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: This study examines faculty mentor experiences in a summer research program for students traditionally underrepresented in translational research. The objectives are to understand mentor perspectives of the program and how their views were impacted by the pandemic-related pivot to a remote format. METHODS/STUDY POPULATION: During the summer research program, students work with a faculty mentor on an ongoing research project. Program staff pair students with diverse faculty members who share their research interests. Our program has traditionally been offered as a residential in-person program but in 2020 we moved swiftly to a fully remote learning format. Students continued to work on research projects remotely and interacted virtually with their faculty mentors. For the past five cohort years, we have collected comparable data about faculty perspectives of their program experience, which allows us to evaluate the impact of the remote format on the faculty experience compared to that of the in-person format. RESULTS/ANTICIPATED RESULTS: For this study, we will examine data from five cohort years (2017-2021). A survey questionnaire was administered to mentors each year at the end of the summer research program. Data were collected on faculty satisfaction with the quality and amount of student work, the amount of time students spent on their projects, and how mentors communicated with students. In 2020 and 2021, three questions were added regarding satisfaction with the remote format. Quantitative data collected from both the in-person and remote cohorts will be compared using independent samples t-tests. Select quotes from open-ended qualitative questions will be used to illustrate mentors attitudes toward the program. DISCUSSION/SIGNIFICANCE: This study addresses a gap in the literature about summer research programs, as previous work has primarily focused on trainees. As many training programs continue to remain remote or adopt hybrid models, these results have implications for the design of similar mentored research programs.

OBJECTIVES/GOALS: The “hidden curriculum” is a set of unofficial rules outside of the formal curriculum that allows medical students to succeed. It is often not accessible to those who are first-generation in medicine. This study created a novel survey tool to directly evaluate the hidden curriculum, its contributing factors, and its effects on students. METHODS/STUDY POPULATION: Using available literature as a guide, a novel survey tool to evaluate different aspects of the hidden curriculum was created. This survey consists of 17 Likert scale questions on topics varying from sense of belongingness to dress code, self-guided studying, mentorship, and confidence in knowing how to succeed. This survey tool was embedded into a larger survey evaluating health disparities and diversity, inclusion, accessibility, and justice (DEIAJ) in the curricular and extracurricular spaces. This survey packet was administered to all medical students at a large U.S. medical school. RESULTS/ANTICIPATED RESULTS: 166 medical students from all years responded to this survey. 70% were female, 27% male, and 3% non-binary or prefer not to say. 67% of respondents agreed or strongly agreed that there is a hidden curriculum accessible to only those who have family members in medicine. 57% agreed or strongly agreed that the medical school gave them the adequate training and resources to succeed. 48% agreed or strongly agreed that they would perform better academically if they had more money with 11% stating they often feel embarrassed in a professional setting due to lack of money. Fellow classmen, faculty members, and upperclassmen were identified as the most useful resources to learn how to succeed in medical school. Students on average reported feeling like they knew what to do to succeed in medical school half of the time. DISCUSSION/SIGNIFICANCE: This data strongly supports the existence of a hidden curriculum and gives insight into the importance of financial support for low-income students and peer support groups for those who do not have family members in medicine. This data will be used to inform future interventions to address the hidden curriculum.

ABSTRACT IMPACT: Plasma and CSF are not reliable estimates of drug exposure in tissue compartments relevant for treatment and prevention of infectious diseases. OBJECTIVES/GOALS: Globally, high dose fluconazole is widely used in the management of cryptococcal meningitis. While it is known to readily penetrate into cerebrospinal (CSF), less is known about drug concentrations in brain parenchymal tissues. Similarly, distribution of fluconazole into gynecological tissues has not been robustly characterized. METHODS/STUDY POPULATION: With informed consent from next-of-kin, we conducted autopsies within 24h of death for hospitalized Ugandans receiving fluconazole for treatment or secondary prophylaxis of cryptococcal meningitis. Dosing history was abstracted from medical chart and caregiver interviews. Fluconazole concentrations were determined using high-performance liquid chromatography- tandem mass spectrometry (LC-MS/MS) in plasma, CSF, 10 brain compartments (frontal, parietal, and occipital lobes, corpus callosum, globus pallidus, hippocampus, midbrain, medulla oblongata, spinal cord, and choroid plexus) and 4 female genital compartments (cervix, vagina, ovary, and uterus), depending on tissue availability. Descriptive statistics of tissue to plasma ratios were used to describe concentrations relative to plasma. RESULTS/ANTICIPATED RESULTS: Fluconazole concentrations were measured in available tissues of 21 individuals with detectable fluconazole in plasma. Daily doses of fluconazole were 200 mg (n=4), 400 mg (n=1), 800 mg (n=4), 1200 mg (n=9) or unknown (n=3). CSF concentrations (n=10) ranged from 93-1380% (median 100%) of plasma while brain concentrations (n=3) across all 10 compartments ranged from 45% to 89% (median 69%) of plasma. In the female genital tract, cervical concentrations (n=10) were 9-78% (median 65%) of plasma and in the 2 individuals with available tissue, concentrations in vaginal, ovarian, and uterine tissues were similar to cervix, ranging from 63-105% of plasma. DISCUSSION/SIGNIFICANCE OF FINDINGS: Measuring drug concentrations directly in tissues, the presumed site of action, improves estimates of drug efficacy. While fluconazole concentrations in CSF were similar to plasma, actual brain tissues were consistently lower. Concentrations were similar between upper and lower female genital tracts, but were consistently lower than plasma.

OBJECTIVES/GOALS: To determine the prevalence of myocardial diastolic dysfunction (DD) and association of serum concentration of the cardiac biomarker serum soluble ST2 in HIV-infected as compared to uninfected Tanzanian adults at the time of HIV diagnosis. METHODS/STUDY POPULATION: In this cross-sectional study we consecutively enrolled HIV-infected participants and uninfected controls at a large, referral HIV clinic in Mwanza, Tanzania. Standardized history, physical examination, echocardiography and serum samples were obtained. The primary outcome was prevalence of myocardial diastolic dysfunction in HIV-infected as compared to uninfected adults. The secondary outcome was the association of baseline serum sST2 concentration with diastolic dysfunction prevalence. Regression models were used to quantify the associations. RESULTS/ANTICIPATED RESULTS: We enrolled 388 HIV-infected, ART naïve and 461 HIV-uninfected controls. Participants with HIV had a higher prevalence of DD (OR = 2.44, p = 0.001, controlled for age, sex, hypertension and BMI) and more severe dysfunction (66.7% vs 42.5%, p = 0.056) at an earlier age. Baseline serum sST2 concentration was significantly associated with DD in HIV-infected but not uninfected participants (p = 0.04 and 0.90, respectively). More HIV-infected adults with concurrent DD exceeded the threshold of 35ng/mL as compared to controls (15.7% vs 5.3%, p<0.0001). Additionally, a significant population level shift to higher sST2 concentration was observed in HIV-infected adults with dysfunction as compared to both HIV-infected without and HIV-uninfected adults with dysfunction (Kolmogrov-Smirnov test: p = 0.02 and 0.04). DISCUSSION/SIGNIFICANCE OF IMPACT: In a large population of HIV-infected adults in sub-Saharan Africa, HIV infection is associated with myocardial diastolic dysfunction. This dysfunction is associated with higher sST2 concentrations. Therefore, we conclude that the sST2 pathway may provide insight into the pathophysiologic mechanisms of dysfunction in HIV-infected adults.

OBJECTIVES/GOALS: Patients suffering from respiratory failure have few options to support oxygenation and carbon dioxide removal aside from mechanical ventilation. Our objective was to test a novel extrapulmonary mechanism of gas exchange via peritoneal oxygenated perfluorocarbon (PFC) in a large animal model. METHODS/STUDY POPULATION: Using two 50 kg swine, hypoxia was modeled with subatmospheric oxygen and hypercarbia induced with acute hypoventilation. Through a midline laparotomy, cannulas were placed into the peritoneal space to allow for PFC infusion and circulation. After abdominal closure, these cannulas were connected to a device capable of draining, oxygenating, and infusing PFC. One animal was subjected to acute hypoxia (12% FiO2) and another animal to acute hypoventilation (4 breaths per minute). Primary outcomes were times for SpO2 to reach 75 mmHg, respectively. Trials were performed without PFC and with PFC dwelling or circulating through the peritoneal space, during which abdominal and bladder pressures were monitored and maintained under 20 mmHg by regulation of the PFC volume contained in the animal. RESULTS/ANTICIPATED RESULTS: In the animal subjected to acute hypoxia (12% FiO2), survival time improved from 5:55 to 20:00 (min:sec) after 2.5 liters of oxygenated PFC was instilled in the peritoneal space. Oxygen percent saturation of PFC before and after dwelling in the peritoneal space was measured at 100% before and 70% after dwelling in the animal during this hypoxic period corresponding with a gas transfer of 300 mL of oxygen over the 20-minute trial (i.e., 15 mL/min). Continual PFC circulation did not further extend the survival time during hypoxic conditions over PFC dwelling in the abdomen. In the animal that was acutely hypoventilated, there were no detectable differences in the rate of CO2 accumulation as measured by EtCO2 or direct blood pCO2 measurements with PFC dwelling or circulating through the peritoneal space. DISCUSSION/SIGNIFICANCE: Oxygenated PFC dwelling in the peritoneal space increased the duration of systemic arterial blood saturation remaining greater than 50% during normobaric hypoxic (12% FiO2) conditions but did not appreciably clear blood carbon dioxide during hypoventilation. Future experiments will focus on maximizing the rate of systemic oxygen uptake.

OBJECTIVES/GOALS: The desmoplastic reaction in PDAC involves a significant accumulation of immune cells and fibroblasts.The functional diversity of carcinoma associated fibroblasts (CAFs) remains largely unknown, and identification of immune regulating subsets would have a substantial impact in augmentation of immunotherapy efficacy. METHODS/STUDY POPULATION: Employing histology, FACs, multiplex immunohistochemistry, single cell RNA sequencing (sc-RNA-seq) and genetically engineered mouse models, we demonstrate that aSMA+ cells are a dominant CAF population in PDAC with tumor restraining properties (TS-CAFs), as opposed those of the FAP+ CAFs, which demonstrate tumor promoting activity (TP-CAFs). RESULTS/ANTICIPATED RESULTS: Analysis of bulk tumor depleted of either TS-CAFs or TP-CAFs showed that TS-CAFs predominantly modulate extracellular matrix (ECM) production, facilitate cell-ECM adhesion and regulate adaptive immunity, while TP-CAFs exhibit a lineage that is skewed towards a pro-inflammatory, chemokine secreting phenotype. Further, scRNA-Seq analyses demonstrate that CAFs share distinct gene expression profiles characteristic of lymphocytic and myeloid lineages. Together our data distinguish two populations of CAFs, one which is tumor suppressing with roles in ECM remodeling and another which is tumor promoting with roles in cytokine production, both with immune modulating capabilities. DISCUSSION/SIGNIFICANCE OF IMPACT: Our study identifies a complex network of functionally heterogeneous fibroblasts during PDAC progression with significant immunotherapeutic implication. The identification of distinct fibroblast subsets will allow us to discriminately target fibroblast populations to augment immunotherapy efficacy in pancreatic cancer.

ABSTRACT IMPACT: This work represents a novel way in which genetic information can be used to improve clinical decision making as it pertains to both treatment and management of congenital heart disease. OBJECTIVES/GOALS: Our lab found that MYH6 variants are both enriched in hypoplastic left heart syndrome (HLHS) and associated with decreased cardiac transplant-free survival. To elucidate the mechanisms of MYH6 variant pathogenicity, we are assessing their impact on atrial function during HLHS development and progression. METHODS/STUDY POPULATION: We are using 2D speckle-based tracking to retrospectively evaluate echocardiograms (echos) from 51 HLHS patients, 17 with MYH6 variants and 34 matched controls. Atrial function will be assessed by myocardial strain and strain rate at seven time points, beginning at the time of the patients’ earliest prenatal echo, and ending with their last available echo before death or cardiac transplant. Early left atrial function will examine the role of MYH6 variants in the development of HLHS in vivo, while longitudinal right atrial function will be assessed in order to look for differences that could be contributing to the decreased transplant-free survival seen in MYH6 variant carriers. RESULTS/ANTICIPATED RESULTS: We hypothesize that MYH6 variants cause HLHS by impairing early left atrial (LA) contractility, resulting in altered left ventricular hemodynamics and consequent hypoplasia. We therefore expect to find diminished prenatal LA function in HLHS patients with MYH6 variants. We also hypothesize that MYH6 variants continue to impair right atrial (RA) function in surgically-reconstructed HLHS hearts, necessitating earlier transplantation. Accordingly, we expect variant carriers to exhibit lower RA function at birth versus controls. We expect differences between groups to persist over time, and possibly increase in magnitude. In HLHS patients with MYH6 variants, we anticipate declining RA function will precede right ventricular function and therefore be an early indicator of transplant need. DISCUSSION/SIGNIFICANCE OF FINDINGS: This study represents a novel way in which genetic information can inform clinical decision-making. Identifying MYH6 variants as an early cause of HLHS offers chances for intervention. Understanding long-term effects of MYH6 on right atrial function in HLHS may aid in cardiac transplant risk stratification, thus improving patient outcomes.

OBJECTIVES/GOALS: A stroke can impair neural communication between sensory and motor pathways thus compromising walking function. Paired associative stimulation (PAS) is a useful assay of sensorimotor integration (SMI) with limited use post-stroke. The objective of this study will be to determine lower extremity PAS effectiveness and reliability post-stroke. METHODS/STUDY POPULATION: This study will use a pre-post, cross-sectional design. Ten healthy controls and 10 individuals with chronic stroke (>6 months) will be recruited. PAS protocols will be individualized to account for between-subject variability in sensorimotor signaling by first measuring cortical sensory signaling using electroencephalography. Post-stroke participants will then receive PAS targeting the paretic tibialis anterior muscle; healthy controls will receive PAS targeting the non-dominant TA. Changes in cortically derived muscle responses will be characterized by absolute motor-evoked potential amplitude (MEPAmp) change, elicited by transcranial magnetic stimulation, over two sessions separated by >24 hours. Clinical measures of sensorimotor function and walking ability will also be performed. RESULTS/ANTICIPATED RESULTS: By individualizing PAS protocols, we expect to see significant increases in MEPAmp pre to post PAS, determined using paired t-tests. We also anticipate reliable PAS-induced increases in MEPAmp, which will be assessed using two reliability statistics: intraclass correlation coefficient and coefficients of variation of method error. Lastly, the increases in MEPAmp will be correlated with measures of sensorimotor function and walking ability, anticipating that greater increases in MEPAmp will be related to better walking ability and sensorimotor functioning. Correlations will be assessed via a Pearson’s correlation. A preset alpha = 0.05 will be used to determine significant findings. DISCUSSION/SIGNIFICANCE: The importance of this study is that establishing individualized PAS protocols could potentially provide a reliable and clinically relevant measure of SMI. Understanding post-stroke lower extremity SMI is necessary for furthering targeted and personalized interventions to combat walking deficits.

OBJECTIVES/SPECIFIC AIMS: o To review the community’s recommendations on how to rebuild trust in the Flint community. o To review effective community engagement strategies utilized with the Flint Special Projects for project conceptualization, participant recruitment, data analysis, project oversight, and dissemination. METHODS/STUDY POPULATION: The study population includes nearly two hundred residents representing seniors, youth and diverse ethnicities recruited to participate in eleven focus group meetings. The population also represents the general public who attended informational meetings in Flint, Michigan to learn about the crisis and allow residents to voice their opinions and concerns during the onset of the crisis. The project is a mixed methods community based participatory research effort that utilized community decision making in all phases of the effort such as pre-conception, implementation, dissemination and advocacy to encourage the community’s recommendations are adopted at policy and institutional responsiveness levels. It includes three community engaged research efforts: (project 1) A qualitative analysis of community sentiment provided during 17 recorded legislative, media and community events, and (projects 2-3) two mixed methods efforts utilizing purposive sampling of stakeholders whose voice may not have been heard. RESULTS/ANTICIPATED RESULTS: The project presents a qualitative analysis of the community’s voice during the onset of the man-made disaster when the community first became aware of the emergency manager’s plans to switch the water source. It also reflects current perspectives of community voice since the projects are scheduled to end late February 2019. Findings from a trust measure administered to nearly two hundred residents will be presented, along with a qualitative analysis of focus group findings among segments of the population (seniors, youth, and diverse ethnicities) who may have been left out of narratives on the water crisis. Finally, the project will compare empowerment and resiliency approaches being utilized in Flint, Michigan to recover from the disaster with other approaches grounded in literature and theory. DISCUSSION/SIGNIFICANCE OF IMPACT: Communities of color often experience social determinants of health which negatively impact their health, well-being and human rights. Some Flint citizens are experiencing negative health consequences (i.e., rashes, brain and behavioral sequelle, fertility, etc.) as a result of the disaster, and are uncertain of health outcomes in the future. This is the first project to rigorously document and analyze levels of trust and mistrust in the city of Flint since the water disaster occurred. The qualitative research will guide future clinical research that will benefit this traumatized community experiencing high levels of mistrust (i.e., government, elected officials, etc.). The community engaged methodology involved residents and study participants in all phases of the project including project oversight, validating and analyzing data, and dissemination. This methodology will contribute to existing literature and theory on community based participatory research, community engaged research, team science and citizen science. The approaches empowered a call to action among residents, for example, seniors who attended two senior focus group sessions shared “they are hopeful and have a purpose,” resulting in the creation of a council (with officers) at their housing complex to advocate for the well-being of seniors during the recovery process. Recruitment methodologies were extremely successful due to resident level trust in community leaders and community partner organizations. Finally, the project’s examination of approaches encouraging empowerment and resiliency will provide lessons learned for other communities challenged with crisis.

OBJECTIVES/GOALS: Thiswork is an evidential study that demonstrates the positive impactof integrating Community Health Workers (CHWs) and SocialDeterminants of Health on an important health outcome, notably in decreasing the 30-day unplanned hospital ED readmissions at Sinai Health Systems. METHODS/STUDY POPULATION: Using datafrom the Sinai Urban Health Institute (SUHI), we compare predictingthe readmissions of patients with and without data pertainingto Social Determinants of Health (SDoH). We thoroughly describe the data cleaning and data pre-processing, done in collaboration with experts in community health. We use a fundamental and ubiquitous classifier in Random Forest for its feature characterization capability in order to translate models results into insights and recommendations for the CHW program. RESULTS/ANTICIPATED RESULTS: We show that when patients are simply engaged byCHWs, regardless of the content of those conversations, we canincrease the predictive accuracy of our classifier by 5%. We usethis result to make recommendations for improving patient careand discuss limitations and future work. Importantly our workpoints directly to the human connection between patients andCHWs as an important feature in the readmission rate. DISCUSSION/SIGNIFICANCE: Our work shows that the predictive capabilities of the classifier increases with CHW logs and SDoH survey data, highlighting the benefit of collecting this information. This is the first step in early identification of such patients so that CHWs are focusing on and providing resources to patients who will most benefit from the program.

OBJECTIVES/SPECIFIC AIMS: To examine maternal morbidity and its related social determinants among women experiencing homelessness during pregnancy. METHODS/STUDY POPULATION: This study will use an exploratory sequential mixed method design to explore and examine the structural, interpersonal and individual factors contributing to maternal morbidity among a convenience sample of 150 English speaking women experiencing homelessness during a pregnancy within the last 3 years in Baltimore. In the qualitative phase of the study, we will conduct semi-structured interviews with 15 women purposively sampled to refine the relationships between resilience, social determinants of health and multilevel factors that impact maternal morbidities. Factors of interest include prenatal care received, barriers and facilitators to receiving prenatal care, maternal morbidities, social support, and strategies used to manage their condition during this time. Using the findings from the qualitative phase, a quantitative survey will be developed to gather data on topics that emerged in the interviews. In addition, the Housing Instability Index will be used to measure the degree of homelessness as defined by the degree of housing instability in a 6-month period. Using the 25-item Connor-Davidson Resilience Scale, resilience levels among women in the sample will be assessed as a moderating factor in the examination of the relationship between a pregnant woman’s homeless status and maternal morbidity. Descriptive statistics and logistical regression tests will be used to analyze these relationships while controlling for other structural, interpersonal, and individual factors that may be associated with maternal morbidity. RESULTS/ANTICIPATED RESULTS: Qualitatively we expect to gain insight into the relationship between the extrinsic and intrinsic factors impacting maternal morbidities and the health behaviors and practices used by women to manage their pregnancy while homeless. These findings will inform the quantitative survey development and help generalize the quantitative findings. We expect to identify the common morbidities in this population we anticipate that there will be differences in maternal morbidity among the different types of homelessness. Maternal morbidity will be higher among women with a greater degree of homelessness. Resilience will have a moderating effect on the relationship between homelessness and maternal morbidity. DISCUSSION/SIGNIFICANCE OF IMPACT: This study, to our knowledge, is the first to look at maternal morbidity in this population. Additionally, this study seeks to move current research from examining infant outcomes at birth among mothers experiencing homelessness to understanding the maternal morbidities during this period. Long term, good maternal health has significant implications for the health of a mother’s future pregnancies and a risk reduction of adverse chronic conditions. Study results will provide the preliminary knowledge needed to guide further research leading to clinical approaches that promote better maternal health in this population. Lastly, the study findings will inform policy by characterizing the quality and strength of evidence of the adverse maternal health effects associated with the experience of homelessness.

ABSTRACT IMPACT: As the number of poorly water-soluble drugs in development increases, our research will expand on the science behind improving drug solubility and absorption and ensuring that promising poorly-water solubility drugs do not fail drug development. OBJECTIVES/GOALS: Spray-dried dispersion (SDD) tablet formulation is an approach to increase oral drug solubility and absorption. Methods to predict SDD performance in humans are poorly developed. We aim to develop an in vivo in vitro correlation (IVIVC) between in vitro dissolution and in vivo absorption of itraconazole SDD tablets. METHODS/STUDY POPULATION: This research project involves tablet manufacturing, in vitro dissolution experiments, and a clinical study. We manufactured fast-, medium-, and slow-release SDD tablets containing amorphous solid dispersion of itraconazole (100 mg) and different grades of the polymer hypromellose acetate succinate (HPMC-AS). Tablets differed in slug pressure, tablet compression force, and formulation composition. Dissolution studies were performed using the United States Pharmacopeia (USP) type II apparatus. The clinical study is an ongoing randomized, cross-over, open-label, fasted, single-dose trial in healthy participants (n=12). An IVIVC will be created by comparing the rank order of drug in vitro dissolution with in vivo absorption. RESULTS/ANTICIPATED RESULTS: Tablet manufacturing was successful, and the tablets displayed the same dissolution rate ranking order as anticipated. Fast-release tablets showed the highest percentage of drug dissolved by 10 min (74%) compared to medium- (62%) and slow-release (1.2%) tablets. Percentage drug dissolved differs by at least 10% at all time points among the different release-rate tablets. The clinical study is currently ongoing, and we expect that the pharmacokinetic (PK) profiles differ among the different tablets. We predict that the rank order of tablet absorption in humans will agree with the order of drug dissolved observed in the dissolution experiments. DISCUSSION/SIGNIFICANCE OF FINDINGS: Spray-dried dispersions are a formulation method to try to improve drug solubility and oral drug absorption. This research will elucidate manufacturing parameters that can impact tablet performance and expand on the ability of in vitro dissolution to predict human PK and streamline drug development of poorly soluble drug candidates.

OBJECTIVES/GOALS: The goal was to develop foundational research knowledge and skills for clinicians interested in conducting clinical research. Emphasis was on the development of a research question and the iterative process necessary to transform a research question into a well-designed study and well-articulated research proposal for pilot grant funding. METHODS/STUDY POPULATION: The course took place over 10 sessions, May - September 2021. The application process required participants to provide an initial self-assessment of research skills and a proposed research question. 25 clinicians applied to the program, 11 were accepted, and 9 enrolled. All clinicians in the initial cohort were clinical faculty physicians. Because of the geographical distance of participants as well as the ongoing pandemic, the course used a blended learning approach with both synchronous and asynchronous learning. Participants viewed online lectures on core content coupled with live virtual sessions with opportunities for discussion and application of the content. Relevant CTSI and institutional resources were highlighted in each session. RESULTS/ANTICIPATED RESULTS: Of the 9 clinician participants who enrolled in Research 101, the average attendance per session was 67% or 6 people. 89% or 8 participants attended five or more sessions. 5 participants submitted a letter of intent for the annual CTSI Pilot grant program, and of the five, two were invited to submit a grant application. Formal evaluation of the program is currently ongoing and will close on November 30th, at which time full results will be available. Research competencies will be assessed through a pre-post comparison, each self-rated by course participants. Additionally, participants were asked to provide input on the most and least valuable components of the course, as well as any open-ended feedback. Research 101 leadership will use these results to improve the course for future participants. DISCUSSION/SIGNIFICANCE: A learning health system (LHS) is recognized as an essential means by which research evidence is translated into practice. Important to realizing the LHS vision is the engagement of clinicians into the generation and translation of research into practice. Research 101 is an important way to bolster clinician engagement in translational research.

OBJECTIVES/GOALS: The Clinical Implementation stage in the translational pipeline is hampered by the tension between formal evidence and clinician perceptions. For instance, when guidelines are translated into electronic clinical decision support alerts, they are often ignored. Using advances in LLMs we present a framework to quantify these discrepancies. METHODS/STUDY POPULATION: We hypothesize that ignoring guideline-based alerts may be driven by discordances between clinical guidelines’ deterministic realities and clinician’ perception of clinical reality. Until now this has been very difficult to measure using quantitative methods. We argue that advances in Large Language Models (LLM) provide an avenue for exploring this quantitatively. Here we present the method and preliminary results comparing the responses of BioBERTT from a carefully designed set of questions when the LLM is fine-tuned using either formal guidelines or transcripts of clinicians discussing guidelines and clinical care in the parallel domain. The formal “distance” between the LLM responses is evaluated using quantitative metrics like the Hamming Distance. RESULTS/ANTICIPATED RESULTS: We present a description of the architecture used to prove or disprove our hypothesis. We will present results obtained when training the architecture with data that could be used to test the limits of our hypothesis, by fine-tuning BioBERT with diverse synthetic clinical views, either in agreement or disagreement with the formal guidelines. Results comparing sepsis guideline text with transcripts of interviews with Emergency Department clinicians discussing care practices for sepsis in the ED transcripts will also be considered. Our current emphasis is on securing a wider range of transcripts of clinicians interviewed from different clinical specialties and different clinical settings. While here we focus on clinical guidelines, the framework supports any intervention in the Clinical Implementation stage. DISCUSSION/SIGNIFICANCE: Leveraging recent advances in LLMs, we develop a framework that can quantitatively measure the differences between guidelines and clinician perception of best practices. We demonstrated the functionality of this approach using synthetic data and initiated the collection of clinician transcripts to test the framework in real clinical situations.

Ensuring appropriate review, approval, and oversight of research involving animals becomes increasingly complex when researchers collaborate across multiple sites. In these situations, it is important that the division of responsibilities is clear and that all involved parties share a common understanding. The National Institutes of Health Office of Laboratory Animal Welfare and the United States Department of Agriculture Animal Plant Health Inspection Service require an Institutional Animal Care and Use Committee (IACUC) to review the care and use of animals in research, and both agree that it is acceptable for one IACUC to review the work taking place at multiple institutions. With this in mind, several Harvard-affiliated hospitals and academic centers developed the Master Reciprocal Institutional Agreement for Animal Care and Use (Master IACUC Agreement) to support collaboration, decrease administrative burden, increase efficiencies, reduce duplicative efforts, and ensure appropriate protections for animals used in research. Locally, the Master IACUC Agreement has fostered greater collaboration and exchange while ensuring appropriate review and oversight of research involving animals. As multisite animal protocols become more prevalent, this Agreement could provide a model for a distributed, national network of IACUC reliance.

The 2020 COVID-19 pandemic has had a profound impact on the clinical research enterprises at the 60 Clinical and Translational Science Award (CTSA) Hubs throughout the nation. There was simultaneously a need to expand research to obtain crucial data about disease prognosis and therapy and enormous limitations on conducting research as localities and institutions limited travel and person-to-person contact. These imperatives resulted in major changes in the way research was conducted, including expediting Institutional Review Board review, shifting to remote interactions with participants, centralizing decision-making in prioritizing research protocols, establishing biobanks, adopting novel informatics platforms, and distributing study drugs in unconventional ways. National CTSA Steering Committee meetings provided an opportunity to share best practices and develop the idea of capturing the CTSA program experiences in a series of papers. Here we bring together the recommendations from those papers in a list of specific actions that research sites can take to strengthen operations and prepare for similar future public health emergencies. Most importantly, creative innovations developed in response to the COVID-19 pandemic deserve serious consideration for adoption as new standards, thus converting the painful trauma of the pandemic into “post-traumatic growth” that makes the clinical research enterprise stronger, more resilient, and more effective.

Translation of research discoveries into health impact can take many years, creating delays in improving clinical outcomes. One approach to promoting timely translation is to examine successful cases in order to understand facilitators and strategies for overcoming barriers. We examined the development of evidence-based management for neonatal abstinence syndrome (NAS) at one academic medical center, with a primary focus on pharmacologic treatment. Despite a substantial increase in NAS case incidence starting in the early 2000s, significant sociocultural, policy, and regulatory barriers limited collaborative NAS research. Facilitators for translation encompassed: 1) pursuing research of societal interest, 2) building an effective interdisciplinary team, 3) intentionally linking clinical, research, and advocacy efforts, 4) broad stakeholder engagement across clinical, policy, and research arenas, and 5) leveraging local resources. Challenges included lack of commercially available U.S. Food and Drug Administration approved neonatal drug formulations, legal and regulatory barriers related to off-label and illicit use of opioids, recruitment for a treatment associated with drug withdrawal syndromes, misalignment of research design needs with real-world scenarios, and episodic funding. Benefits of successful translation included improvements in clinical care, reduced healthcare costs related to NAS, and enhanced legislative, policy, and research strategies to support broader neonatal investigations.

Adenylate kinases (AKs) are important enzymes involved in cellular energy metabolism. Among AKs, AK5 (adenylate kinase 5), a cytosolic protein, is emerging as a significant contributor to various diseases and cellular processes. This comprehensive review integrates findings from various research groups on AK5 since its discovery, shedding light on its multifaceted roles in nucleotide metabolism, energy regulation, and cellular differentiation. We investigate its implications in a spectrum of diseases, including autoimmune encephalitis, epilepsy, neurodegenerative disorders such as Alzheimer’s and Parkinson’s, diabetes, lower extremity arterial disease, celiac disease, and various cancers. Notably, AK5’s expression levels and methylation status have been associated with cancer progression and patient outcomes, indicating its potential as a prognostic indicator. Furthermore, AK5 is implicated in regulating cellular processes in breast cancer, gastric cancer, colorectal carcinoma, prostate cancer, and colon adenocarcinoma, suggesting its relevance across different cancer types. However, a limitation lies in the need for more robust clinical validation and a deeper understanding of AK5’s precise mechanisms in disease pathogenesis, despite its association with various pathophysiological conditions. Nonetheless, AK5 holds promise as a therapeutic target, with emerging evidence suggesting its potential in therapy development.