OBJECTIVES/SPECIFIC AIMS: Assess the presence of contact activation at baseline in sera from common variable immunodeficiency (CVID) patients with and without inflammatory complications compared with healthy controls. METHODS/STUDY POPULATION: CVID patients were recruited in the outpatient setting and the measurement of cleaved plasma HK (cHK) levels was determined by Western blot analysis, under reducing conditions, with quantitation of total and cHK bands using an Odyssey imaging system (Licor). One-way ANOVA test for differences among the 3 studied groups will be applied. Biomarkers C3, C4, C1 inhibitor levels and hs-CRP were also measured. RESULTS/ANTICIPATED RESULTS: Participant enrollment continues and to date, 9 CVID patients were studied, 7 with and 2 without inflammatory complications. Repeated determinations of cleaved HK% (cHK%) revealed an average of 1.20% (range: 0.46%–2.66%) in CVID patients with inflammatory complications and those without complications averaged 1.07% (range: 0.79%–1.35%). Healthy controls had an average cHK of 1.15% (range: 0.60%–2.10%). DISCUSSION/SIGNIFICANCE OF IMPACT: Cleaved kininogen detected in the sera of CVID patients was found at similar levels compared with healthy controls (cHK<5%). Findings suggest that systemic activation of the contact system might be absent in CVID, however, future considerations include developing detection methods for local tissue activation.

OBJECTIVES/SPECIFIC AIMS: The objective of this study is the pharmacology of sublingual Buprenorphine in Hispanics/Latino men and women. Specifically we plan to: (1) Administer sublingual buprenorphine to Hispanic/Latino men and women volunteers, and measure the circulating amounts of the drug in the bloodstream as a function of time; that is, pharmacokinetics of buprenorphine. The goal of the proposed study is to evidence that there are gender and ethnic differences in the pharmacokinetics of sublingual buprenorphine between not only Hispanics/Latinos and non-Hispanics/Latinos (Caucasian), but also within Hispanic/Latino men and women. METHODS/STUDY POPULATION: We are proposing a phase 1 of buprenorphine using 12 healthy volunteers. To test for differences in pharmacokinetics between Hispanic/Latino men and women, 6 Hispanic/Latino men, and 6 Hispanic/Latino women 21 years of age and older will be recruited. The volunteers should be living in Puerto Rico, and must have both parents born in Puerto Rico. Sublingual buprenorphine will be administered using a low dose of 16 mg one time only. Blood samples will be collected from each volunteer at t=0, 1, 2, 4, 6, 8, 12, and 24 hours after administration. The amount of circulating drug in the bloodstream of the volunteers will be measured using liquid chromatography combined with mass spectrometry. Pharmacokinetic obtained parameters will be maximal plasma concentration, minimal plasma concentration, predose concentration, 24 hour post predose concentration, the time for maximum concentration. The area under the curve will be determined by the trapezoidal rule. Male Versus female data will be compared using 2-tailed t-test. RESULTS/ANTICIPATED RESULTS: We anticipate that: (1) Hispanic/Latino women will have longer circulating times of the drug in the bloodstream and higher maximum concentrations, compared with men. (2) Hispanic/Latino men and women will have higher amounts of the circulating drug, compared with already reported pharmacokinetic data of non-Hispanic Caucasian men. DISCUSSION/SIGNIFICANCE OF IMPACT: Gender differences have been elucidated in the prevalence rates of substance abuse, health service utilization, treatment outcomes, and physiological consequences of drug consumption in the United States. It is known that in general, women progress from drug use to dependence must faster than men; women also suffer more severe physical and emotional consequences than men, yet women seek treatment for drug addiction in lower rates compared with men. Women also show lower pharmacological treatment effectiveness as they are less likely to feel satisfied upon entering a substance abuse treatment and they show higher cravings. Sublingual buprenorphine is a very popular and relatively new medication used primarily for opiate addiction since 2002. Gender differences have been elucidated in the pharmacology of buprenorphine sublingual tablets used for the treatment of opioid addiction. One study showed that women had higher concentrations of circulating parent drug and it is metabolites compared with men. One metabolite in particular norbuprenorphine was found in almost double the plasma concentration in women. Interestingly, gender differences were not pursued at all by the Pharmaceutical Company sponsoring the approval of the sublingual Buprenorphine by the FDA. The cytochrome enzyme CYP 3A4 responsible for the metabolism of Buprenorphine has higher activity in Caucasian/African American women compared with men. However these studies failed to design and recruit significant amount of patients with Hispanic ethnicity to adequately elucidate the gender differences within this ethnic group. Higher plasma concentrations and longer circulation times of a drug may result not only in lower efficacy outcomes but also higher toxicity and undesired effects. Unfortunately, the lack of pharmacological effectiveness and lack of satisfaction in women undergoing drug treatment programs has not been adequately studied to understand the gender difference in pharmacological treatment outcomes between Hispanic/Latino men and women. Due to the under-representation of Hispanic/Latino men but most importantly women in s studying the pharmacology of sublingual Buprenorphine, and considering the well-established gender difference of the principal enzyme (CYP 3A4) responsible for the pharmacology of Buprenorphine, we are proposing a pilot study of the pharmacology of sublingual Buprenorphine in Hispanic/Latino volunteers living in Puerto Rico with equal number of male and female patients. We expect our research to clinically and scientifically elucidate the gender differences of sublingual buprenorphine for opioid addiction in Hispanics/Latinos. The outcome of such research will be the foundation of subsequent clinical studies that aim in updating the current standard of care for Hispanic/Latino men and women that require therapy for opioid addiction.

OBJECTIVES/GOALS: Pursuit of independent funding by predoctoral and postdoctoral fellows requires navigating the intricate steps in preparing extramural grant applications. The Workforce Development Core of the NJ Alliance for Clinical and Translational Science (NJ ACTS) sought to evaluate an interactive grant writing group of fellows mentored by a trained coach. METHODS/STUDY POPULATION: Participants meet weekly for 3 months to develop components of a fellowship application for submission to NIH and private foundations. Sessions were moderated by a senior faculty member trained as a coach by the National Research Mentoring Network. Participant grant submission and review of the program were collected annually for the period of 2019 to 2021 as well as demographics for the 2022 cohort. RESULTS/ANTICIPATED RESULTS: Over this period, 32 predoctoral and 19 postdoctoral fellows participated in the peer review writing groups with 24 trainees currently enrolled. The peer review writing group moved to Zoom in 2020 which has enabled expansion of training to include the 3 Hub institutions and 6 additional universities. Of the 41 survey respondents, 78% submitted fellowship applications to NIH (N=28) or a non-NIH agency (N=4). Eight of these applications are currently under review or have been resubmitted for peer review. 54% of reviewed applications have been funded as NIH fellowships, diversity supplements, career grants, or non-NIH fellowships. Over 90% of participants have recommended the writing group to other trainees. DISCUSSION/SIGNIFICANCE: In conclusion, a weekly grant writing group of predoctoral and postdoctoral fellows is an effective means to receive peer review of fellowship application components and support submissions for extramural funding.

OBJECTIVES/GOALS: Clinical research degree program accreditation brings value to university programs situated in competitive environments. While the requirements of accreditation can be burdensome and tedious, it remains important to program growth. The objective of this project was to assess the use of REDCap for the accreditation process to reduce that burden. METHODS/STUDY POPULATION: A review of credentialing requirements was conducted to identify required data and its sources. Initial course data from a small sample of courses was collected in Excel to better assess the order of the data collection process. REDCap was then used to create a series of data collection instruments that effectively met the program evaluation data needs and customized reports for three years of course learning outcomes. The instruments were developed for its translation to other programs. Faculty, administrators, and interns participated in 2 different types of data collection activities (excel and REDCap) and evaluated the differences between the experiences. RESULTS/ANTICIPATED RESULTS: Data collection included 85 courses, with a range of 3-22 objectives that classified aligned assignments among 8 clinical research professional domains, 50 competencies, and 3 learning levels. Student outcomes data was also calculated and recorded. The time to complete the data collection process using the REDCap tool verses the excel spreadsheet per course was notably more efficient. User satisfaction was 100% improved using the REDCap tool with the average score of 8.5 out of a 1-10 scale. User comments supporting the REDCap process focused on improved time to complete and ease of process. DISCUSSION/SIGNIFICANCE: The incorporation of REDCap into data collection for program accreditation data requirements highlights the efficiency and ease of electronic data capture compared to manual entry in excel. The development of instruments makes it easy to translate to other program evaluation and accreditation needs.

OBJECTIVES/GOALS: The objective of this project was to evaluate the factors that contribute to LGBTQIA2+ patient comfortability. This information was then used to understand how best to create a comfortable space for LGBTQIA2+ patients. METHODS/STUDY POPULATION: This survey was focused on underinsured and uninsured patients seen at the Rainbow Clinic - a free student-run LGBTQIA2+ clinic. Surveys were distributed by undergraduate volunteers on tablets as a qualtrics survey. Surveys collected demographic information in addition to 5 questions that assessed patient comfortability. These questions included evaluating the patient’s comfort with sharing information with the provider and the patient’s comfort of coming into clinical spaces. These surveys were distributed before and after clinic appointments to capture any changes in comfortability that could have occurred as a result of the appointment. RESULTS/ANTICIPATED RESULTS: Up to May of 2023, 49 patients were seen in Rainbow Clinic. 33 patients filled out the intake survey and 31 patients filled out the check-out survey resulting in a 67% and 63% response rate respectively. Questions were asked on a likert scale (1-5) from Strongly Disagree to Strongly Agree. Questions evaluating patient comfort in sharing information with their provider yielded an average score that was statistically significant, suggesting patients felt comfortable at the Rainbow Clinic. Additionally, patients indicated that the LGBTQIA2+ specific labeling of the Rainbow Clinic made them significantly more comfortable coming into the clinic. DISCUSSION/SIGNIFICANCE: This project suggests that patient comfortability can be improved by training and intentional LGBTQIA2+ labeling. Considering the hesitancy of this community towards healthcare, improving comfortability not only benefits clinical care and outcomes but can also bolster the body of research on this community.

OBJECTIVES/GOALS: Atopic dermatitis is one of the most common chronic childhood conditions worldwide and is associated with poor mental health outcomes. Our aim is to determine whether childhood atopic dermatitis is associated with symptoms of depression throughout childhood and adolescence, and whether this association is mediated by serum inflammatory markers. METHODS/STUDY POPULATION: We will perform a longitudinal analysis of over 7000 children from an existing prospective cohort. The primary exposure is atopic dermatitis (AD) annual period prevalence measured by a standardized questionnaire at 12 time points between age 6 months and 16 years. Depression is measured using self-reported responses to the Short Moods and Feelings Questionnaire at 6 time points between 10 and 18 years of age. Cross-sectional regression analyses will be performed to compare depressive signs between children with and without AD and test for dose-response effects with AD and depression. Longitudinal analyses will be conducted using mixed-effects models to estimate the average effect across childhood. We will complete a mediation analysis to determine the extent to which IL-6 and CRP mediate this association. RESULTS/ANTICIPATED RESULTS: We anticipate that atopic dermatitis will be associated with SMFQ scores in a dose response relationship, and that inflammatory markers CRP and IL-6 will partly mediate this association. DISCUSSION/SIGNIFICANCE OF IMPACT: Childhood is a critical time for mental health. Understanding the longitudinal relationship between atopic dermatitis, depression, and inflammatory mediators is crucial as new biologic treatments targeting inflammatory cascades are approved for atopic dermatitis and have the potential to prevent mental health conditions.

OBJECTIVES/SPECIFIC AIMS: To date, only 1 documented case of an individual cured of HIV has been reported. He received an allogeneic bone marrow transplant with cells harboring an HIV-resistant genotype. To mimic this result, we have initiated a Phase I to evaluate the safety of an autologous stem cell gene therapy bone marrow transplant in HIV-related lymphoma patients. METHODS/STUDY POPULATION: The first cohort of patients will receive a 1:1 ratio of unmanipulated CD34 hematopoietic stem cells (HSC) and lentivector modified CD34 HSC expressing a combination of HIV-resistant genes and a selectable marker for cell sorting prior to transplantation. Safety of the HIV-resistant stem cells will be assessed by evaluating engraftment, expression of the anti-HIV genes, and the stability and sequence of the vector. RESULTS/ANTICIPATED RESULTS: One patient has been enrolled and transplanted with the HIV-resistant stem cells. After 1 and 2 months post-transplant, patient blood samples were received, processed for genomic DNA, analyzed by quantitative PCR (qPCR), and displayed successful engraftment of 16 and 12 vector copies per 100 cells, respectively. Expression of all anti-HIV genes was confirmed by qPCR. PCR on genomic DNA confirmed the correct sizes and sequences of the integrated vector and confirmed the successful engraftment of our gene modified cells. Currently, we are enrolling more patients into the trial. DISCUSSION/SIGNIFICANCE OF IMPACT: If successful, this therapy has the potential to change HIV treatment.

OBJECTIVES/GOALS: Compiling information about characteristics and progress of scholars is required for career development applications and progress reports. The range of information is substantial, and preparation is onerous. We sought to create a tool to facilitate gathering key data about trainees and mentors who participate in programs like NIH K- and T-awards. METHODS/STUDY POPULATION: Using the REDCap platform, we developed forms and surveys to support intake of applicants, updates from scholars, and information about their participation in activities and use of resources. We deployed application programming interfaces (APIs) to automate capture of publicly available data about publications, impact metrics, and federal grant funding. Similar tools capture descriptions of mentor expertise including grant funding, prior trainees, and publications with mentees We also built modules to 1) allow connection to institutional grant and contract data to capture foundation and other funding; 2) pre-populate follow-up surveys to update information about career trajectories with minimal scholar effort; and 3) support mentee-mentor agreements as living documents. RESULTS/ANTICIPATED RESULTS: After a pilot period at our institution, we disseminated Flight Tracker to more than 50 academic institutions, most of whom are CTSA hubs. They track scholars in TL1/T32s, KL2/K12s, MSTP programs, and academic groups. Beyond federal reporting, uses now include publication impact factors (relative citation ratios, Altmetrics scores), grant funding of groups, maps of network relationships among investigators, scholar receipt of internal pilot awards, and statistics about transition to independence and time-to-promotion. Scholars can be separated into smaller cohorts by demographics, training dates, and funding dates. Over 34,000 scholars are tracked nationally among over 260 programs. Having structured data supports program evaluation, continuous improvements, and documents program strengths. DISCUSSION/SIGNIFICANCE: We informally estimate Flight Tracker reduces staff and leadership effort in preparation of program data by 75%, preserving time to focus on service to scholars. Ready access to data over time and within and across institutions creates new opportunities for collaborative data analysis to support evidence-based career development.

OBJECTIVES/GOALS: A workflow management tool is essential in order to help support consistent processes with transparency in next steps of the study process. Prior to this tool, staff has relied upon extensive training and coaching on the study process. While resources and guidelines exist, it requires additional time for staff to identify these resources and allows for confusion and rework. Implementation of a systematic workflow management tool was identified as a critical need in order to support streamlined processes, improve transparency and support business continuity, and to accelerate the study process. METHODS/STUDY POPULATION: This effort was undertaken as part of the Protocol Lifecycle Management effort to implement a comprehensive clinical trial management system for clinical research studies. Mayo Clinic has designed a workflow management tool within the Velos eResearch system. The workflow manager is dynamic and will present specific activities based on the study design and responses to data entered on the ad hoc forms. A Workflow Build group contributed to the design of the workflow in order to reflect appropriate, current operational processes. The workflow was vetted and validated with research teams. In addition to designing activities, planned dates and target timelines were established for relevant workflows to help promote transparency in the study start-up timelines and allow study staff to identify overdue activities. Study status controls were designed in the workflow to protect study staff from inadvertently changing the status until appropriate activities are complete. RESULTS/ANTICIPATED RESULTS: A dynamic workflow has been designed and implemented in the Velos eResearch system to support Mayo Clinic research sites. This system will be implemented February 24, 2020 to all consenting studies. DISCUSSION/SIGNIFICANCE OF IMPACT: The implementation of this workflow management tool is critical to help support research operations in a large, academic medical center. Benefits to implementation are expected to include improved transparency in the study status and next steps, reductions in rework due to confusion in next steps, better understanding from new staff in the appropriate study process, and improved timelines for study start-up. As we prepare for the implementation of the Velos eResearch system at Mayo Clinic, the workflow management tool has been identified in training sessions as a positive benefit.

OBJECTIVES/SPECIFIC AIMS: Patients with locally advanced pancreatic cancer typically have poor outcomes, with a median survival of ~16 months. Novel methods to improve local control are needed. Nab-paclitaxel (abraxane) has shown efficacy in pancreatic cancer and is FDA approved for metastatic disease in combination with gemcitabine. Nab-paclitaxel is also a promising radiosensitizer based on laboratory studies, but it has never been clinically tested with definitive radiotherapy for locally advanced disease. METHODS/STUDY POPULATION: We performed a phase 1 study using a 3+3 dose-escalation strategy to determine the safety and tolerability of dose escalated nab-paclitaxel with fractionated radiotherapy for patients with unresectable or borderline resectable pancreatic cancer. Following induction chemotherapy with 2 cycles of nab-paclitaxel and gemcitabine, patients were treated with weekly nab-paclitaxel and daily radiotherapy to a dose of 52.5 Gy in 25 fractions. Final dose-limiting toxicity (DLT) determination was performed at day 65 after the start of radiotherapy. RESULTS/ANTICIPATED RESULTS: Nine patients received nab-paclitaxel at a dose level of either 100 mg/m2 (n=3) or 125 mg/m2 (n=6). One DLT (grade 3 neuropathy) was observed in a patient who received 125 mg/m2 of nab-paclitaxel. Other grade 3 toxicities included fatigue (11%), anemia (11%), and neutropenia (11%). No grade 4 toxicities were observed. With a median follow-up of 8 months (range 5–28 months), median survival was 19 months and median progression-free survival was 10 months. Following chemoradiation, 3 patients underwent surgical resection, all with negative margins and limited tumor viability. Of the 3 patients, 2 initially had borderline resectable tumors and 1 had an unresectable tumor. Tumor (SMAD-4, Caveolin-1) and peripheral (circulating tumor cells and microvesicles) biomarkers were collected and are being analyzed. DISCUSSION/SIGNIFICANCE OF IMPACT: The combination of fractionated radiation and weekly nab-paclitaxel was safe and well tolerated. This regimen represents a potentially promising therapy for patients with unresectable and borderline resectable pancreatic cancer and warrants further investigation.

OBJECTIVES/GOALS: Academic research centers often struggle to recruit and retain a diverse and competent clinical and translational science (CTS) workforce. Specifically, the clinical research professional (CRP) career pathway is not well known to undergraduate students and other individuals outside of academic medicine despite various potential career routes. METHODS/STUDY POPULATION: To address these workforce challenges, the CRP Task Force at the University of Cincinnati (UC) aims to train a competent and diverse CRP workforce through targeted educational programming in the UC undergraduate population. Using a six-step curriculum development process that included: 1) performing a needs assessment, 2) determining content, 3) writing goals and objectives, 4) selecting the educational strategies, 5) implementing the curriculum, and 6) evaluating the curriculum, we designed an undergraduate certificate program in CTS. RESULTS/ANTICIPATED RESULTS: The needs assessment included both internal and external data gathering to inform curriculum development and program decisions. Content was determined using the CRP Competency Framework 2.0, and program learning outcomes were written with both the competency framework and local workforce needs in mind. Educational strategies were selected based upon optimization of available resources and local expertise with an emphasis on interactive didactics complemented by experiential learning. Implementation is underway and evaluation will follow once students begin enrolling. DISCUSSION/SIGNIFICANCE: We anticipate an increase in numbers of well-qualified, diverse applicants who pursue CRP careers locally and regionally. In addition, we expect that the certificate program will build competency earlier in CRP staff, improving job satisfaction and retention as a result of a stronger foundation from which to build their professional skills.

OBJECTIVES/SPECIFIC AIMS: Lead (Pb) exposure can seriously affect nervous system and kidney. Young children are vulnerable to Pb exposure. However, the role of low-level Pb exposure in asthma in children and the age and racial disparity is not well studied. The objectives are to examine the relation between Pb level and asthma status and to determine the age and racial/ethnic differences in this relation. METHODS/STUDY POPULATION: We analyzed data from National Health and Nutrition Examination Survey 1999-2016 for 22,885 children 1-15 years old. Asthma information was collected by questionnaire. Blood lead level was measured using mass spectrometry. The association between blood Pb level and asthma status was assessed by logistic regression after adjusting for children’ age, gender, race/ethnicity, insurance status, and source of care; household poverty, mother’s age and smoking status. Data were analyzed using Stata 14 considering design and sample weight and p<0.05 is statistically significant. RESULTS/ANTICIPATED RESULTS: Pb level was associated with asthma status (Adjusted Odds Ratio (AOR)=1.4, 95% Confidence Interval (CI) = 1.2-1.7, p < 0.001). Stratified analysis by age showed that blood Pb level is related to asthma only in children 1-5 years old (AOR = 1.3, 95% CI = 1.1-1.5, p = 0.004). There was no racial/ethnic difference in this association. DISCUSSION/SIGNIFICANCE OF IMPACT: Pb level is associated with asthma status in children especially young children. Health risk of low Pb is a concern. Preventive measures by reducing potential sources of Pb should be introduced early.

OBJECTIVES/GOALS: Murine and clinical data suggest that antibody-dependent cellular cytotoxicity (ADCC) is associated with greater protection against disseminated neonatal HSV disease. To quantify the relative transfer of Abs with different functions and targets, we conducted a prospective study of mother-infant term and preterm dyads pre and during COVID-19 METHODS/STUDY POPULATION: Total and HSV lysate, glycoprotein D (gD) and glycoprotein B (gB)-specific IgG, IgG1 and IgG3 as well as HSV neutralizing Abs (nAbs) and ADCC were quantified in paired 3rd-trimester pregnant women and their newborns (cord) blood. Transfer ratios (TR) were defined as cord:maternal Ab levels. IgG1 and IgG3 subclass and gD or gB-specific Abs were isolated by column purification and glycan profiles were assessed by mass spectrometry. The study population included 21 term and 15 preterm dyads who were HSV-1 (+/- HSV-2) seropositive enrolled between 2018-2019 (pre-COVID) and 25 additional HSV-1 (+/- HSV-2) seropositive term dyads whose mothers were SARS-CoV-2 PCR and COVID Ab+ at delivery; 14 were asymptomatic and 11 had mild-moderate COVID disease. None of the mothers had active genital HSV lesions during delivery RESULTS/ANTICIPATED RESULTS: Anti-HSV IgG, IgG1 and IgG3 TR were higher in term vs. preterm dyads (p<0.05). The nAb TR was 2.4 in term vs. 0.8 in preterm (p<0.001) but the ADCC TR was < 1.0 for both. To determine if the latter reflected antigenic target, subclass or glycans, we enriched for gD and gB specific and IgG1 and IgG3 Abs. The gD Abs were IgG1 and had only neutralizing activity. In contrast, gB Abs were polyfunctional and included IgG1 and IgG3 but only the IgG1 Abs had ADCC activity. The gD Abs were enriched for glycans associated with an affinity for the neonatal Fc receptor (FcRn); gB Abs expressed glycans associated with both FcRn and FcγRIIIa binding. There was no significant difference in total HSV-specific IgG TR in pre-COVID vs post-COVID dyads but the nAb TR was lower (p=0.018) and ADCC TR higher (p<0.001) in the COVID compared to pre-COVID cohort DISCUSSION/SIGNIFICANCE: HSV ADCC Abs, which may provide greater protection than nAbs against neonatal disease, transfer poorly particularly to preterm newborns. However, in the setting of SARS-CoV-2, the TR of HSV ADCC is significantly higher. This may reflect alterations in the placental architecture and/or glycan composition which is currently being investigated.

OBJECTIVES/GOALS: Young and middle-aged adults with a myocardial infarction (MI) represent an understudied group potentially with unique risk indicators such as emotional stress. We sought to investigate if mental stress-induced myocardial ischemia (MSIMI), a marker of cardiovascular vulnerability to psychological stress, is associated with poor outcomes among this population. METHODS/STUDY POPULATION: We studied 306 patients (150 women and 156 men) ≤61 years of age who were hospitalized for MI in the previous 8 months. Clinical, behavioral and psychosocial factors were assessed with standardized measures. Patients underwent myocardial perfusion imaging with mental stress (public speaking) and conventional stress (exercise or pharmacological testing). MSIMI and conventional stress-induced ischemia were defined as a new or worsening perfusion defect. Patients were followed for 3 years for adverse events, which were independently adjudicated. Cox proportional hazard models were used to estimate the association of MSIMI and CSIMI with a composite endpoint of recurrent MI or cardiovascular (CV) death with adjustment for demographic, clinical and psychosocial risk factors. RESULTS/ANTICIPATED RESULTS: The mean age of the sample was 50 years (range, 22-61). MSIMI occurred in 16% of the patients, and conventional ischemia in 35%. Over a 3-year follow-up, 28 individuals had a recurrent MI and 2 died due to cardiovascular causes. The incidence of the composite endpoint of MI or CV death was more than doubled in patients with MSIMI (20%) than those without MSIMI (8%), HR 2.6, 95%CI, 1.2-5.6. Further adjustment for demographic and clinical risk factors and depressive symptoms did not substantially change the relationship. In contrast, conventional stress ischemia was not significantly related to the outcome (HR 1.4, 95%CI, 0.6-3.0). DISCUSSION/SIGNIFICANCE OF IMPACT: Young and middle-aged individuals with MSIMI after MI have a >2-fold higher likelihood of recurrent MI and CV mortality compared with those without MSIMI. In this patient group, MSIMI is a better risk indicator than ischemia with a conventional stress. These findings point to psychological stress as an important determinant of risk in this patient population. Ischemia induced by mental stress is a potent risk indicator in young post-MI patients. Stress-reduction interventions may be especially beneficial in patients who show this abnormal response.

OBJECTIVES/SPECIFIC AIMS: Systemic sclerosis (SSc) is a connective tissue disease of unknown etiology characterized by progressive fibrosis of the skin and multiple visceral organs. Effective therapies for SSc are needed. Lysyl oxidase (LOX) is a copper-dependent amide oxidase that plays a critical role in the crosslinking of the extracellular matrix (ECM). In this study, we investigated the role of LOX in the pathophysiology of SSc. METHODS/STUDY POPULATION: LOX expression and protein levels were measured in lung tissues and primary fibroblasts from patients with SSc and healthy controls. The effects of recombinant LOX (rLOX) were measured in vitro in primary fibroblasts, ex vivo in human lung tissues and in vivo in mice given bleomycin in combination with rLOX. LOX levels and activity were evaluated in lung fibroblasts treated with an endostatin-derived peptide that ameliorates fibrosis and in mice treated with bleomycin in combination with the peptide. Further, to differentiate the crosslinking activity of LOX from other potential effects, primary human fibroblasts were cultured with rLOX in the presence of the inhibitor, beta-aminopropionitrile. The expression levels of ECM (collagen and fibronectin), pro-fibrotic factors (IL-6 and TGF-beta), and transcription factor (c-Fos) were examined by real-time PCR, ELISA, immunoblotting, or hydroxyproline assay. RESULTS/ANTICIPATED RESULTS: LOX mRNA was increased in lung tissues and matching fibroblasts of SSc patients. rLOX-induced ECM production in vitro and ex vivo in lung fibroblasts and in human lung tissues maintained in organ culture, respectively. Additionally, TGF-beta and bleomycin induced ECM production, LOX mRNA expression and activity. Endostatin peptide abrogated these effects. In vivo, rLOX synergistically exacerbated pulmonary fibrosis in bleomycin-treated mice. The inhibition of LOX catalytic activity by beta-aminopropionitrile failed to abrogate LOX-induced ECM production. LOX increased the production of IL-6. IL-6 neutralization blocked the effects of LOX. Further, LOX induced c-Fos expression and its nuclear localization. DISCUSSION/SIGNIFICANCE OF IMPACT: LOX expression and activity were increased with fibrosis in vitro, ex vivo, and in vivo. LOX induced fibrosis via increasing ECM, IL-6 and c-Fos translocation to the nucleus. These effects were independent of the crosslinking activity of LOX and mediated by IL-6. Our findings suggest that inhibition of LOX may be a viable option for the treatment of lung fibrosis. Further, the use of human lung in organ culture establishes the relevance of our findings to human disease.

Objectives/Goals: Infectious keratitis is the leading cause of corneal blindness worldwide, causing two million cases of monocular blindness per year. Of these cases, developing countries are disproportionately affected, in part due to sociodemographic disparities. Our study examined risk factors for severe keratitis presentation in a South Indian population. Methods/Study Population: 156 patients aged ≥ 16 years with clinically diagnosed infectious keratitis presenting to Aravind Eye Hospital in Pondicherry, India, from January 1, 2023 to July 31, 2024, were retrospectively reviewed. Univariate logistic regression was used to evaluate associations between specific potential risk factors (including age, sex, awareness of keratitis, travel distance to hospital, education level, missed work wages, and ability to afford care) and keratitis clinical severity (defined using thresholds of poor visual acuity, size of stromal infiltrate measured on slit lamp examination, occurrence of perforation, and need for corneal transplant surgery). Individual risk factors found to be significant were incorporated into a multivariable logistic regression model. Results/Anticipated Results: We anticipate that the results of this study will identify multiple risk factors for more severe keratitis presentations among patients at baseline. We expect these factors to include increased travel distance from the patient’s home to the base hospital, delays between time of diagnosis and initiation of treatment, treatment nonadherence, lower educational levels, lack of familiarity with keratitis, treatment and transportation costs, increased time off work, and missed work wages, among others. Discussion/Significance of Impact: This study elucidates barriers to early keratitis diagnosis in a low-resource setting. Study findings can inform strategies to improve keratitis prevention using decentralized care approaches such as community eye screenings and expanding outreach via vision centers. Such strategies can improve timely access to care for vulnerable populations.