OBJECTIVES/GOALS:

1. 1. Determine test characteristics of Neuro ICU nurse interpretation of real-time bedside qEEG for seizure detection

2. 2. Determine difference in time to detection of seizures between qEEG interpretation and raw cEEG reads

3. 3. Determine how seizure characteristics affect accuracy of qEEG reads

METHODS/STUDY POPULATION:

• Subjects: Nurses caring for patients admitted to the Neuro ICU at Duke University Hospital who are initiated on cEEG.

• Nurses evaluate qEEG display at the bedside on an hourly basis after undergoing a standardized qEEG training session. The standard practice of independent review of cEEG and treatment by the Neuro ICU team remains unchanged.

• Post-hoc review of cEEG data by two blinded, board-certified neurophysiologists will be performed for each patient. The raw cEEG data will be scored for the number of seizures present per hour, background, seizure duration, and seizure spatial extent.

• The time from first seizure occurrence to clinical recognition will be recorded.

RESULTS/ANTICIPATED RESULTS:

• Thus far, 91 patients with 583 1-hour blocks of nurse interpretations have been studied, with 6 patients experiencing seizures while on study. Enrollment will be completed on 1/17/20

• Preliminary data show a sensitivity of 95.8% (79.9%, 99.9%), specificity of 95.2 (93.1%, 96.8%), positive predictive value of 46.0% (36.9%, 55.4%), negative predictive value of 99.8% (98.7%, 99.9%), positive likelihood ratio of 19.8 (13.6, 28.9), negative likelihood ratio (0.04 (0.01, 0.3). All confidence intervals are 95%. False alarm rate is 0.05/hour.

• Further analyses are pending completion of enrollment in January 2020.

DISCUSSION/SIGNIFICANCE OF IMPACT: Nurse interpretation of real-time bedside qEEG for seizure detection is feasible in the Duke Neuro ICU. QEEG functions well as a screening tool with good specificity and low false alarm rate. Use of qEEG by nurses could lead to shorter time to seizure detection, which may improve patient outcomes. CONFLICT OF INTEREST DESCRIPTION: Safa Kaleem, BS: Research reported in this publication was supported by a Pfizer Foundation grant and the Duke Clinical Translational Science Institute (CTSI). The content is solely the responsibility of the authors and does not necessarily represent the official views of the Pfizer Foundation or the Duke CTSI. Jennifer H. Kang, MD: None to declare. Alok Sahgal, MD: None to declare. Christa B. Swisher, MD: Received speaker’s honorarium from EISAI and UCB.

OBJECTIVES/GOALS: Diabetes is related to risk for heart disease, stroke, high blood pressure, and COVID-19. It is exacerbated by built/social environment issues, e.g., food insecurity, access to healthy foods and health care, and other poverty-related factors. Our goal is to assess the efficacy of an integrated care model for patients with poorly controlled diabetes. METHODS/STUDY POPULATION: We utilize an integrated, team-based approach to diabetes treatment. In a traditional care model, too little focus is on social determinants and their impacts on health and well-being. Our project involves enrollment of patients with diabetes in an intervention whereby their medical care is integrated with intensive diabetes education and provision of social and other health services, including diet and nutrition, exercise, provision of foods and nutritional supplements, and other support services as needed to achieve optimal health and to reduce morbidity and unnecessary hospitalization and emergency room visits. Subjects are underserved patients treated through our non-profit community clinic partners. We track metrics including individual outcomes, organizational outcomes, and collective impact. RESULTS/ANTICIPATED RESULTS: We anticipate that patients enrolled in this study will demonstrate significant improvements in diabetes control and management. Clinical improvements will include better glycemic control, improved hypertension and dyslipidemia management, reduced complications, and increased preventive measures including foot, eye and oral health exams and monitoring of microalbuminuria. Overall, we anticipate decreased frequency of hospitalizations and readmissions as well as decreased frequency of emergency care visits for treatment of diabetes-related issues. We expect patients to experience enhanced self-efficacy, increased physical activity, and improved quality of life. Their outcomes will be compared to controls receiving the standard medical regimen, matched on age, race, gender, and time of onset. DISCUSSION/SIGNIFICANCE: These activities will improve understanding of factors influencing diabetes outcomes at individual and upstream levels. It will inform food distribution and models of care for improved patient outcomes, including social determinants of health and will establish new protocols for community-based provision of health care to our most vulnerable.

Objectives/Goals: Urinary tract infections (UTIs) cause significant morbidity, and many patients require multiple courses of antibiotics increasing the risk of antibiotic resistance. We determined the prevalence of urinary antibiotic heteroresistance (HR), which has been associated with treatment failures in vivo, to three first-line antibiotics for UTIs. Methods/Study Population: Clinical urine Escherichia coli isolates from patients in metropolitan Atlanta, Georgia in August 2023 were collected as part of public health surveillance performed by the CDC-funded, Georgia Emerging Infections Program (EIP). Only the first E. coli isolate collected for each patient was included in this study. Antibiotic susceptibility was determined through medical record review. HR to nitrofurantoin, trimethoprim-sulfamethoxazole, and fosfomycin was determined by population analysis profiling (PAP), where broth dilutions of E. coli were plated on increasing concentrations of the antibiotic. HR was defined as survival of >1 in 106 cfu but fewer than 50% survival at 1X antibiotic breakpoint (bp), resistant as > 50% survival at 1X bp and susceptible as survival of Results/Anticipated Results: Among 355 patients, 21 (5.9%) were resistant or intermediate to nitrofurantoin and 92 (26%) were resistant to trimethoprim-sulfamethoxazole. Antibiotic susceptibility data were missing from 5(1.4%) and 11(3%) of isolates for nitrofurantoin and trimethoprim-sulfamethoxazole, respectively. Susceptibility testing was not routinely performed nor reported for fosfomycin, thus excluded. PAP revealed that of the total 355 isolates, 3(0.84%) were heteroresistant to nitrofurantoin, 17(4.8%) were heteroresistant to trimethoprim-sulfamethoxazole, and 27(7.6%) were heteroresistant to fosfomycin. Of the isolates found to be susceptible by standard testing, 1(0.3%) and 9(3.6%) were heteroresistant to nitrofurantoin and trimethoprim-sulfamethoxazole by PAP, respectively. Discussion/Significance of Impact: Despite low rates of HR to nitrofurantoin and trimethoprim-sulfamethoxazole (0.84%, 4.8%), HR to fosfomycin was more frequent (7.6%). Given that susceptibility is not generally performed for fosfomycin, this could have implications for including fosfomycin as a first-line treatment for E. coli UTIs.

OBJECTIVES/GOALS: Despite a steady rise of graduate degrees in biostatistics earned in the US, the percent from minorities remains low. This poster will describe the Collaborative Undergraduate Biostatistics Experience (CUBE), an 8-week program aimed to diversify and bring awareness to the field of collaborative biostatistics, from recruitment through evaluation. METHODS/STUDY POPULATION: The CUBE program is funded jointly by the NIH’s NIDA/NIAAA (award number: 1R25DA058482-01) and is designed to give underrepresented minority (URM) undergraduate students in STEM the opportunity to engage in a collaborative biostatistics and health data science experience, along with related professional development activities. The program is built on four pillars: 1) training in introductory biostatistics, 2) training in R programming, 3) professional development, and 4) a collaborative research project addressing research questions in various disciplines. The CUBE program was delivered in the summer of 2022 as a pilot to four URM students at Virginia Tech (VT) and the University of Virginia (UVA), with two at each site. In summer 2023, the program was offered to 5 students (3 VT, 2 UVA). RESULTS/ANTICIPATED RESULTS: This poster will provide strategies learned over two summers with respect to recruitment and enrollment, along with details on the program content, timeline, and short- and long-term program evaluation metrics (both quantitative and qualitative). The CUBE program was well-received by students participating in summers 2022 and 2023, where improved attitudes towards statistics were demonstrated, and 7 of the total 9 participants (78%) over the past two summers expressed interest in pursuing a graduate degree in biostatistics or a career in quantitative research. Of these 7 students, 1 is currently enrolled in a biostatistics graduate program in the United States. DISCUSSION/SIGNIFICANCE: Results can be used to offer recommendations to leaders in the field on how to establish similar programs seeking to provide a pipeline for equity and diversity in the practice of collaborative biostatistics and health data science.

OBJECTIVES/SPECIFIC AIMS: Patients are increasingly using online materials to learn about gynecologic cancer. Recent studies demonstrate that 85-96% of patients with a gynecologic malignancy utilize the Internet as a health resource. Providers can refer patients to educational materials produced by major medical associations available on their websites. However, patient educational materials (PEMs) published by professional organizations from other surgical specialties have been shown to be difficult to read for the average American. The NIH and AMA recommend that PEMs be written between a sixth and eighth grade reading level. In this study, we assess the readability of online PEMs on gynecologic cancer published by major medical associations. METHODS/STUDY POPULATION: Seven national medical association websites with PEMs on gynecologic malignancy were surveyed: American College of Obstetricians and Gynecologists, Center for Disease Control, Foundation for Women’s Cancer, National Cancer Institute, National Cervical Cancer Coalition, National Ovarian Cancer Coalition, and Society of Gynecologic Oncology. Online PEMs were identified and analyzed using five validated readability indices. One-way ANOVA and Tukey’s test were performed to detect differences in readability between publishers. RESULTS/ANTICIPATED RESULTS: Two hundred and thirty PEMs were included in this analysis. Mean readability grade levels with standard deviation were: 11.3 (2.8) for Coleman-Liau index; 11.8 (3.2) for Flesch-Kincaid; 11.1 (1.2) for FORCAST formula; 12.5 (2.7) for Gunning FOG formula; 12.1 (2.6) for New Dale-Chall formula; and 13.5 (2.5) for SMOG formula. Overall, PEMs were written at a mean 12th grade reading level. Only 4.3% of articles were written at an 8th grade reading level or below. ANOVA demonstrated a significant difference in readability between publishing associations (p<0.01). PEMs from the Center for Disease Control had a mean 10th grade reading level and were significantly lower than all other organizations. PEMs from The Foundation for Women’s Cancer had a mean 13th grade reading level and were significantly higher than most other organizations. DISCUSSION/SIGNIFICANCE OF IMPACT: Gynecologic oncology PEMs available from major medical association are written well above the recommended sixth to eight grade reading level. Simplifying PEMs may improve patient understanding of their disease and facilitate physician-patient communication.

ABSTRACT IMPACT: This work seeks to improve the diagnostic accuracy of urinary tract infection among hospitalized older adults and mitigate antibiotic overuse in this population. OBJECTIVES/GOALS: Primary objective: To determine the diagnostic accuracy of serum procalcitonin (PCT) for the diagnosis of symptomatic urinary tract infection (UTI) in hospitalized older adults. Secondary objectives: (1) To develop a predictive model for the diagnosis of UTI; (2) To determine the ability of PCT in discriminating between lower and upper UTI. METHODS/STUDY POPULATION: We performed a prospective observational cohort study of 228 participants from a single institution. The study population included older adults (age 65 or older) who were hospitalized on the general medicine wards with a possible or suspected urinary tract infection (UTI). Upon obtaining informed consent, serum procalcitonin (PCT) was processed on remnant blood samples collected from the emergency department. We performed additional data collection through the electronic health record to obtain demographic information, clinical characteristics, and other laboratory and imaging results. Clinicians were surveyed for the diagnosis of UTI and charts were adjudicated by independent reviews of the medical record by infectious diseases experts to determine the primary endpoint of symptomatic UTI. RESULTS/ANTICIPATED RESULTS: We anticipate that serum procalcitonin predicts the presence of symptomatic urinary tract infection (UTI) by demonstrating an area under the receiver operating characteristic curve of at least 0.85. A predictive model developed in our cohort for the diagnosis of symptomatic UTI will be improved by the addition of serum PCT to the prediction model. Finally, we anticipate the serum PCT will accurately discriminate between upper and lower UTI. DISCUSSION/SIGNIFICANCE OF FINDINGS: Diagnosis of symptomatic UTI in hospitalized older adults is challenging and may lead to overuse of antibiotics and the development of antibiotic resistance in this vulnerable patient population. Serum procalcitonin offers a novel diagnostic strategy in the diagnosis of symptomatic UTI to enable more appropriate antibiotic therapy.

OBJECTIVES/SPECIFIC AIMS: To compare the accuracy and precision for estimating the diagnostic accuracies (sensitivities and specificities) between differential verification (DV) and partial verification (PV) methods. Comparisons were made under scenarios with different values of design parameters including disease prevalence, proportion of verification for positive results, proportion of verification for negative result, sensitivity and specificity of the brass standard (BS) test in DV method. Through comparing 2 different verification methods under different scenarios, we give suggestions that which verification method is optimal under different design settings. METHODS/STUDY POPULATION: For both PV and DV methods, simulation studies were performed using statistical package R, version 3.1.3. We were primarily interested in studying how the unbiasedness and precision for estimation of diagnostic accuracies (sensitivity and specificity) of an index test change with the following design parameters: disease prevalence, proportion of verification for positive test results, the proportion of verification for negative test results, and the sensitivity and specificity of a BS test. We chose different values for each of the above parameters. For each estimation, we allowed values in only 1 parameter to change by fixing the other 2 parameters, so that the effect of each design parameter on the unbiasedness and precision of both sensitivity and specificity can be determined. For the DV method, we also developed an analytical method to estimate the sensitivity and specificity of an index test using a quadratic equation with a unique solution of the specificity and sensitivity. RESULTS/ANTICIPATED RESULTS: For rare disease with prevalence less than 1%, the PV method resulted in a less biased and more precise estimate of sensitivities and specificities of the index test. If the disease prevalence was between 1% and 10%, the DV method using a BS test with moderate or high sensitivity and specificity (sensitivity and specificity >90%) resulted in a less biased and more precise estimate of diagnostic accuracies of the index test. When the disease prevalence was greater than 10%, the PV method was superior when the BS test had sensitivity and specificity <80%, and the DV method was superior when the BS test had both sensitivity and specificity >90%. When the proportion of verification of positive test results was <30% or >70%, the DV method yielded smaller bias for the estimated specificity than the PV method. However, the PV method generated a much smaller mean square error (MSE) for specificity than the DV method when the proportion of verification for positive test results was >50%. Although the disease prevalence was >10% and the proportion of verification of positive test results was <30%, the DV method resulted in a smaller MSE for specificity. DISCUSSION/SIGNIFICANCE OF IMPACT: Disease prevalence and proportions of verification for patients with positive and negative test results influence the accuracy of a new diagnostic test. If a new index test for a very rare disease is evaluated, the PV method should be used for assessing the performance of the index test. When a disease prevalence is >1%, the DV method will result in a less biased and more precise estimate of diagnostic accuracy of an index test, if the BS test itself used in the DV method has large specificity and specificity. One concern of using BS test for the DV method is the clinical cost. Depending on the disease type, the BS tests usually are imperfect, but may be less aggressive and/or less expensive than the gold standard test. Moreover, as all clinical examinations require professional personnel to perform, verification of the index test for relative large proportion of a large cohort of patients could become a burden on human resources. Thus, the future research of the optimal design method for a diagnostic accuracy study should be based on the comprehensive cost-effectiveness analysis.

OBJECTIVES/SPECIFIC AIMS: (1) To delineate the function of theheavy-chain receptor binding domain (HCR), a portion of botulinum neurotoxintype A (BoNT/A) and synaptic vesicle 2 (SV2) signaling pathway, whichprovide a novel multipurpose biologic with potential clinical applications intumor detection/imaging, inhibition of tumor progression, andreduction of bioactive hormone secretion in metastatic neuroendocrine (NE)cancers. (2) To evaluate the expression pattern of SV2 receptors in NE cancerpatient-derived tissues for prediction of patient response for recombinant HCR(rHCR) treatment. (3) To assess the in vivo efficacy and toxicity of rHCR in aNE cancer liver metastasis mouse models and in the NE patient-derived 3DMicroTumor system. (4) To collect preclinical data to design and conduct aclinical trial with NE cancer patients, a major goal toward translating ourdiscoveries into much needed therapies. METHODS/STUDY POPULATION:Recombinant botulinum heavy chain (rHCR) was produced using an IPTG-inducibleexpression vector in E. coli BL21. The rHCR was His-Tagpurified and stored in PBS buffer before usage. Cytotoxicity: H727, TT, and MZcells were plated at a density of 5000 cells/well in 96-well platesand incubated under standard conditions overnight. The next day, cells weretreated with 10, 100, or 500 nmol/L of rHCR and incubated for 72hours. Following incubation, cell viability was assessed by ATP quantificationusing the CellTiter-Glo (Promega) assay. Fresh NE tumors were dissociated andinjected into polydimethylsiloxane bioreactors in a matrigel and collagensuspension for 3D culture experiments. The viability of 3D cultures incubatedwith various doses of rHCR was assessed by measuring the uptake of thenear-infrared dye IR-783 using an IVIS imaging system. Western blot: H727, TT,and MZ cells were seeded in 6-well plates at a density of 3×105cells/well for 24 hours followed by treatment with 100nmol/L for 72 hours. Total cellular proteins were isolated andanalyzed to assess the level of SV2A expression and the effect of rHCR on theexpression levels of NET marker proteins. Immunohistochemistry: Deparaffinizedtissue culture slides were incubated with SV2A primary antibody in 1%BSA and incubated overnight at 4°C. Slides were rinsed twice with TBScontaining 0.025% Triton, followed by 0.3%H2O2 for 15 minutes. Slides were then incubated withHRP-conjugated secondary antibody for 1 hour at room temperature. Detection ofprotein-protein interaction: Precleared cell lysate was incubated withglutathione-agarose beads in the presence of 10 μg of GST-tagged rHCRfor 2 h at 4°C with end-over end mixing. Samples were thencentrifuged at 10,000 g for 2 minutes and the supernatant was analyzed bySDS-PAGE. Preclinical models: To allow rHCR testing on NET patient derived cellsin a very novel 3D surrogates, sterile collected fresh NET tissues will beobtained from the UAB Tissue Procurement, dissociated into a single cellsuspension and injected into a polydimethylsiloxane bioreactors containingextracellular matrix composed of bovine collagen and matrigel. Such 3D cellculture will be maintained in bioreactors with constant supply of media throughthe microchannels and treated with rHCR at concentrations ranging from 10 nM to1 mM. Following histologic confirmation of growth and morphology of NETpatient-derived 3D surrogates we will test the anticancer activity of rHCR inthis system using the standard cytotoxicity assays as well as we will validatethe NET hormone expression using immunohistochemistry assay. To create an animalmodel of NE cancer progression, we will perform intrasplenic injection of NETcell lines. In approximately 4 weeks, the animals should develop NE livermetastases based upon our previous experience. rHCR-iFPs accumulation in thetumor mass: rHCR-iFPs will be injected to the tumor bearing mice after 4 weeksof cells implantation in 1 week interval for total of 4 treatments at theconcentrations of 0.125, 1.25, and 12.5 mg/kg.RESULTS/ANTICIPATED RESULTS: Based on the preliminary data, we expectto detect rHCR-iFPs in NE cancer xenografts and in patient derived 3D explants.Our preliminary data revealed that treating NETcells with rHCR significantlyreduced NE peptide expression in 3 days. Thus, we expect to see the decrease ofNE tumor markers even if the fluorescent detection method is not sensitiveenough to monitor the signal. The reduction of the NET markers can be used as anindicator of the rHCR-iFPs uptake by the tumor mass. If HCR exhibit high bindingaffinity to SV2 receptors in NET models but moderate anticancer efficacy, weplan to use rHCR peptide to conjugate with the nanocarrier for targeted drugdelivery. In this case rHCR peptide can be used as a ligand that specificallybinds to NE cancer cells and delivers anticancer drug. In 3D NET patient derivedexplants we expect significant reduction of NET markers and hormones (serotoninand calcitonin) in NE cancer cells upon long-term rHCR-iFPs treatment. Inaddition, we will perform multiplex protein quantification assay using Luminexto assess the various hormones, cytokines, and growth factors to be repurposedinto a diagnostic and detection reagent, or a drug delivery ligand for targetedtherapies. DISCUSSION/SIGNIFICANCE OF IMPACT: NE cancers are highlymetastatic: NE cancers such as carcinoid, islet cell tumors, and medullarythyroid cancer frequently metastasize to the liver. They are the second mostprevalent GI malignancy. Ninety percent of patients with pancreatic carcinoidtumors and 50% of patients with islet cell tumors develop isolatedhepatic metastases. Patients with untreated, isolated NE liver metastases have<30% 5-year survival. Thus, there is a critical need fornew therapies for NE cancers. Surgery is the only curative therapy available forpatients with NE cancers but most cannot be cured: surgical removal is the mosteffective treatment for NE cancers; however, a very high percentage of patientspresent with metastatic disease. While surgical resection can be potentiallycurative, many patients are not candidates for operative intervention due towidespread metastases or the degree of hepatic involvement by the NE cancers.Moreover, other forms of therapy including chemoembolization, radioembolization,cryoablation, and chemotherapy have had limited efficacy. We hope that our invitro data on rHCR toxicity and specificity to NET, validated in thepre-clinical models will allow the first in-human application of this technologyin clinical studies.

The National Center for Advancing Translational Sciences has called for more comprehensive research with priority populations to reduce disparities and for the development of additional resources to assist researchers in implementing these recommendations. Here we report the development and initial evaluation of five Priority Populations Toolkits, which are resources developed by the University of Illinois Center for Clinical and Translational Science to meet these goals. Three aims guide the content: increasing knowledge, facilitating communication, and improving research design. Materials were curated from scientific literature reviews and Internet searches and revised iteratively. Analytics and user surveys provide information about usage. In 22 months, 387 unique users accessed the toolkits. The top reason for usage was to improve research recruitment. Comprehensive toolkits for working with priority populations show promising potential for increasing knowledge and readiness to work with underrepresented populations. Further toolkit development and evaluation of effectiveness are warranted.

The last 50 years have seen an increasing dependence on academic institutions to develop and commercialize new biomedical innovations, a responsibility for which many universities are ill-equipped. To address this need, we created LEAP, an asset development and gap fund program at Washington University in St. Louis (WUSTL). Beyond awarding funds to promising projects, this program aimed to promote a culture of academic entrepreneurship, and thus improve WUSTL technology transfer, by providing university inventors with individualized consulting and industry expert feedback. The purpose of this work is to document the structure of the LEAP program and evaluate its impact on the WUSTL entrepreneurial ecosystem. Our analysis utilizes program data, participant surveys, and WUSTL technology transfer office records to demonstrate that LEAP consistently attracted new investigators and that the training provided by the program was both impactful and highly valued by participants. We also show that an increase in annual WUSTL start-up formation during the years after LEAP was established and implicate the program in this increase. Taken together, our results illustrate that programs like LEAP could serve as a model for other institutions that seek to support academic entrepreneurship initiatives.

Objectives/Goals: Transmission-blocking vaccines hold promise for malaria elimination by reducing community transmission. But a major challenge that limits the development of efficacious vaccines is the vast parasite’s genetic diversity. This work aims to assess the genetic diversity of the Pfs25 vaccine candidate in complex infections across African countries. Methods/Study Population: We employed next-generation amplicon deep sequencing to identify nonsynonymous single nucleotide polymorphisms (SNPs) in 194 Plasmodium falciparum samples from four endemic African countries: Senegal, Tanzania, Ghana, and Burkina Faso. The individuals aged between 1 and 74 years, but most of them ranged from 1 to 19 years, and all presented symptomatic P. falciparum infection. The genome amplicon sequencing was analyzed using Geneious software and P. falciparum 3D7 as a reference. The SPNs were called with a minimum coverage of 500bp, and for this work, we used a very sensitive threshold of 1% variant frequency to determine the frequency of SNPs. The identified SNPs were threaded to the crystal structure of the Pfs25 protein, which allowed us to predict the impact of the novel SNP in the protein or antibody binding. Results/Anticipated Results: We identified 26 SNPs including 24 novel variants, and assessed their population prevalence and variant frequency in complex infections. Notably, five variants were detected in multiple samples (L63V, V143I, S39G, L63P, and E59G), while the remaining 21 were rare variants found in individual samples. Analysis of country-specific prevalence showed varying proportions of mutant alleles, with Ghana exhibiting the highest prevalence (44.6%), followed by Tanzania (12%), Senegal (11.8%), and Burkina Faso (2.7%). Moreover, we categorized SNPs based on their frequency, identifying dominant variants (>25%), and rare variants (Discussion/Significance of Impact: We identified additional SNPs in the Pfs25 gene beyond those previously reported. However, the majority of these newly discovered display low variant frequency and population prevalence. Further research exploring the functional implications of these variations will be important to elucidate their role in malaria transmission.

OBJECTIVES/GOALS: Our primary objective was to determine the demographic and linguistic characteristics of research coordinators (RCs) in a large pediatric emergency medicine research collaborative network. Our secondary objective was to determine if the RCs perceived any impact of those characteristics on their duties. METHODS/STUDY POPULATION: We conducted a 15-question electronic survey of RCs at the member institutions of the Pediatric Emergency Care Applied Research Network (PECARN). A total of 74 potential respondents were identified and received the survey. RESULTS/ANTICIPATED RESULTS: Fifty-three surveys (71.6%) were completed. Most respondents identified as female; white; and not Hispanic or Latino. Fourteen respondents (26.4%) identified as underrepresented minorities in medicine (UIM), which is similar to the percentage of UIM among the general population (30%). Twenty-eight respondents (52%) felt that their race/ethnicity positively impacted recruitment efforts. Twenty-three respondents (43%) felt that their ability to speak a language other than English positively impacted recruitment efforts. Four female respondents felt that their gender hindered their recruitment activities and impacted their sense of belonging within the research team. DISCUSSION/SIGNIFICANCE: RCs felt that their backgrounds and attributes positively impacted subject recruitment. However, some female coordinators felt negatively impacted by their gender. Increasing diversity amongst clinical research professionals and incorporating team cultural humility practices, may help increase diversity among clinical research subjects.

OBJECTIVES/SPECIFIC AIMS: The purpose of this study is to characterize children with nonalcoholic fatty liver disease (NAFLD) living in the Southeastern United States. METHODS/STUDY POPULATION: This retrospective electronic medical record chart review was conducted on a random sample of 206 children identified with NAFLD. Patients were included if they met the following criteria: confirmed NAFLD through either an ultrasound or liver biopsy or clinical suspicion of fatty liver disease alongside elevated alanine aminotransferase (ALT) in the absence of other etiologies causing elevated transaminases. Patients were excluded if they had hepatitis or other documented liver disease. Data collected at initial presentation included age, gender, ethnicity, height, weight, body mass index (BMI), BMI percentile, blood pressure, HbA1c, aspartate aminotransferase (AST), ALT, γ-glutamyl transferase (GGT), total cholesterol, total triglycerides, low-density lipoprotein, and high-density lipoprotein. Statistical analysis: for descriptive statistics, frequency counts and percentages alongside means, standard deviation, range, min/max values for the continuous variables were calculated. RESULTS/ANTICIPATED RESULTS: This study included 206 children diagnosed with NAFLD. Subjects were primarily male (n=136, 66%) and Caucasian (n=133, 66%), followed by Hispanic (n=42, 21%), Black (n=25, 12%), and Asian (n=2, 1%). Mean age at diagnosis was 12.3±3.5 years. Mean weight (lbs), height (in), and BMI (kg/m2) of subjects at diagnosis were 192±77 lbs, 61.7±6.6 in, 34.6±9.7 kg/m2, respectively. Patients had an average systolic blood pressure of 124±15.4 mmHg and diastolic blood pressure of 69.6±10.6 mmHg. Mean ALT was 91.8±67.2 U/L, AST was 61±38.8 U/L, and GGT was 55.1±64.6 U/L. Mean HbA1c was 5.8±1.4%, cholesterol was 176±36.3 mg/dL, triglycerides were 200±134 mg/dL, low-density lipoprotein was 107.6±32.1 mg/dL, and high-density lipoprotein was 39.9±8.4 mg/dL. DISCUSSION/SIGNIFICANCE OF IMPACT: In addition to having significantly elevated liver enzymes, children with NAFLD had several derangements in their metabolic profile, most notably high triglyceride levels and HbA1c values in the prediabetic range. Although lifestyle modification is the gold standard treatment for NAFLD, pharmacotherapy may need to be included to address metabolic syndrome.

OBJECTIVES/SPECIFIC AIMS: Prostate cancer (PCa) is the most common noncutaneous malignancy in men in the United States. A significant fraction of advanced PCa treated with androgen deprivation therapy experience relentless progression to lethal metastatic castration-resistant prostate cancer (mCRPC). The PCa tumor microenvironment is comprised of a complex mixture of epithelial and stroma cell types engaged in multifaceted heterotypic interactions functioning to maintain tumor growth and immune evasion. We recently uncovered the important role played by myeloid-derived suppressor cells (MDSCs) to mediate tumor immune evasion in aggressive PCa (Wang, Lu et al., Cancer Discovery, 2016). Immune checkpoint blockade (ICB) has elicited durable therapeutic responses across a number of cancer types. However, the impact of ICB on mCRPC has been disappointing, which may signal the need to combine mechanistically-distinct ICB agents and/or override immunosuppression in the tumor microenvironment. Our objective is to determine if robust immunotherapy responses in mCRPC may be elicited by the combined actions of ICB agents together with targeted agents that neutralize MDSCs yet preserve T cell function. METHODS/STUDY POPULATION: We created a novel embryonic stem cell-based chimeric mouse model of mCRPC engineered with signature mutations to study the response to single and combination immunotherapy. The efficacy studies were followed with detailed mechanistic investigation and clinical validation. RESULTS/ANTICIPATED RESULTS: Consonant with early stage clinical trials experience, anti-CTLA4 or anti-PD1 monotherapy failed to impact disease progression. Similarly, modest anti-tumor activity was observed with combination ICB as well as monotherapy with targeted agents including Cabozantinib (tyrosine kinase inhibitor), Dactolisib (PI3K/mTOR inhibitor), and Dasatinib (tyrosine kinase inhibitor). In contrast, mCRPC primary and metastatic disease showed robust responses to dual ICB treatment together with either Cabozantinib or Dactolisib, but not with Dasatinib which impaired T cell infiltration in the tumor. Detailed intratumoral immune profiling with mass cytometry (CyTOF) showed that combined ICB and Cabozantinib or Dactolisib was associated with significant depletion of MDSCs. Cabozantinib and Dactolisib blocked the PI3K signaling activity in MDSCs and reduced their immunosupppresive activity. Mechanistically, the combination efficacy was due to the upregulation of IL-1RA and suppression of MDSC-promoting cytokines secreted by mCRPC cells. DISCUSSION/SIGNIFICANCE OF IMPACT: We demonstrated that an antibody cocktail targeting CTLA4 and PD1 was insufficient to generate effective anti-tumor response, but combination of ICB with targeted therapy that inactivates PI3K signaling displayed superior synergistic efficacy through impairing MDSCs in the tumor microenvironment. These observations illuminate a clinical path hypothesis for combining ICB with MDSC-targeted therapies in the treatment of mCRPC. Importantly, conclusions from the study on PCa may have implications in combination immunotherapy for aggressive breast cancer which is also largely resistant to immune checkpoint blockade and replete with immunosuppressive myeloid cells.

ABSTRACT IMPACT: The car telemetry system may be an ideal method to accurately and reliably evaluate and compare at-risk driving errors between older drivers with and without glaucoma. OBJECTIVES/GOALS: Our project aims to determine whether an in-car telemetry system used during an on-road driving evaluation can accurately and reliably evaluate driving errors in lane maintenance and visual scanning and objectively quantify the frequency and severity of these errors in glaucoma patients. METHODS/STUDY POPULATION: This is a single center, cross-sectional study of 180 participants (125 with glaucoma and 55 controls), ages 55 or older, who underwent a comprehensive clinical assessment, including vision, cognition, motor function, followed by an on-road evaluation by a trained occupational therapist. Driving errors were recorded through a dual method including: 1. An in-car trained occupational therapist 2. In-car telemetry system. The frequency and severity of errors in lane maintenance and visual scanning from the in-car telemetry will be assessed and compared between participants with varying severity of glaucoma and normal controls. In addition, we will compare the frequency and severity of errors in lane maintenance and visual scanning to those recorded by the in-car driving evaluator. RESULTS/ANTICIPATED RESULTS: We anticipate (or predict) that the in- car telemetry system will be able to detect frequency and severity of driving errors in lane maintenance and visual scanning in glaucoma participants. We also predict that participants with worsening glaucoma severity will commit more driving errors. In addition, the in-car telemetry will detect a similar frequency and severity of driving errors as the in-car driving evaluator. DISCUSSION/SIGNIFICANCE OF FINDINGS: The type and frequency of vision-related driving errors that place individuals at risk for a car accident is not well known. Without this critical information, it is extremely challenging to help older adults with glaucoma to be safe drivers.

OBJECTIVES/GOALS: Parental SES may influence the trajectory of students applying to orthopaedic surgery residency, perpetuating opportunistic disparities. Thus, we sought to examine the relationship between parental occupation/education and applicant match rate, education financing, and medical school background. METHODS/STUDY POPULATION: Data from the Association of American Medical Colleges (AAMC) documented parental occupation and education levels of 10,697 orthopedics applicants from 2011 to 2021. Parental occupations were categorized into physician vs non-physician, healthcare vs non-healthcare, working class vs non-working class, and STEMM (Science, Technology, Engineering, Mathematics, Medicine) vs non-STEMM. Parental education levels spanned from no college degree to doctorate degrees and were used as a proxy for SES. Outcomes analyzed included match success, premedical and medical school debt, total educational debt, scholarships, and representation from top 40 research medical schools as determined by NIH funding. RESULTS/ANTICIPATED RESULTS: Physician parent applicants (20.1%) had better match rates (75.5% vs. 73.5%), lower debts, lesser scholarships, and higher top 40 school representation. Healthcare parent applicants (37.0%) had similar match rates, less debt and scholarships. Working class parent applicants (6.0%) had more debt and scholarships. STEMM parent applicants (48.6%) had higher match rates, lesser debts and scholarships, and higher top 40 representation. Applicants with parents without college degrees had lower match rates (68.6% vs 74.5%), more debt and scholarships. Doctorate parent applicants had better match success (75.9% vs 72.9%), lesser debts, and higher top 40 school representation (34.9% vs 29.6%). DISCUSSION/SIGNIFICANCE: Parental SES was associated with substantial variation in applicant financial burden and educational pedigree. Notably, applicants with parents lacking degrees had lower match rates, underscoring the need for supportive strategies to ensure equitable opportunities for aspiring orthopaedic surgeons.