly through the intra muscular route. Overall, our results support rVCG-MECA as a promising Chlamydia vaccine, aligned with public health goals. DISCUSSION/SIGNIFICANCE: This study suggests that IM and IN immunization with rVCG-MECA induces immune effectors such as IFN-gamma and IgG2c that mediate chlamydial clearance in the genetical tract.

OBJECTIVES/GOALS: The CTSI Research Infrastructure Network (RIN) expands CTSI’s reach into the regional health systems to provide our services to a broad and diverse translational science community. We create and support research collaborations that span multiple geographies and patient populations and serve as a bridge between the affiliate sites and CTSI programs. METHODS/STUDY POPULATION: We conducted needs assessments at each site (n=6) via in-depth, semi-structured interviews with key stakeholders. Informants (n=40) included investigators, study personnel, and research administrators. Investigators were selected across a variety of departments and career stages. Interview transcripts and notes were analyzed using matrix-based qualitative methods to identify both the common and unique research infrastructure needs of each site. Individualized support plans were shared with each site and a comprehensive summary report was presented to CTSI leadership. RIN met with UCSF’s Comprehensive Cancer Center, which conducts clinical trials at 2 sites, to coordinator our effort and services. When possible, RIN addressed service requests in real time that arose during interviews. RESULTS/ANTICIPATED RESULTS: We identified heterogeneous needs across multiple sites. However, among the community health systems with non-academic clinicians, there were common needs for research training, consultations in biostatistics/study design, and finding academic collaborators. The needs of sites with UCSF academic faculty differed from those of community sites and mainly included improved awareness and access to CTSI programs, ease of use of data extraction services, training programs, and assistance with regulatory approvals. Site needs are best addressed with individual plans created with CTSI Program leaders. A developing governance structure will include representation on a CTSI advisory committee and an annual conference to facilitate the sharing of best practices and foster collaboration across member sites. DISCUSSION/SIGNIFICANCE: Providing an individualized, site-specific approach to expanding CTSI services to regional health systems, will increase research collaborations across Northern California through building relationships, addressing unique infrastructure needs and sharing best practices throughout the network.

OBJECTIVES/GOALS: Clinical Issue/ Practice Problem: A high turnover rate for research nurses was identified between 2017-2022 in the clinical research center at an urban Midwest academic health center. Inexperienced staff and high turnover are barriers to maintaining high-quality research integrity, efficacy, and safety for research projects and participants. METHODS/STUDY POPULATION: Project Implementation: A formal mentorship program was developed based on a curriculum from the International Association of Clinical Research Nursing Scope and Standards of Practice. The six-week project was implemented for research nurses with less than 2 years of experience. Mentees were paired with senior research nurses and met one-on-one weekly. Mentees completed the Anticipated Turnover Scale (ATS) survey in week 1 and week 6. All program participants completed a final evaluation survey. RESULTS/ANTICIPATED RESULTS: Outcomes: There was a one-point average reduction in pre- and post-ATS survey scores. This result supports the theory that mentees were less likely to leave their research role after a formal mentorship program. Qualitative results from the final evaluation survey demonstrate the program had a positive impact and benefits for both the mentees and the mentors. DISCUSSION/SIGNIFICANCE: Clinical Implications: By decreasing turnover rates, a highly competent and knowledgeable research nursing staff is attained to ensure appropriate nursing action and safety profiles for novel therapies.

OBJECTIVES/GOALS: Disparities persist among minority and marginalized populations in immunizations and research. The Clinical Research Vehicle Team aimed to bring research to our community’s doorstep, building generalizability of research outcomes. Our mission adapted due to the pandemic to include increasing COVID-19 vaccinations and health education. METHODS/STUDY POPULATION: Community Engagement Events are planned when event organizers submit a request, then the proposal is discussed by the Clinical Research Vehicle (CRV) Team and UF HealthStreet. We go to underserved communities throughout Florida, food distributions, churches, community centers, rural areas, etc. At these events we offer services at no cost, including: COVID-19 and flu vaccines, as well as health screenings. We provide health education to individuals and refer them to UF HealthStreet for community resources, as appropriate. Research events are planned when researchers submit a request for the CRV, then the request is discussed by the CRV Team. The studies covered topics from COVID-19 research to the development of a diverse health database. Our fleet consists of two Community Health Vehicles and the CRV. RESULTS/ANTICIPATED RESULTS: As Community Engagement Events are ongoing, the presented numbers are reflective of the data available at the time of submission. Our impact includes administering 1,606+ COVID vaccines, conducting 1,081+ health screenings, collecting data from 944 screenings, providing 265 Flu vaccines, supporting 267+ community health events, and serving 25+ communities. The Clinical Research Vehicle facilitated 6 IRB-approved studies, administration of monoclonal antibody infusions, extended the capabilities of the Clinical Research Center, and enabled clinical trials aimed for novel treatments for COVID-19 positive participants. DISCUSSION/SIGNIFICANCE: Championing this initiative has shown an impact on vaccination rates and health education in communities. Experience and knowledge gained by our team throughout the pandemic while engaging with community members provided a platform from which to build generalizability of research outcomes and support translational science.

OBJECTIVES/GOALS: A Mayo Clinic in Florida committee completes 100+ Scientific Reviews annually through manual e-mail and Excel tracking, placing a manual burden on reviewers and coordinators. REDCap, an electronic data capture system, was leveraged to reduce the administrative burden.#_ftn1 #_ftnref2 METHODS/STUDY POPULATION: Historically, emails were sent by a coordinator to physicians, requesting their initial review and following up with reminders. This process was tracked using Excel, presenting a need to make this process more efficient, so a workgroup was created. To ensure all perspectives were accounted for, the workgroup included the review coordinator, a physician reviewer, and study team members who submit development requests for studies. The process was mapped using the existing Excel spreadsheet, and email templates. Critical data elements were identified, ensuring the database would identify bottlenecks. Two REDCap instruments were then created: one to outline the coordinator workflow and a survey for physician reviewers to complete the scientific reviews. #_ftn2#_ftnref1 RESULTS/ANTICIPATED RESULTS: The workflow is live in REDCap and has effectively processed over 100 scientific reviews in <1 year. The system captures the review status and guides the coordinator through the workflow, capturing dates when tasks are completed. When review criteria are met, the database sends an email to the assigned reviewer. The email includes a link to a REDCap survey, containing all pertinent documents. The reviewer uploads their completed review form within the survey, if this is not completed within a given period, the database sends email reminders. Once the review is complete, a notification is sent to the review coordinator. The review workflow is accessible to the study team who requested scientific review, making them aware of their request status ad giving them access to the review the moment it is completed. DISCUSSION/SIGNIFICANCE: Leveraging REDCap has increased visibility, reduced overall manual processes, and simplified the reviewer burden by providing all the information needed in a single notification. REDCap is a cost-effective, impactful solution to simplifying administrative burden in managing committee reviews.

OBJECTIVES/GOALS: Investigators are looking to integrate DE&I into trials, increasing study population diversity. In response, the Community and Recruitment programs of the Oregon Clinical and Translational Research Institute (OCTRI), built a collaborative, community-focused system for investigators interested in community engagement and recruitment. METHODS/STUDY POPULATION: Historically, the OCTRI Community and Recruitment programs operated independently. To build a community-focused support system for investigators, we began with the programs learning about each other’s goals, values, and operations over a six-month period. Over the next two years, we then began integrating into each other’s team meetings and processes. Each team has their own process for providing services to investigators, consisting of intake and consultations. To these, we added tags for the other’s program topics. This allowed efficient identification of opportunities for referrals and joint consultations. Program leads meet regularly to discuss referrals and partnerships allowing for more effective and consistent collaboration. RESULTS/ANTICIPATED RESULTS: Over the last two years, OCTRI Community and Recruitment built a collaborative process that allows investigators to work with both programs, receiving comprehensive, supportive assistance and education on how to design their study, engage with community, and recruit participants in a community-based setting. During this time, the programs have: provided consultations that discussed the other’s program (N=21); made direct referrals to the other program (N=4); and, provided joint consultations (N=2). Studies that are investigator-initiated, in planning phase, and population focused were found to benefit most from the collaborative efforts. Additionally, we have improved the collaborative process during this time, resulting in more streamlined, efficient support services for investigators. DISCUSSION/SIGNIFICANCE: Diverse study populations and DE&I are essential to research. This collaborative, community-engaged recruitment approach by our programs allows investigators to leverage both programs’ resources and expertise. We continue to identify best practices and study types that benefit from this collaborative approach to improve our services.

OBJECTIVES/GOALS: Human-centered design (HCD) provides a practical, collaborative approach to integrating diverse perspectives throughout the translational research process. We highlight two CTSAs with established HCD teams who are utilizing varied approaches to accelerate the use of HCD in health intervention design and community engagement. METHODS/STUDY POPULATION: The CTSAs at the University of Michigan (MICHR) and the University of Pittsburgh (Pitt CTSI) have dedicated teams of human-centered designers that assist investigators with the application of HCD in their studies. MICHR’s service approach utilizes facilitated Design Sprints with study teams, guiding them from early conception of a research question through the co-design of interventions and innovations with end-users. Pitt CTSI’s training and consultation approach employs a two-day intensive training with group coaching sessions that provide investigators and research staff with skills and knowledge needed to implement HCD within translational research projects. Both hubs offer consultations on methodology for investigators interested in pursuing funding for studies utilizing HCD. RESULTS/ANTICIPATED RESULTS: Research teams at both hubs are pursuing HCD to co-design health research interventions and mHealth technologies with end-users locally and internationally, to facilitate meaningful engagement within advisory boards and collaboratives, and to enhance team science. To date, MICHR has conducted 13 research study-focused Design Sprints with 73 participants, as well as providing consultations to 26 research teams. Pitt CTSI has trained 164 investigators and staff and provided 146 consultations with research teams. Requests for HCD training have increased at MICHR while requests for HCD service provision have increased at Pitt CTSI. Both hubs are now building capacity to enact a more holistic approach to HCD that combines service delivery and training approaches to better meet the needs of investigators. DISCUSSION/SIGNIFICANCE: Increasing CTSA hub capacity to support the use of HCD in translational research by offering service and training opportunities can position investigators to conduct high impact health research that elevates the voices of those most likely to benefit from interventions, treatments, and innovations.

OBJECTIVES/GOALS: Maternal opioid use disorder (OUD) is linked to poor fetal outcomes. While it has been established that opioids can cross from maternal to fetal circulation, the mechanisms underlying these adverse outcomes remain poorly defined. This study aims to uncover OUD-associated immunological changes in maternal and fetal circulation. METHODS/STUDY POPULATION: To study the effect of maternal OUD on the maternal immune system at delivery, we collected maternal blood samples at delivery from healthy pregnant women (controls) and pregnant women with a diagnosis of severe OUD. To study the impact of maternal OUD on newborn immunity, we also collected umbilical cord blood (UCB) at delivery. Flow cytometry was used to determine the frequency and phenotype of circulating immune cell subsets and responses to stimulation. Isolated monocytes were stimulated with bacterial/viral agonist cocktails, while T and NK cells were stimulated with PMA/ionomycin. The impact of maternal OUD on circulating immune mediators was determined by Luminex and on monocyte activation markers sCD14 and CRP by ELISA. RESULTS/ANTICIPATED RESULTS: In maternal circulation, OUD was associated with a significant decrease in markers of inflammation, cell proliferation, and activation. Frequencies of immune cell subsets were impacted by OUD, shown by an expansion of CD8+ EMRA T cells, marginal-zone B cells, mDCs, non-classical monocytes, and CD16low NK cells. While no differences were seen in T and NK cell responses to stimulation, monocytes and pDCs had significantly lower responses to bacterial and viral agonist stimulation. Analysis of UCB revealed increased levels of pro-apoptotic/T cell exhaustion mediators and pro-inflammatory cytokines, albeit decreased levels of several chemokines and growth factors. The UCB immune landscape is altered with maternal OUD, as demonstrated by a shift from naive to memory CD8+ T cells and a decrease in pDC frequency. DISCUSSION/SIGNIFICANCE: OUD dampens maternal peripheral immunity, possibly contributing to poor placental function or premature/delayed labor. Monocytes and pDCs lack antimicrobial functionality, suggesting increased infection susceptibility with OUD. Finally, these implications extend to the fetal compartment, shown by heightened immune activation in UCB.

OBJECTIVES/GOALS: The objective is to describe the process for developing two measurement tools to measure confidence (self-efficacy) of the anti-racist advocate in an academic setting. METHODS/STUDY POPULATION: We proposed five spheres encountered by the academic trainees: Academic/Research, Clinical, Policy, Interpersonal, and Intrapersonal. We evaluated a book, by Shereen Daniels, used in anti-racism literature: The Anti-Racist Organization - Dismantling Systemic Racism in the Workplace. Using the proposed metric of RACE framework, Recognize the problem, Analyze the impact, Commit to action, Empower for change, we sought to establish readiness on the spectrum of anti-racism advocacy. We developed a list of anti-racism and anti-bias advocacy skills based on: 1) Informational interviews with anti-racism and anti-bias experts, 2) Scoping literature review and 3) Academic trainees’ and faculty lived experience. RESULTS/ANTICIPATED RESULTS: The first assessment, “5-Spheres”, consists of 10 items that perform, 1) Analysis of readiness on the spectrum of anti-racism advocacy using RACE framework (Figure 1 [https://drive.google.com/file/d/1A3nMArEn7ZSxZSuSgDkYl_row-VOhOXf/view?usp=drive_link]), 2) Assessment of workplace environment. The second assessment, “Skills”, consists of 25 items (Figure 2 [https://drive.google.com/file/d/1GTdfSgn0-mPu-flSUVSN-vIKTxBCkFW3/view?usp=drive_link]) that perform assessment of confidence of specific skills within each of the five spheres using the following scale: 1 – Not confident at all, 2 – Lacking some confidence, 3 – Somewhat confident, 4 – Completely confident DISCUSSION/SIGNIFICANCE: This proposed measurement tool can extend to anti-bias as well as anti-racism. Potential uses of the self-assessment includes: 1) Measurement and 2) Gap-spotting.

OBJECTIVES/GOALS: Our project aims to assess the composition or characteristics of research papers that score high on alternative metrics. These alternative metrics including the number of newspaper mentions, social media mentions, and the attention score as catalogued on Altmetric, a tool used to document community attention for a given research paper. METHODS/STUDY POPULATION: Our study intends to 1) Utilize topic modeling to identify prevalent themes on Altmetric, and 2) Apply network analysis to elucidate the interconnectedness among universities, funding sources, journals, and publishers associated with high-attention papers. 3) Examine how these patterns vary when attention metrics shift, such as social media mentions, newspaper mentions, or the Altmetric score. We'll first perform this analysis on all types of papers and then limit the networks to Biomedical and Clinical Sciences, and Public and Allied Health Sciences to help inform what health topics garner attention. RESULTS/ANTICIPATED RESULTS: Our initial Altmetric topic models revealed sustained attention for COVID-19 and vaccination-related publications well beyond the pandemic (specifically, papers from January 2023). Health topics like cancer, dementia, and obesity also garnered high attention. Additionally, political papers (elections, democracy), climate change, and battery research had notable attention values. Further analysis needs to be done to explain why these topics gain attention and the type of attention they garner. We will construct networks to see the relationship between attention and entities like universities, funding sources, journals, and publishers. This will identify whether certain clusters of these entities produce papers with high attention or if attention is distributed evenly amoung them. DISCUSSION/SIGNIFICANCE: To gauge the broader impact of scholarly research alternative metrics beyond citations are needed. Altmetric is used widely by CTSA’s to measure the community interest in research. Understanding the types of research that gain traction on Altmetric can help researchers understand how to garner interest from the community.

OBJECTIVES/GOALS: With a growing intеrеst in tailoring disеasе diagnosis to еach individual as opposеd to a “onе-sizе-fits-all” approach, our aim is to еnhancе thе robustnеss of thе Itеrativе Clustеring for Subtypе Discovеry (iKCAT) algorithm in charactеrizing lung cancеr subtypеs for individualizеd trеatmеnt. METHODS/STUDY POPULATION: Our mеthod еxplorеs thе robustnеss of thе prеviously dеvеlopеd iKCAT algorithm. This itеrativе clustеring mеthod finds robust—homogеnеous and diffеrеntiablе—subtypеs of lung nodulеs through itеrativе K-mеans clustеring that hеlps classify thеm and lеavеs somе data unclustеrеd. This sеt of unclustеrеd or “hard” data rеprеsеnts imagеs that cannot confidеntly bе assignеd to any subtypеs and may rеquirе morе rеsourcеs (е.g., timе or radiologists) to diagnosе. Wе еxplorе thе robustnеss of iKCAT across multiplе fеaturе spacеs, including dеsignеd imagе fеaturеs (which arе еnginееrеd to capturе somе propеrtiеs such as lеvеl of еlongation, еccеntricity and circularity), rеducеd dеsignеd imagе fеaturеs using Principal Componеnt Analysis (PCA) and Uniform Manifold Approximation and Projеction (UMAP). RESULTS/ANTICIPATED RESULTS: Whеn running our еxpеrimеnt on thе 64 imagе fеaturеs, our rеsults consistеntly carvеd out a singlе purе, homogеnеous clustеr ovеr thе coursе of 30 iKCAT runs. From an initial datasеt of 1490 data points, 1430 points wеrе lеft unclustеrеd in this fеaturе spacе. Whеn conducting thе 30 iKCAT runs on thе PCA fеaturе spacе with 10 componеnts, wе found it did not producе any distinct clustеr abovе thе dеfinеd homogеnеity thrеshold. Thе 2D UMAP fеaturе spacе consistеntly gеnеratеd 8 clustеrs with an avеragе homogеnеity of 87. 22% ovеr 30 runs, and only lеft 9 points unclustеrеd. Ovеr 30 iKCAT runs, wе idеntifiеd 8 pеrsistеnt clustеrs or subtypеs, 3 mostly malignant and 5 mostly bеnign clustеrs. DISCUSSION/SIGNIFICANCE: Through our еxpеrimеnt using thе iKCAT algorithm, wе found that iKCAT’s clustеring functionality producеd thе most pеrsistеnt rеsults on thе 2D UMAP fеaturе spacе duе to its high avеragе homogеnеity scorеs and consistеncy in idеntifying clustеrs/subtypеs, hеlping improvе tailorеd disеasе diagnosis.

OBJECTIVES/GOALS: We will reduce the number of research staff by 50% within 6 months that are non-compliant with completing the Good Clinical Practice (GCP) course without increasing costs as measured by salary and benefits for staff involved in the tracking and assignment process. We will also introduce a GCP policy to define the education requirements for GCP training METHODS/STUDY POPULATION: We touted focus groups to stakeholders for input on workflows with graphs, surveys, and meetings in the onset and throughout the project. This input prompted us to develop a staff guide outlining the modification process of removing a staff name from an IRB trial if they are no longer active in the trial. A workflow and root cause analysis were done resulting in the implementation of one gatekeeper for assigning and tracking completions, instead of three staff. Successfully reconciling the non-compliant report, which extracted data from three databases, allowed us to eliminate 174 names from the report, which originally comprised 792 names. IRB modifications were entered into the IRB system for these 174 names. We also put into effect a GCP policy for the institution, where none had previously existed. RESULTS/ANTICIPATED RESULTS: * In 3.5 months, we decreased the GCP non-compliance rate by 50% from 792 to 399. * We removed 22% of names from the 792 from the report due to their status of being non-employees, or Emeritus, or not in research anymore. * We discovered a data type issue in the non-compliance report that shuffled the MAX calculation, therefore not requiring some staff to complete the training. * We developed a new process for assigning training, resulting in faster compliance rates for the institution. It included sending emails to users two months before their training expired before we assigned the course to them. * We reconciled the non-compliance report, and it decreased the effort with staff involved in GCP for grant renewals and audits in numerous other departments. * Developed an escalation procedure for non-compliant staff. DISCUSSION/SIGNIFICANCE: Failure to address our GCP non-compliance rates could have put our institution at risk for potential penalties. With 4400+ research personnel listed on active trials the interventions we implemented accelerated our compliance rate by assigning the refresher course monthly and this also resulted in no disruption to staff completing the training.

OBJECTIVES/GOALS: Limited access to medication and poor medication adherence exacerbate chronic diseases following disasters. Experts recommend individuals in disaster-prone areas be prepared to manage their chronic diseases in the event of resource disruption. This study’s goal is to identify factors underlying personal medical preparedness. METHODS/STUDY POPULATION: A cross-sectional survey of 120 insured adults age ≥50 in Southeast Louisiana with hypertension and ≥1 daily medication during the 2023 Atlantic Hurricane Season is underway. The survey includes the Household Emergency Preparedness Index Access and Functional Needs Section (HEPI AFN), a validated measure of medical preparedness that accounts for special circumstances including refrigerated medication and electricity-dependent medical equipment. The mean score of the 9-item tool ranges from 0 to 1, with higher scores indicating more preparedness. The survey also includes 3 open-ended questions where participants can explain difficulties with medication adherence during hurricanes in their own voice. Data collection is ongoing. This interim analysis provides descriptive statistics. RESULTS/ANTICIPATED RESULTS: An interim analysis of the first 50 respondents included 46% women, 52% Black, mean age 61.2 (SD=7.3) years, and mean 52.5 (SD=16.2) years living in a hurricane-impacted area. Participants had a median of 1 comorbid condition; 72% reported taking >5 daily medications. Most respondents (94%) stated their household was at least “somewhat prepared” to handle a disaster and reported medical preparedness on an average of 82% of HEPI-AFN items (mean score = 0.82, SD=0.18); 90% reported that they had never had a healthcare worker talk to them about personal medical preparedness. On open response questions, participants cited insurance restrictions as the primary barrier to having extra medication on hand. In the final sample, regression models will be used to examine factors associated with increased preparedness. DISCUSSION/SIGNIFICANCE: While most participants in this insured, disaster-experienced preliminary sample are medically prepared, few have discussed preparedness with a healthcare provider. Identifying socio-demographic factors associated with preparedness will help to strengthen mitigation strategies addressing widening of health disparities during disasters.

OBJECTIVES/GOALS: Women are twice as susceptible to ailments like anxiety, PTSD, and Alzheimer’s disease compared to men. The locus coeruleus (LC), the primary source of noradrenaline for the brain is implicated in these disorders, however physiological sex differences have never been assessed in the LC. METHODS/STUDY POPULATION: To address this gap, In vivo electrophysiology under anesthesia was used to measure single unit activity of noradrenergic LC firing patterns in 4-month-old wild-type Fischer male and female rats. Recordings measured neuronal activity under basal conditions and in response to a footshock stimulation which elicits burst firing in LC neurons. Single unit activity is sorted via automatic valley seeking scan PCA, additional manual sorting is done via line and template method. Analysis is done extracting interspike interval (ISI) and firing rate of single units, additional analysis is done to quantify properties of bursting patterns (burst duration, spikes per burst, interburst interval, etc…). RESULTS/ANTICIPATED RESULTS: This data shows that during LC burst firing, females have longer interspike intervals compared to males, supporting the inhibitory effect of E2 on LC firing. Additionally, females have significantly different waveform patterns than males, indicating possible differences in intrinsic properties, but further supporting sexually distinct physiology of the LC . Because female rats have been estrous cycle tracked via vaginal lavage, stratification into estrus groups and further analysis may uncover differences within females. These data suggest that estrogen acts as a potent neuromodulator of noradrenergic LC neurons, providing valuable insights into the physiology of this brain region. DISCUSSION/SIGNIFICANCE: This study is the first exploration of LC physiological sex differences. This work offers insights into a critical brain region implicated in many diseases, and may pave the way for future therapeutic approaches, particularly for women, who are at a higher risk of neurological disease developing.

OBJECTIVES/GOALS: 48,000,000 people in the U.S. have hearing loss, negatively impacting quality of life and work. Unveiling axon guidance for auditory type II spiral ganglia neurons (SGNs) will aid development of new therapies. I study the role of Eph/Ephrin and planar cell polarity (PCP) signaling during type II SGN turning and outer hair cell (OHC) innervation. METHODS/STUDY POPULATION: This quantitative study was conducted on Efna3 and Vangl2 null mice possessing Neurog1CreERT2 and R26RtdTomato mutations. Spontaneous Cre activity within the Neurogenin1CreERT2 line causes recombination and expression of fluorescent Rosa26 Reporter (R26R)tdTomatoin a restricted number of SGNs, including type IIs. Together, these lines permit SGN sparse labeling. Immunostaining and confocal imaging were used to analyze dsRed in Efna3 and Vangl2 mice and quantify type II SGN turning. In combination, Imaris 3D renderings were used to quantify type II SGN turning, branching, navigation features and temporal effects of EPHRIN-A3-Fc on type IIs via cochlear cultures (a gain-of-function manipulation). For both sexes, 5-6 cochleae per genotype were analyzed and compared by t-test to wildtype (WT) controls. RESULTS/ANTICIPATED RESULTS: Efna3 nulls showed a small rise in type II SGNs incorrectly turning toward the apex at an error rate of 16.0% compared to WTs (n=6; p=0.05). P0 Efna3 nulls had reduced branch number compared to WTs, 4.1 and 7.2, respectively (n=129; p=<0.0001), suggesting EPHRIN-A3 acts as a positive growth cue. In cochlear cultures, EPHRIN-A3-Fc led to type II SGN collapse at E15.5, indicating a repulsive function. However, at P0, EPHRIN-A3-Fc treatment led to type II SGNs with elevated branch numbers compared to Control-Fc treatment: 18.1 and 11.4, respectively (n=116; p=<0.0001). This indicates a positive growth function. At E16.5, EPHRIN-A3 protein immunoreactivity on Deiters’ and pillar cells appears reduced in Vangl2 nulls compared to WT cochleae, suggesting that EPHRIN-A3 acts downstream of PCP signaling. DISCUSSION/SIGNIFICANCE: Results suggest that Eph/Ephrin signaling acts downstream of PCP signaling to mediate type II SGN guidance and EPHRIN-A3 switches its mode of activation. The clinical implications of these findings are that therapeutics targeting EPHRIN-A3 and/or VANGL2 in their given pathways could stimulate new OHC innervation following auditory damage.

OBJECTIVES/GOALS: The dominant complication of Systemic Sclerosis (SSc) is clinically severe and commonly fatal pulmonary fibrosis (PF). We sought to determine the downstream regulatory role of the basic Helix-Loop-Helix protein 40 (bHLHe40), in response to Insulin-like Growth Factor II (IGF-II) on Pro-Lysyl Oxidase cleavage products. METHODS/STUDY POPULATION: We examined the response of primary pulmonary fibroblasts cultured from the lungs of control donors and SSc lung explants to IGF-II as well as human recombinant Lysyl Oxidase Propeptide (LOX-PP). In addition, we utilized an experimentally-induced model of lung fibrosis with intratracheal bleomycin administration. We used qPCR and immunoblotting to quantify mRNA and protein levels, respectively. We used sequence-specific small-interfering RNA to silence targeted genes. Immunoblots were quantified in ImageJ (NIH) and statistical analyses were performed in GraphPad Prism. RESULTS/ANTICIPATED RESULTS: IGF-II regulates levels of Pro-LOX, active LOX, and LOX-PP, as well as isoforms of proteases Bone Morphogenetic Protein 1 (BMP1) and Tolloid-like 1 (TLL1). The transcription factorbHLHe40 localizes to the nucleus in response to IGF-II. bHLHe40 silencing downregulated TLL1, abrogating the enzymatic cleavage of Pro-LOX. SSc lungs have higher baseline levels of the total (N-glycosylated/unglycosylated) LOX-PP than normal lung tissues, and baseline levels of LOX-PP correlated with TLL1 Isoform 2 in SSc lungs. LOX-PP contributes to the development and progression of SSc-PF by mediating changes consistent with the extracellular matrix deregulation implicated in SSc-PF: elevated levels of Collagen 3A1 (COL3A1), Fibronectin-1 (FN1), and Plasminogen Activator Inhibitor-1 (PAI1). DISCUSSION/SIGNIFICANCE: Our findings indicate that bHLHe40, TLL1, and LOX-PP may serve as targets of therapeutic intervention to stop the progression of SSc-PF. Since activation of common fibrotic pathways are involved in different diseases characterized by lung fibrosis such as IPF, our findings may have wider implications for lung fibrosis associated with other diseases.

OBJECTIVES/GOALS: This project focuses on investigating the potential of Simalikalactone D (SKD) as an anticancer agent, exploring the mechanisms underlying SKD’s induction of cell death, and assessing the impact of SKD on diverse breast cancer cell lines. Also, it Investigates the compound’s mechanisms of action beyond caspase 3-dependent pathways. METHODS/STUDY POPULATION: Three breast cancer cell lines were used: SKBR3, MDA-MB-231, and MDA-MB-468. Two triple-negative breast cancer cell lines are included to address cancer disparities across diverse ethnic backgrounds. Viability assays were conducted to determine half-maximal inhibitory concentrations (IC50). Caspase 3 activity assay was performed to evaluate apoptosis as a possible cell death pathway. Wound healing and colony formation assays are used to assess cell migration and clonogenic capacity. Proteomic analysis and phosphoarray analysis are planned for a deeper understanding of SKD’s anticancer properties, as well as testing for caspase 3 independent pathways. RESULTS/ANTICIPATED RESULTS: SKD demonstrated substantial cytotoxicity against all three breast cancer cell lines. IC50 values for SKBR3, MDA-MB-231, and MDA-MB-468 were 60.0 nM, 65.0 nM, and 116 nM, respectively. SKD induces cell death via caspase 3-independent pathways. Further experiments are needed to confirm and elucidate the molecular pathways being impacted. SKD inhibited cancer cell migration and clonogenic potential, suggesting it can reduce tumor growth and metastatic tendencies. DISCUSSION/SIGNIFICANCE: The study highlights SKD’s cytotoxicity across diverse breast cancer cell lines. It underscores the mechanism of action, a caspase 3 independent pathway. These findings hold promise for the development of innovative anticancer treatments and emphasize the importance of exploring varied cellular responses to mitigate global cancer disparities.