OBJECTIVES/GOALS: There is an interest in developing a bioprosthetic ovary for ovarian tissue transplantation. The properties of the ovarian extracellular matrix need to be better understood in order to replicate the human ovary. We tested the effects of an encapsulating hydrogel at different rigidities on bovine primordial follicle activation, growth, and survival. METHODS/STUDY POPULATION: Bovine primordial follicles were isolated from ovarian cortex. A mean of 9.9 follicles (range 3-24) were encapsulated per bead in either 1% or 5 % alginate across 4 experiments. The encapsulated follicles were subsequently crosslinked in a calcium sulfate solution. Follicles were then cultured for 8 days with light microscopy imaging taken every other day along with media exchanges. Follicles were then examined using immunofluorescence. Growth and survival curves were constructed and all statistical analyses were performed using Graph Pad Prism 9. RESULTS/ANTICIPATED RESULTS: A total of 372 follicles were encapsulated across 32 beads (16 in 1% alginate and 16 in 5% alginate). There were no differences in initial follicle size between the two conditions (33.53 µm vs. 32.45 µm, p=0.47). At the end of 8 days, there was no difference between follicle size (59.55 vs. 56.06, p=0.48). Additionally, there was no difference in survival between 1% and 5% alginate encapsulation (57.75% vs. 52.43%. p=0.40). Immunofluorescence is being performed on encapsulated follicles to confirm the presence of DDX4, a molecular marker of oocytes, after 8 days in culture. Additional encapsulated follicles have also been submitted for histologic sectioning and hematoxylin and eosin staining to better characterize the viability and health of these follicles after 8 days in culture. DISCUSSION/SIGNIFICANCE: There was no significant difference in growth or survival between primordial follicles cultured in 1% or 5% alginate gels. Immunofluorescent analysis confirmed the presence of viable follicles at the end of 8 days of culture. Future work needs to further explore how factors in the ovarian extracellular matrix impact follicle maintenance and growth.

OBJECTIVES/SPECIFIC AIMS: Stigma has been recognized as a majorimpediment to accessing mental health care among Vietnamese and Asian Americans(Leong and Lau, 2001; Sadavoy et al., 2004; Wynadenet al., 2005; Fong and Tsuang, 2007). The underutilizationof mental health care, and disparities in both access and outcomes have beenattributed to a large extent to stigma and cultural characteristics of thispopulation (Wynaden et al., 2005; Jang et al.,2009; Leung et al., 2010; Spencer et al.,2010; Jimenez et al., 2013; Augsberger et al.,2015). People with neurotic or behavioral disorders may be considered“bad” as many Vietnamese people believe it is aconsequence of one’s improper behavior in a previous life, for whichthe person is now being punished (Nguyen, 2003). Mental disorders can also beenseen as a sign of weakness, which contributes to ambivalence and avoidance ofhelp-seeking (Fong and Tsuang, 2007). Equally important is the need to protectfamily reputation; having emotional problems often implies that the person has“bad blood” or is being punished for the sins ofhis/her ancestors (Herrick and Brown, 1998; Leong and Lau, 2001),which disgraces the entire family (Wynaden et al., 2005). Inthese cases, public stigma (as opposed to internal stigma) is the primary reasonfor delays in seeking help (Leong and Lau, 2001). Other research has alsohighlighted the influences of culture on how a disorder may be labeled indifferent settings, although the presentation of symptoms might be identical(see Angel and Thoits, 1987). In Vietnamese culture, mental disorders are oftenlabeled điên (literally translated as“madness”). A điên person and his orher family are often severely disgraced; consequently the individuals and theirfamily become reluctant to disclose and seek help for mental health problems forfear of rejection (Sadavoy et al., 2004). Despite the criticalrole of stigma in accessing mental health care, there has been little work intrying to understand how stigmatizing attitudes towards mental illness amongVietnamese Americans manifest themselves and the influences of acculturation onthese attitudes. Some previous work indicated a significant level of mentalillness stigma among Vietnamese Americans, and experiences of living in theUnited States might interact with the way stigma manifests among this population(Do et al., 2014). Stigma is a complex construct that warrantsa deeper and more nuanced understanding (Castro et al., 2005).Much of the development of stigma-related concepts was based on the classic workby Goffman (1963); he defined stigma as a process by which an individualinternalizes stigmatizing characteristics and develops fears and anxiety aboutbeing treated differently from others. Public stigma (defined by Corrigan, 2004)includes the general public’s negative beliefs about specific groups,in this case individuals and families with mental illness concerns, thatcontribute to discrimination. Public stigma toward mental illness acts not onlyas a major barrier to care, but can also exacerbate anxiety, depression, andadherence to treatment (Link et al., 1999; Sirey etal., 2001; Britt et al., 2008; Keyes etal., 2010). Link and Phelan (2001) conceptualized public stigma throughfour major components. The first component, labeling, occurs when peopledistinguish and label human differences that are socially relevant, for example,skin color. In the second component, stereotyping, cultural beliefs link thelabeled persons to undesirable characteristics either in the mind or the body ofsuch persons, for example people who are mentally ill are violent. The thirdcomponent is separating “us” (the normal people) from“them” (the mentally ill) by the public. Finally, labeledpersons experience status loss and discrimination, where they are devalued,rejected and excluded. Link and Phelan (2001) emphasized that stigmatizationalso depends on access to social, economic, and political power that allowsthese components to unfold. This study aims to answer the following researchquestions: (1) how does public stigma related to mental illness manifest amongVietnamese Americans? and (2) in what ways does acculturation influence stigmaamong this population? We investigate how the 4 components of stigma accordingto Link and Phelan (2001) operationalized and how they depend on the level ofacculturation to the host society. Vietnamese Americans is the key ethnicminority group for this study for several reasons. Vietnamese immigration, whichdid not start in large numbers until the 1970s, has features that allow for anatural laboratory for comparisons of degree of acculturation. Previous researchhas shown significant intergenerational differences in the level ofacculturation and mental health outcomes (e.g., Shapiro et al.,1999; Chung et al., 2000; Ying and Han, 2007). In this study,we used age group as a proxy indicator of acculturation, assuming that those whowere born and raised in the United States (the 18–35 year olds) wouldbe more Americanized than those who were born in Vietnam but spent a significantpart of their younger years in the United States (the 36–55 yearolds), and those who were born and grew up in Vietnam (the 56–75 yearolds) would be most traditional Vietnamese. The language used in focus groupdiscussions (FGDs) reflected some of the acculturation, where all FGDs with theyoungest groups were done in English, and all FGDs with the oldest groups weredone in Vietnamese. METHODS/STUDY POPULATION: Data were collectedthrough a set of FGDs and key informant interviews (KIIs) with experts toexplore the conceptualization and manifestation of mental illness public stigmaamong Vietnamese Americans in New Orleans. Six FGDs with a total of 51participants were conducted. Participants were Vietnamese American men and womenages 18–75. Stratification was used to ensure representation in thefollowing age/immigration pattern categories: (1) individuals age56–75 who were born and grew up in Vietnam and immigrated to theUnited States after age 35; (2) individuals age 36–55 who were bornin Vietnam but spent a significant part of their youth in the United States; and(3) individuals age 18–35 who were born and grew up in the UnitedStates. These groups likely represent different levels of acculturation,assuming that people who migrate at a younger age are more likely to assimilateto the host society than those who do at a later age. Separate FGDs wereconducted with men and women. Eleven KIIS were conducted with 6 serviceproviders and 5 community and religious leaders. In this analysis, we focused onmental illness public stigma from the FGD participants’ perspectives.FGDs were conducted in either English or Vietnamese, whichever participants feltmore comfortable with, using semistructured interview guides. All interviewswere audio recorded, transcribed and translated into English if conducted inVietnamese. Data coding and analysis was done using NVivo version 11 (QSRInternational, 2015). The analysis process utilized a Consensual QualitativeResearch (CQR) approach, a validated and well-established approach to collectingand analyzing qualitative data. CQR involves gathering textual data throughsemistructured interviews or focus groups, utilizing a data analysis processthat fosters multiple perspectives, a consensus process to arrive at judgmentsabout the meaning of data, an auditor to check the work of the research team,and the development of domains, core-ideas, and cross-analysis (Hill etal., 2005). The study was reviewed and approved by TulaneUniversity’s Internal Review Board. RESULTS/ANTICIPATEDRESULTS: Components of public stigma related to mental illness. The 4 componentsof public stigma manifest to different extents within the Vietnamese Americansin New Orleans. Labeling was among the strongest stigma components, while theevidence of the other components was mixed. Across groups of participants,Vietnamese Americans agreed that it was a common belief that people with mentaldisorders were “crazy,” “actingcrazy,” or “madness.” “Notnormal,” “sad,” and“depressed” were among other words used to describe thementally ill. However, there were clear differences between younger and olderVietnamese on how they viewed these conditions. The youngest groups ofparticipants tended to recognize the “craziness” and“madness” as a health condition that one would need toseek help for, whereas the oldest groups often stated that these conditions wereshort term and likely caused by family or economic problems, such as a divorce,or a bankruptcy. The middle-aged groups were somewhere in between. The evidencesupporting the second component, stereotyping, was not strong among VietnameseAmericans. Most FGD participants agreed that although those with mentaldisorders may act differently, they were not distinguishable. In a few extremecases, mentally ill individuals were described as petty thefts or being violenttowards their family members. Similarly to the lack of strong evidence ofstereotyping, there was also no evidence of the public separating the mentallyill (“them”) from “us”. It wasnearly uniformly reported that they felt sympathetic to those with mentaldisorders and their family, and that they all recognized that they needed help,although the type of help was perceived differently across groups. The olderparticipants often saw that emotional and financial support was needed to helpindividuals and families to pass through a temporary phase, whereas youngerparticipants often reported that professional help was necessary. The lastcomponent, status loss and discrimination, had mixed evidence. While nearly noparticipants reported any explicit discriminatory behaviors observed andpracticed towards individuals with mental disorders and their families, wordslike “discrimination” and “stigma”were used in all FGDs to describe direct social consequences of having a mentaldisorder. Social exclusion was common. Our older participants said:“They see less of you, when they see a flaw in you theydon’t talk to you or care about you. That’s one thing theVietnamese people are bad at, spreading false rumors anddiscrimination” (Older women FGD). One’s loss of statusseemed certain if their or their loved one’s mental health status wasdisclosed. Shame, embarrassment, and being “frowned upon”were direct consequences of one’s mental health status disclosure andsubsequently gossiped about. Anyone with mental disorders was certain toexperience this, and virtually everyone in the community would reportedly dothis to such a family. “You get frowned upon. In the Vietnameseculture, that’s [a family identified as one with mentalhealth problems] the big no-no right there. When everybody frownsupon your family and your family name, that’s when it becomes aproblem” (Young men FGD). This is tied directly to what ourparticipants described as Vietnamese culture, where pride and family reputationwere such a high priority that those with mental disorders needed to go to agreat extent to protect—“We all know what saving facemeans” as reported by our young participants. Even among youngparticipants, despite their awareness of mental illness and the need forprofessional help, the desire to avoid embarrassment and save face was so strongthat one would think twice about seeking help. “No, you justdon’t want to get embarrassed. I don’t want to go to thedamn doctor and be like ‘Oh yeah, my brother got an issue. You canhelp him?’ Why would I do that? That’s embarrassing tomyself…” (Young men FGD). Our middle-aged participantsalso reported: “If I go to that clinic [mental health orcounseling clinic], I am hoping and praying that I won’tbump into somebody that I know from the community” (Middle-aged womenFGD). Vietnamese people were also described as being very competitive amongthemselves, which led to the fact that if a family was known for having anyproblem, gossips would start and spread quickly wherever they go, and prettysoon, the family would be looked down by the entire community. “Ithink for Vietnamese people, they don’t help those that are in need.They know of your situation and laugh about it, see less of you, and distantthemselves from you” (Older women FGD). Culture and mental illnessstigma, much of the described stigma and discrimination expressed, andconsequently the reluctance to seek help, was attributed to the lack ofawareness of mental health and of mental health disorders. Many studyparticipants across groups also emphasized a belief that Vietnamese Americanswere often known for their perseverance and resilience, overcoming wars andnatural disasters on their own. Mental disorders were reportedly seen asconditions that individuals and families needed to overcome on their own, ratherthan asking for help from outsiders. This aspect of Vietnamese culture isintertwined with the need to protect one’s family’sreputation, being passed on from one generation to the next, reinforcing thebeliefs that help for mental disorders should come from within oneself andone’s family only. Consequently persons with mental health problemswould be “Keeping it to themselves. Holding it in and believing inthe power of their friends” (Middle-aged FGD) instead of seekinghelp. Another dimension of culture that was apparent from FGDs (as well as KIIs)was the mistrust in Western medicine. Not understanding how counseling ormedicines work made one worry about approaching service providers or staying intreatment. The habit of Vietnamese people to only go see a doctor if they aresick with physical symptoms was also a hindrance to acknowledging mental illnessand seeking care for it. Challenges, including the lack of vocabulary to expressmental illness and symptoms, in the Vietnamese language, exaggerated theproblem, even among those who had some understanding of mental disorders. It wassaid in the young men FGD that: “when you classify depression as anillness, no one wants to be sick,… if you call it an illness, no onewants to have that sort of illness, and it’s not an illness that youcan physically see…” (Young men FGD). Another young mansummarized so well the influence of culture on mental illness stigma:“Us Southeast Asian, like, from my parents specifically has VietnamWar refugees. I think the reason why they don’t talk about it isbecause it’s a barrier that they have to overcome themselves, right?As refugees, as people who have been through the war…[omitted]They don’t want to believe that theyneed help, and so the trauma that they carry when they give birth to us iscarried on us as well. But due to the language barrier and also the, like, theysay with the whole health care, in Vietnam I know that they don’treally believe in Western and Eurocentric medicine. So, from their understandingof how, like from their experience with colonization or French people, and howmedicine works, they don’t believe in it” (Young men FGD).One characteristic of the Vietnamese culture that was also often mentioned byour FGD participants (as well as KIIs) was the lack of sharing and opennessbetween generations, even within a family. Grandparents, parents, and childrendo not usually share and discuss each other’s problems. Parents andgrandparents do not talk about problems because they need to appear strong andgood in front of their children; children do not talk about problems becausethey are supposed to do well in all aspects, particularly in school. Thecompetitiveness of Vietnamese and high expectations of younger generations againcome into play here and create a vicious cycle. Young people are expected to dowell in school, which put pressure on them and may result in mental healthproblems, yet, they cannot talk about it with their parents because they are notsupposed to feel bad about school, and sharing is not encouraged. The Asianmodel minority myth and the expectations of parents that their children would dowell in school and become doctors and lawyers were cited by many as a cause ofmental health problems among young people. “Our parents are refugees,they had nothing and our parents want us to achieve this AmericanDream…. [omitted] It set expectations andimages for us…. It was expected for all the Asians to be in the top10, and for, like a little quick minute I thought I wasn’t going tomake it, I was crying” (Yong men FGD). As a result, the mental healthproblems get worse. “If you’re feeling bad aboutsomething, you don’t feel like you can talk about it with anyoneelse, especially your family, because it is not something that is encouraged tobe talked about anyway, so if you are feeling poorly and you don’tfeel like you could talk to anybody, I think that just perpetuates the badfeelings” (Middle-aged women FGD). Acculturation and mental illnessstigma Acculturation, the degree of assimilation to the host society, haschanged some of the understanding of mental illness and stigmatizing attitudes.Differences across generations expressed in different FGDs indicated differencesin perceptions towards mental illness that could be attributed to acculturation.For example, the young generation understood that mental illness was a healthproblem that was prevalent but less recognized in the Vietnamese community,whereas a prominent theme among the older participants was that mental illnesswas a temporary condition due to psychological stress, that it was a conditionthat only Caucasians had. Some of the components of public stigma related tomental illness seemed to vary between generations, for example the youngestparticipants were less likely to put a label on a person with mental healthproblems, or to stereotype them, compared to the oldest and middle-agedparticipants. This was attributed to their education, exposure to the media andinformation, and to them “being more Americanized.”However, there was no evidence that acculturation played an important role inchanging the other components of public stigma, including stereotyping,separating, and status loss and discrimination. For example, the need to protectthe family reputation was so important that our young participants shared:“If you damage their image, they will disown you before you damagethat image” (Young men FGD). Young people, more likely to recognizemental health problems, were also more likely to share within the family and toseek help, but no more likely than their older counterparts to share outside ofthe family—“maybe you would go to counseling or go totherapy, but you wouldn’t tell people you’re doingthat” (Young women FGD). The youngest participants in our study werefacing a dilemma, in which they recognized mental health problems and the needfor care, yet were still reluctant to seek care or talk about it publiclybecause of fears of damaging the family reputation and not living up to theparents’ expectations. Many young participants reported that itactually made it very difficult for them to navigate mental health issuesbetween the 2 cultures, despite the awareness of the resources available.“I think it actually makes it harder. Only because you know to yourparents and the culture, and your own people, it’s taboo, andit’s something that you don’t talk about. Just knowingthat you have the resources to go seek it… You want advice from yourfamily also, but you can’t connect the appointment to your familybecause you’re afraid to express that to your parents, you know? So Ithink that plays a big part, and knowing that you are up and coming, but youdon’t want to do something to disappoint your family because they areso traditional” (Young men FGD). Some participants felt morecomfortable talking about mental health problems, like depression, if it wastheir friend who experienced it and confided in them, but they would notnecessarily felt open if it was their problem. Subtle cultural differences likethis are likely overlooked by Western service providers. One older participantsummarized it well “They [the young generation]are more Americanized. They are more open to other things[but] I think that mental health is still abarrier.” DISCUSSION/SIGNIFICANCE OF IMPACT: This studyinvestigated how different components of public stigma related to mental illnessmanifest among Vietnamese Americans, a major ethnic group in the United States,and how acculturation may influence such stigma. The findings highlightedimportant components of public stigma, including labeling and status loss, butdid not provide strong evidence of the other components within our studypopulation. Strong cultural beliefs underlined the understanding of mentalhealth and mental illness in general, and how people viewed people with mentalillness. Several findings have been highlighted in previous studies with Asianimmigrants elsewhere; for example, a study from the perspectives of health careproviders in Canada found that the unfamiliarity with Western biomedicine andspiritual beliefs and practices of immigrant women interacted with social stigmain preventing immigrants from accessing care (O’Mahony and Donnelly,2007). Fancher et al. (2010) reported similar findingsregarding stigma, traditional beliefs about medicine, and culture amongVietnamese Americans. Acculturation played a role in changing stigmatizingattitudes as evidenced in intergenerational differences. However, being moreAmericanized did not equate to being more open, having less stigmatizingattitudes, or being more willing to seek care for mental health issues.Consistent with previous studies (Pedersen and Paves, 2014), we still found somelevel of stigma among young people aged 18–35, although somecomponents were lessened with an increased level of acculturation. There wasalso a conflict among the younger generation, in which the need for mentalhealth care was recognized but accessing care was no easier for them than fortheir parent and grandparent generations. The study’s findings areuseful to adapt existing instruments to measure stigma to this population. Thefindings also have important program implications. One, they can be directlytranslated into basic supports for local primary and behavioral health careproviders. Two, they can also be used to guide and inform the development andevaluation of an intervention and an additional study to validate the findingsin other immigrant ethnic groups in the United States. Finally, based on resultsof the study, we can develop a conceptual framework that describes pathwaysthrough which social, cultural, and ecological factors can influence stigma andthe ways in which stigma acts as a barrier to accessing mental health care amongVietnamese Americans. The guiding framework then can be validated and applied infuture programs aimed to improve mental health care utilization among ethnicminorities.

Effective stakeholder engagement increases research relevance and utility. Though published principles of community-based participatory research and patient-centered outcomes research offer guidance, few resources offer effective techniques to engage stakeholders and translate their engagement into improvements in research process and outcomes. The Indiana Clinical and Translational Sciences Institute (Indiana CTSI) is home to Research Jam (RJ), an interdisciplinary team of researchers, project management professionals, and design experts, that employs human-centered design (HCD) to engage stakeholders in the research process. Establishing HCD services at the Indiana CTSI has allowed for accessible and innovative stakeholder-engaged research. RJ offers services for stakeholder-informed study design, measurement, implementation, and dissemination. RJ’s services are in demand to address research barriers pertaining to a diverse array of health topics and stakeholder groups. As a result, the RJ team has grown significantly with both institutional and extramural support. Researchers involved in RJ projects report that working with RJ helped them learn how to better engage with stakeholders in research and changed the way they approach working with stakeholders. RJ can serve as a potential model for effectively engaging stakeholders through HCD to improve translational research.

Understanding the clinical risk factors for COVID-19 disease severity and outcomes requires a combination of data from electronic health records and patient reports. To facilitate the collection of patient-reported data, as well as accelerate and standardize the collection of data about host factors, we have constructed a COVID-19 survey. This survey is freely available to the scientific community to send electronically for patients to complete online. This patient survey is designed to be comprehensive, yet not overly burdensome, to gather data useful for a range of clinical investigations, and to accommodate a wide variety of implementation settings including at a COVID-19 testing site, at home during infection or after recovery, and/or for individuals while they are hospitalized. A widely adopted standardized survey that can be implemented online with minimal resources can serve as a critical tool for combining and comparing data across studies to improve our understanding of COVID-19 disease.

OBJECTIVES/SPECIFIC AIMS: This study’s aims are to optimize the isolation and growth of chondrocytes from pig auricular cartilage; to identify the ideal seeding conditions onto 3D printed auricular bioscaffolds to maximize chondrocyte growth; and to investigate what quantity and types of host tissue can grow on the bioscaffold. Primary outcomes will include comparisons between different seeding conditions in various objective measures of bioscaffold growth and survival as listed in the methods section. Secondary outcomes will include continued optimization of bioscaffolds to minimize extrusion rates and maximize morphologic and histologic similarity to human auricular cartilage. METHODS/STUDY POPULATION: For chondrocyte-seeded scaffolds, cartilage will be collected from freshly harvested porcine auricular tissue and digested in type II collagenase. Chondrocytes derived from the harvest will be seeded into auricular PCL scaffolds using a type I collagen/hyaluronic acid composite gel, which has been previously shown to support chondrogenesis. For scaffolds containing cartilage, punch biopsies will be collected and embedded in specific areas of the scaffold previously shown to experience excessive stress/strain compared to the rest of the construct. From there, five of each chondrocyte-seeded bioscaffolds, chondrocyte-unseeded bioscaffolds, and cartilage-containing bioscaffolds will be implanted into athymic rats. Total follow up will be for six months, with outcomes as measured by clinical assessments, morphologic measurements, radiological imaging, histological analysis, biomechanical evaluation, and photodocumentation. Once these measures are obtained, we will work closely with Dr. Myra Kim, an adjunct professor with the Biostatistics Department, to appropriately analyze differences between the models. RESULTS/ANTICIPATED RESULTS: We believe that while all scaffolds (chondrocyte-seeded, chondrocyte-unseeded, and cartilage-containing) will be structurally sound, the chondrocyte-seeded scaffolds and cartilage-containing scaffolds will exhibit improved soft tissue coverage and have lower exposure and fracture rates. Additionally, between the two, we posit that there will not be appreciable differences histologically, radiologically, or morphologically. DISCUSSION/SIGNIFICANCE OF IMPACT: Auricular reconstruction is a geometrically complex and technically challenging problem. Reconstruction hinges on the physical characteristics of the deformity, patient preferences, and reconstructive materials available. The current gold standard for auricular reconstruction uses autologous rib cartilage as foundational support for overlying soft tissue and these techniques involve freehand carving of the cartilage, requiring high levels of technical skill. Harvesting the materials for this procedure is invasive, and the outcomes of the surgery are largely variable and sometimes undesirable. As alternatives, implantable scaffolds including those made from high density porous polyethylene (commercially referred to as MedPor) have been investigated. However, many of these have proven inadequate due to factors including infection, extrusion, and morphologic and biomechanical dissimilarity from native tissue. 3D printing represents an exciting new avenue through which to address many of these difficulties. Our group has previously demonstrated the successful design, production, and implantation of 3D-printed models: in auricular reconstruction, we have demonstrated the successful creation and implementation of a 3D printed ear scaffold into an athymic rodent model. We now turn our attention to optimization of seeding of our ear scaffold with chondrocytes derived from porcine auricular cartilage or with cartilage punch biopsies, all while maintaining emphasis on regulatory feasibility. With success in this arena, we will be able to provide a much less invasive and technically challenging alternative to the current gold standard, create patient-specific bioscaffolds which are more form fitting and individualized, and provide children with ear malformations better alternatives and treatments for their conditions.

OBJECTIVES/SPECIFIC AIMS: The promise and potential of connected personal health records (PHRs) has not come to fruition. This may be, in part, due to the lack of user-centered design and of a patient-centric approach to curating personal health data for use by patients. Co-design with end-users could help mitigate these issues by ensuring the software meets user’s needs, and also engages patients in informatics research. Our team partnered with patients with multiple chronic conditions to co-design a patient-centric PHR. This abstract will describe our experience with the co-design process, highlight functionalities desired by patients, and showcase the final prototype. METHODS/STUDY POPULATION: We conducted 3 design sessions (90 min per session) with patients as co-designers and employed an iterative process for software development. Patients were recruited from Chapel Hill and surrounding areas. The initial design session laid the foundation for future sessions, and began with brainstorming about what patients thought their ideal version of an engaging connected PHR would look like in terms of features and functionalities. After each software iteration, our entire design team, including our patient co-designers, was shown the prototype during a subsequent design session. Once the final prototype was developed, usability testing was conducted with patient participants. Our team then conducted a final design session to debrief about the final prototype. RESULTS/ANTICIPATED RESULTS: We started with an initial group of 12 patients (6 males) who all had diabetes and an additional comorbidity such as hypertension and hyperlipidemia. Age of participants ranged from 30 to 77 years with an average age of 56. The majority of participants were Caucasian with 1 Asian and 2 African Americans. Hemoglobin A1c values ranged from 6.0% to 9.2% with approximately half having A1c values less than the goal of 7.0%. Half the patients were aware of PHRs, majority had smartphones, and all participants had access to the Internet and used email. Two of the patients were retired engineers who had prior experience with software design. The other sessions had between 7 and 8 participants at each session, and 7 patients completed the 90-minute usability testing session. There was a core group of 7 patients who were engaged in the design and testing sessions throughout the entire 9-month study. Key features of the PHR that emerged from design sessions included the following: (1) allow for annotation of data by patients (particularly important for lab values like glucose or for physical activity); (2) calendars, to do list, and reminder functions should be linked so that an entry in one of these allows for auto-population of this data within the other sections; (3) notifications whenever new data from the electronic health record or other sources are pushed to the PHR account; (4) allow for drag and drop of photos of pills/medications taken via smartphone or from other sources so that medication list has photo of actual pills or pill bottle; (5) allow for patients to customize the order of sections in the PHR dashboard so that the sections most important to the individual patient can be displayed more prominently; (6) allow for notifications from pharmacies to be pushed to the PHR (eg, confirmation of receipt of prescription requests or alert that prescription is ready to pick up); and (7) graphical display of trends over time (patients would like to select the measures and time frames to plot for display). Patients cited the importance of data provenance so that patient-entered data Versus provider or electronic health record data could be easily differentiated. Patients also highlighted the importance of having this PHR be a “one-stop shop for all their health data” and to have meaningful data dashboards for the different types of information needed to comprehensively manage their health. Patients wished for a single PHR that could easily bring together data from multiple patient portal accounts to avoid having to manage multiple accounts and passwords. They felt that heat map displays such as those used on popular fitness tracking websites were not intuitive and that the color-coding made interpretation challenging. Participants noted that engagement in the design process made them feel that they contributed towards developing software that could not only positively impact them individually but others as well. Every patient indicated the desire to participate on future design projects. Of the 19 tasks evaluated during usability testing, only 5 tasks could not be completed (eg, adding exercise to the calendar, opening the heat map, etc.). Patients felt that the overall PHR design was clean and aesthetically pleasing. Most patients felt that the site was “pretty easy to use” (6 out of 7). The majority of participants would like to use this PHR in the future (5) and would recommend this PHR to their friends/family to use (6). DISCUSSION/SIGNIFICANCE OF IMPACT: Involving patients directly in the design process for creating a patient-centric connected PHR was essential to sustaining engagement throughout the software life cycle and to informing the design of features and functionalities desired by patients with chronic conditions.

OBJECTIVES/GOALS: Our research has three main aims: 1. Measure attitudes toward a SARS-CoV-2 vaccine among BIPOC. 2. Assess the effectiveness of race-conscious public health messages in changing attitudes toward a SARS-CoV-2 vaccine. 3. Test the efficacy of financial incentives to increase uptake of a SARS-CoV-2 vaccine. METHODS/STUDY POPULATION: We surveyed 784 unvaccinated residents of L.A. County. To recruit participants, we collaborated with Qualtrics. The survey randomized participants to one of three public health messages, as well as one of two financial compensation schemes. Twenty-five participants completed semi-structured interviews via Zoom or telephone. Interviews were audio recorded, translated into English if needed, and transcribed. The inductive, semi-structured interview guide focused on three domains: i) concerns and distrust toward a COVID vaccine, ii) policy interventions and/or government action related to a COVID vaccine, iii) non-pharmacological policy interventions related to the COVID-19 pandemic. Major emergent themes were identified and analyzed using Watkins (2012) team analysis of qualitative data steps. RESULTS/ANTICIPATED RESULTS: Many BIPOC remain vaccine hesitant: 2/3 of the survey respondents stated that they did not intend to or were not sure if they planned to get vaccinated. Follow-up interviews show that fear of vaccine side effects, bodily autonomy in choosing to get vaccinated are major concerns. However, public health measures like masking and physical distancing remain preferred safety methods for BIPOC residents. Misinformation and overcommunication in public health messaging concerning vaccine eligibility may be a major barrier to vaccine uptake among BIPOC. DISCUSSION/SIGNIFICANCE: Real world financial compensation mechanisms need to provide large enough compensation to avoid a crowding out of altruistic vaccination motivations and to effectively incentivize increased vaccine uptake. Additionally, short race-conscious public health messages were ineffective at improving vaccine attitudes.

Objectives/Goals: We aim to explore the associations of race/ethnicity and socioeconomic status (SES): 1) with grip strength, walking speed, and comorbidity index cross-sectionally and 2) with the change in comorbidity index and mortality risk over four years of follow-up in cancer survivors. Both aims will examine the potential mediating role of cytomegalovirus (CMV) infection. Methods/Study Population: This study includes 1,602 cancer survivors (mean age = 72 years, 10% Black, 54% female) from the Health and Retirement Study (HRS), a nationally representative U.S. sample followed for health outcomes until 2020. HRS measured CMV immunoglobulin G (IgG) antibody levels (from blood samples), grip strength, and walking speed in 2016. We will apply linear regression to examine the associations of race/ethnicity and SES with grip strength, walking speed, and comorbidity index cross-sectionally and with the change in comorbidity index over four years of follow-up. We will apply Cox proportional hazard regression to examine the associations of race/ethnicity and SES with mortality over four years of follow-up. In all models, we will investigate the potential mediating role of CMV infection in these associations. Results/Anticipated Results: We expect that CMV infection mediates the associations of race/ethnicity and SES with age-related health outcomes, including muscle weakness (measured by grip strength), decreased functional performance (measured by walking speed), comorbidity index, and mortality in elderly cancer survivors. Discussion/Significance of Impact: If our hypothesis is confirmed, the findings may inform physicians to closely monitor CMV infection among cancer survivors from socially disadvantaged groups and apply treatment if needed. Several oral medications for CMV exist, and CMV vaccines are currently undergoing testing in clinical trials. This will make the treatment for CMV more accessible.

OBJECTIVES/GOALS: Breast cancer has an increased requirement for lipids. The sigma-2 receptor plays a critical role in the effective uptake of lipoproteins by forming a complex with the LDL Receptor. We investigate the role of the sigma02 receptor in modulating lipid uptake pathways in breast cancers, and how this can be leveraged as a viable therapeutic strategy. METHODS/STUDY POPULATION: CRISPR/Cas9 will be used to ablate TMEM97 in the MDAMB231 and MCF7 cell lines. This study seeks to identify pathways that are dysregulated upon TMEM97 knockout (KO) by characterizing RNASeq data to identify differentially expressed genes and perform pathway analysis. RESULTS/ANTICIPATED RESULTS: Knockout of TMEM97 in breast cancer cells is is expected to decrease lipid uptake. Treatment with statins in these knockout cells is expected to result in decreased cell viability and result in a quiescent cell population. DISCUSSION/SIGNIFICANCE: This is an important mechanistic study to understand the importance of lipid homeostasis in cancer cell proliferation and how it can be targeted to improve therapeutic anti-tumor strategies. Understanding the pathways that TMEM97 modulates is vital for therapeutic strategies to curb the proliferation of breast cancer cells.

OBJECTIVES/GOALS: Analyze how the Endowment HIREC ‘s Mentoring and Career Coach Model A productive mentoring relationship is essential to advance researchers into being independent and bring extramural funds. METHODS/STUDY POPULATION: Provide Hispanic researchers mentoring and career coaching to strengthen their pathway as researcher. The HiREC’s Career Coach and Mentoring Component (CCMC) is an innovated approach to support long-lasting research mentoring relationships in our institution. This approach was developed to advance research to eliminate health disparities, promote multidisciplinary translational research in a Minority Institution and sustain research infrastructure and services, career, and workforce development initiatives. Promising Faculty are target and early and mid-career investigators interested in pursuing a research career. To implement the CCMC with the Visiting Endowed Chair a HiREC Advisory Leadership Group in Mentoring will be established, with researchers from Puerto Rico, and US mainland. RESULTS/ANTICIPATED RESULTS: Three Hispanic mid-career women from the School of Medicine and one from the School of Health Professions from the University of Puerto Rico received a HiREC Advanced Research Award of $50,000. The awardees achieved their goals; completed their research plan, research infrastructure needs, peer-reviewed publications, and submission of a competitive grant. They also provided successful perspectives on mentoring relationships in a Minority institution. Each one showed the mentor’s and mentee’s experiences as fundamental for their research advancements, productivity, leadership, and successful results. HiIREC’s mentoring component with the Visiting Endowed Chairs improves a healthy work environment and expands the research agenda for each awardee sustaining the institutional research culture. DISCUSSION/SIGNIFICANCE: A productive mentoring relationship is essential to advance researchers into being independent and bring extramural funds. Four mentees received formal, long-term guidance and endowment funds for their research infrastructure requirements with successful outcomes. HiREC contributes to building up an institutional mentoring program.

OBJECTIVES/GOALS: Health insurance status is associated with differences in access to healthcare and health outcomes. The objective of this study was to test the hypothesis that among infants born in the United States, maternal private insurance compared with public Medicaid insurance would be associated with a lower infant mortality rate (IMR). METHODS/STUDY POPULATION: This ecological study used data from the Center for Disease Control and Prevention (CDC) WONDER expanded linked birth and infant death records database 2017-2018. We included hospital-born infants from 20 to 42 weeks of gestational age (wga) if the mother had either private or Medicaid insurance. We excluded infants with congenital anomalies and infants who died due to congenital anomalies. We used negative-binomial regression adjusted for race, sex, multiple birth, and any maternal pregnancy risk factors (as defined by the CDC) to determine the difference in IMR between private and Medicaid insurance. Chi-square or Fishers exact test was used to compare differences in categorical variables between groups. RESULTS/ANTICIPATED RESULTS: We included 6,901,328 infants; 53.6% had private insurance and 46.4% were insured by Medicaid. Privately insured infants had a lower IMR compared with Medicaid insured infants (2.84/1000 vs. 5.32/1000; adjusted relative risk (aRR) 0.71; 95% confidence intervals (CI) 0.62 to 0.81; p<.0001). The privately insured had higher rates of 1st trimester prenatal care compared to those with Medicaid (85.6% vs. 66.6%; p<.00001). Rates of infant morbidity and maternal morbidity (per CDC definitions) were lower among the privately insured compared to those with Medicaid (both p<.00001). The privately insured had lower rates of preterm (9.1% vs. 11.0%), extremely preterm (0.5% vs. 0.7%), low birth weight (7.1% vs. 9.6%), and extremely low birth weight (0.5% vs. 0.7%) births compared to those with Medicaid (all p<0.001). DISCUSSION/SIGNIFICANCE: Private insurance is associated with a lower IMR compared to Medicaid insurance. Privately insured pregnancies also have higher rates of early prenatal care, less morbidity, and less preterm and low birth weight births. There may be opportunities to improve access to care and pregnancy outcomes among Medicaid insured pregnancies in the United States.

OBJECTIVES/GOALS: Patients frequently need or desire fat grafting to improve common issues such as implant visibility and contour deformity, often done as a second, staged procedure following immediate reconstruction. This study aimed to identify which patient factors and reconstructive techniques predict the need for revision with AFG after IBBR METHODS/STUDY POPULATION: Patients who underwent IBBR with either tissue expanders or implants following mastectomy from 2017 to 2021 were identified. Demographics, comorbidities, and the postoperative course were reviewed. The primary outcome variable was AFG after the initial reconstruction. Univariate and regression analyses were performed to identify factors predictive of AFG. RESULTS/ANTICIPATED RESULTS: Five-hundred twenty-nine patients were included in our analysis, with 43% having AFG. Univariate regression displayed single-stage reconstruction (OR=0.53, 95% 0.37-0.75) and previous radiation (OR 0.59, 95% 0.35-0.99) negatively predicted the need for AFG, while bilateral breast reconstruction (BBR) was a predictor (OR 2.32, 95% 1.58-3.4). On multivariate analysis, decreasing age and BBR remained predictive of AFG. The odds of AFG decreased by 3% for every one-unit increase in age (95% CI [0.96, 0.99]). Interestingly, neither pre-pectoral breast reconstruction nor specimen weight:implant ratio was associated with increased need for AFG on univariate/multivariate analysis. DISCUSSION/SIGNIFICANCE: Patients requiring AFG were likely younger and had undergone BBR with tissue expanders. Plane of implant did not appear to affect need for AFG. Knowledge of these predictive factors may help plastic surgeons in preoperative counseling before implant-based breast reconstruction.

Objectives/Goals: This study aims to understand the prevalence of trauma in young adults and what sociodemographic factors influence trauma exposure and type of trauma. It also seeks to explore functional connectivity and neural network patterns associated with trauma by analyzing resting state magnetic resonance imaging (MRI) data. Methods/Study Population: Sociodemographic data will be analyzed from participants aged 18–25 years, such as age, gender, race, ethnicity, and highest level of education completed. Trauma exposure will be assessed based on the DSM-5 criteria of trauma through phone screenings and clinical interviews. The data will be categorized based on trauma type and statistical analyses will be conducted to explore potential sociodemographic patterns related to trauma. Additionally, resting-state MRI data will be utilized to identify potential neural correlates of trauma exposure. Results/Anticipated Results: It is anticipated that sociodemographic factors such, race and ethnicity, and highest level of school completed may influence the likelihood of experiencing traumatic events. It is predicted that in the resting-state MRI analysis that there will be altered functional connectivity in trauma exposed young adults in regions such as the amygdala, hippocampus, and prefrontal cortex since those regions are implicated in emotional regulation and stress response. Some changes may also be seen in the default mode network and salience network. Discussion/Significance of Impact: Trauma exposure can alter neural circuitry, leading to emotional processing difficulties and heightened stress response, with lasting effects on mental health and brain development. Prevention efforts and targeted treatments can be guided by identification of affected brain networks and sociodemographic factors that increase trauma risk.

OBJECTIVES/SPECIFIC AIMS: To develop an algorithm that identifies post-concussion syndrome (PCS) cases and controls from among patients with mild traumatic brain injury (mTBI) in a large academic biobank. METHODS/STUDY POPULATION: The Vanderbilt University Medical Center’s (VUMC) electronic medical record (EMR) research database includes longitudinal medical record data on 2.5 million people. DNA and genotype data were also available for >225,000 of these individuals. Our algorithm used a combination of billing codes and natural language processing to apply inclusion and exclusion criteria. We defined PCS cases as those with a PCS billing code (ICD-9 310.2 or ICD-10 F07.81) and/or symptoms of PCS within 1–6 months of a qualifying mTBI. We will compare the positive predictive value of our algorithm to that of 2 simpler case selection schemes: (1) 1 instance of the PCS billing code anywhere in the medical record; and (2) 2 or more instances of the PCS billing code anywhere in the medical record. RESULTS/ANTICIPATED RESULTS: An mTBI was diagnosed in 28,720 patients regularly attending VUMC, and 528 of these patients were classified as PCS cases by our algorithm. The characteristics of our EMR sample reflected known risk factors for PCS. Our cases were more likely than controls to be female (49.4% vs. 38.4%), to have sustained a previous TBI (31.0% vs. 12.0%) and to have comorbid mood disorders. Our PCS cases were also more likely than controls to be <18 years of age (42.4% vs. 33.6%) and to have a sports-related keyword associated with the mTBI (44.1% vs. 25.2%), emphasizing the relevance of PCS to young athletes. Nonetheless, the number of PCS cases identified by our algorithm was small, and within the VUMC EMR, there were 5039 patients with 1 PCS billing code, and 2457 patients with 2 or more PCS billing codes anywhere in their EMR. Our next step is to calculate the positive predictive values of each selection scheme by manually reviewing the EMR of a selection of cases. Ultimately, we will implement the selection scheme that maximizes both positive predictive value and sample size, and in future work, we will genotype the selected patients to better understand the genetic architecture of PCS. DISCUSSION/SIGNIFICANCE OF IMPACT: EMR and biobanks are the future of human health research, and we asked whether complex algorithms or simple billing codes were best for studying the genetics of recovery after mTBI within the VUMC EMR. Our results are relevant to other studies of brain injury phenotypes within biobanks, including recovery from moderate or severe TBI, recovery from stroke, or the occurrence of delirium after routine surgery, and will help transform biobanks into fruitful research tools.

ABSTRACT IMPACT: Our work might lead to a new treatment for patients with acute myeloid leukemia OBJECTIVES/GOALS: Acute myeloid leukemia (AML) is a devastating hematologic malignancy, with dismal 5-year survival. Chimeric antigen receptor (CAR) T cells have been approved for B cell malignancies but not for AML. The goal of this study is to explore the safety and efficacy of CAR T cells targeting CD105 (endoglin) to treat AML. METHODS/STUDY POPULATION: We have constructed human and murine CAR T cells targeting CD105. The CARs were created by sequencing the V(D)J regions of hybridomas and designing single chain variable fragments that target CD105 which were subsequently introduced in a CAR backbone via Gibson assembly. The CAR T cells were produced via transduction using retrovirus or lentivirus. Leukemia cell lines were assessed for CD105 expression with flow cytometry. Killing assays were performed via measurement of luminescence of target cells after co-culture with CAR T cells. Activation assays were performed with co-culture of CAR T cells and target cells and measurement of activation markers with flow cytometry. To assess in vivo efficacy and safety, murine CAR T cells were infused into C57BL/6J mice carrying B16 melanoma after lymphodepletion. RESULTS/ANTICIPATED RESULTS: All human leukemia cell lines assessed (Nalm6, MOLM-14, MV4-11, Kasumi-1, THP-1) expressed some degree of endoglin apart from the T cell leukemia Jurkat. Human CD105 CAR T cells were activated by co-culture with leukemia cell lines and effectively killed leukemia cells in vitro in a CD105-specific manner. Murine CAR T cells killed efficiently both murine solid tumors (B16 melanoma) and murine leukemias (C1498) in vitro. Murine CAR T cells did not exhibit any toxicity when infused after low-dose lymphodepletion (cyclophosphamide 100mg/kg) but caused significant morbidity after higher doses (cyclophosphamide 200mg/kg). Murine CAR T cells delayed the growth of B16 melanoma in immunocompetent mice. DISCUSSION/SIGNIFICANCE OF FINDINGS: We have constructed human and murine CD105 CAR T cells with excellent activity in vitro. The activity of human CD105 CAR T cells in xenografts and the biologic relevance of the toxicity of murine CD105 CAR T cells in humans needs to be further investigated. CD105 CAR T cells might prove an important therapeutic option for patients with AML.

This abstract is based on unpublished data. OBJECTIVES/GOALS: The estimates of unbiased first-degree relatives (FDRs) risk of cancers would enhance genetic counseling of at-risk FDRs in families where the pancreatic cancer (PC) proband carrying a germline variant. This study aims at quantifying gene-specific risks of six cancers among FDRs of PC patients with germline variants in cancer-associated genes. METHODS/STUDY POPULATION: In the prospective, clinic-based Mayo Clinic Biospecimen Resource for Pancreas Research registry, 4,562 PC patients had previously undergone germline genetic testing for pancreatic cancer-associated genes through either research studies or clinical testing. Of these, 234 PC probands were found to carry germline pathogenic/likely pathogenic variants (PLPV) among 9 genes of interest and had provided detailed demographic and cancer data on their FDRs by questionnaire. We focused on six cancer types (ovary, breast, uterus, pancreas, colon, and malignant melanoma) in FDRs as reported by the probands. Standardized incidence ratios were calculated to estimate risk of six cancers among FDRs of PC patients carrying PLPV by gene. RESULTS/ANTICIPATED RESULTS: 1,670 FDRs (mean age 58.1+17.8SD; 48.9% female) were included in the study. We found significantly increased risk of ovarian cancer in female FDRs of PC probands who carry PLPV in BRCA1 (SIR 9.49, 95%CI:3.06-22.14) or BRCA2 (3.72, 95%CI:1.36-8.11), and breast cancer risks were higher with BRCA2 (2.62, 95%CI:1.89-3.54). Uterine cancer risk was increased in FDRs of PC probands who carry PLPV for Lynch Syndrome mismatch repair (MMR) (6.53, 95%CI:2.81-12.86). PC risk was also increased (ATM 4.53, 95% CI:2.69-7.16; BRCA2 3.45, 95%CI:1.72-6.17; CDKN2A 7.38, 95%CI:3.18-14.54; PALB2 5.39, 95%CI:1.45-13.79). Increased colon cancer risk was observed in FDRs of probands who carried MMR PLPV (5.83, 95%CI:3.70-8.75), while melanoma risk was elevated for FDRs of probands with CDKN2A PLPV (7.47, 95%CI:3.97-12.77). DISCUSSION/SIGNIFICANCE: PLPV in nine syndrome-associated genes in PC probands are associated with increased risk of six cancers in FDRs. The findings underscore the importance of risk estimation of various other cancers in PC families for screening, early detection, intervention, and cascade genetic testing.

OBJECTIVES/SPECIFIC AIMS: This study aims to estimate patient attitudes and receptiveness towards stem cell-based cell replacement devices to management Type 1 Diabetes. The primary outcomes of this study are mean response values to questions interrogating patient attitudes, knowledge, and receptiveness. METHODS/STUDY POPULATION: A RedCap survey was generated for this study. 100 participants will be drawn from Mayo Clinic Rochester, MN patients living with Type 1 Diabetes. RESULTS/ANTICIPATED RESULTS: Response values will be used to estimate broader patient attitudes. DISCUSSION/SIGNIFICANCE OF IMPACT: The response values of this survey will help inform future directions of cell replacement device development. Additionally, understanding patient attitudes may be useful in crafting more effective education strategies.