OBJECTIVES/SPECIFIC AIMS: Our objective was to develop and evaluate a machine learning pipeline that uses electronic health record (EHR) data to predict acute kidney injury (AKI) during rehospitalization for patients who did not have an AKI episode in their most recent hospitalization. METHODS/STUDY POPULATION: The protocol under which this study falls was given exempt status by our institutional review board. The fully deidentified data set, containing all adult hospital admissions during a 2-year period, is a combination of administrative, laboratory, and pharmaceutical information. The administrative data set includes International Classification of Diseases, 9th Revision (ICD-9) diagnosis and procedure codes, Current Procedural Terminology, 4th Edition (CPT-4) procedure codes, diagnosis-related grouping (DRG) codes, locations visited in the hospital, discharge disposition, insurance, marital status, gender, age, ethnicity, and total length of stay. The laboratory data set includes bicarbonate, chloride, calcium, anion gap, phosphate, glomerular filtration rate, creatinine, urea nitrogen, albumin, total protein, liver function enzymes, and hemoglobin A1c. The pharmacy data set includes, for each medication, a description, pharmacologic class and subclass, and therapeutic class. Data preprocessing was performed using Python library Pandas (McKinney, 2011). Top-level binary representation (Singh, 2015) was used for diagnosis and procedure codes. Categorical variables were transformed via 1-hot encoding. Previous admissions were collapsed using rules informed by domain expertise (eg, the most recent age or sum of assigned diagnosis codes were retained as elements in the feature vector). We excluded any patient without at least 1 rehospitalization during the time window. We excluded any admission with or without AKI where AKI was also present in the most recent hospitalization. For comparison, we do not exclude such admissions for an identical experiment in which we considered any AKI event as a positive sample (regardless of AKI presence in the most recent hospitalization). We defined an AKI event as an assignment of any of the acute kidney failure (AKF) ICD-9 codes [584.5, AKF with lesion of tubular necrosis, 584.6, AKF with lesion of renal cortical necrosis, 584.7, AKF with lesion of renal medullary (papillary) necrosis, 584.8, AKF with other specified pathological lesion in kidney, or 584.9, AKF, unspecified]. Since diagnosis codes are believed to be specific but not sensitive for AKI (Waikar, 2006), we supplemented them using creatinine for patients who had laboratory values. Diagnosis was made according to the Kidney Disease: Improving Global Outcomes (KDIGO) Practice Guidelines (AKI defined as a 1.5-fold or greater increase in serum creatinine from baseline within 7 d or 0.3 mg/dL or greater increase in serum creatinine within 48 h). We report preliminary model discrimination via area under the receiver operating characteristic curve (AUC) using k-fold cross validation grouped by patient identifier (to ensure that admissions from the same patient would not appear in the training and validation set). It was confirmed that the prevalence of positive samples in the entire data set was maintained in each fold. Python library Sci-kit Learn (Pedregosa, 2011) was used for pipeline development, which consisted of imputation, scaling, and hyper-parameter tuning for penalized (l1 and l2 norm) logistic regression, random forest, and multilayer perceptron classifiers. All experiments were stored in IPython (Pérez, 2007) notebooks for easy viewing and result reproduction. RESULTS/ANTICIPATED RESULTS: There were 107,036 adult patients that accounted for 199,545 admissions during a 2-year window. Per admission, there were at most 54 ICD-9 diagnoses, 38 ICD-9 procedures, 314 CPT-4 procedures, and 25 hospital locations visited. The admissions were 55% female, the average age was 46±standard deviation 20, and average length of stay was 2.5±8.0 days. We excluded 2360 admissions that involved an AKI event that directly followed an admission with an AKI event and 4130 admissions that did not involve an AKI event but directly followed an admission with an AKI event. In total, there were 4561 (5.3%) positive samples (AKI during rehospitalization without AKI in the previous stay) generated by 3699 unique patients and 81,458 negative samples (non-AKI during rehospitalization without AKI in the previous stay) generated by 31,831 unique patients. When using any AKI event as a positive sample (regardless of whether or not AKI was in the most recent stay), the prevalence was 7.3% (6921 positive samples generated by 4395 unique patients and 85,588 negative samples generated by 33,287 unique patients). Best results were achieved with a code precision of 3 digits for which we had a total of 4556 features per patient. Fitted hyper-parameters corresponding to each classifier were logistic regression with l1 penalty C as 2×10−3; logistic regression with l2 penalty C as 1×10−6; random forest number of estimators as 100, maximum depth as 3, minimum samples per leaf as 50, minimum samples per split as 10, and entropy as the splitting criterion; and multilayer perceptron l2 regularization parameter α as 15, architecture as 1 hidden layer with 5 units, and learning rate as 0.001. Five-fold stratified cross validation on the development set yielded AUC for logistic regression with l1 penalty average 0.830±0.006, logistic regression with l2 penalty 0.796±0.007, random forest 0.828±0.007, and multilayer perceptron 0.841±0.005. In an identical experiment for which an AKI event was considered a positive sample regardless of AKI presence in the most recent stay, we had 4592 features per sample with the same code precision. Five-fold stratified cross validation on the development set with identical settings for the hyper-parameters yielded AUC for logistic regression with l1 penalty average 0.850±0.004, logistic regression with l2 penalty 0.819±0.006, random forest 0.853±0.004, and multilayer perceptron 0.853±0.006. DISCUSSION/SIGNIFICANCE OF IMPACT: Our objective was to investigate the feasibility of using machine learning methods on EHR data to provide a personalized risk assessment for “unexpected” AKI in rehospitalized patients. Preliminary model discrimination was good, suggesting that this approach is feasible. Such a model could aid clinicians to recognize AKI risk in unsuspicious patients. The authors recognize several limitations. Since our data set corresponds to a time-window sample, patients with high frequency of hospital utilization are likely overrepresented. Similarly, our data set contains records from only 1 hospital network. Although we supplement with laboratory-based diagnosis, using diagnosis codes as labels is problematic as numerous reports suggest low sensitivity of codes for AKI. Future work includes calibration analysis, incremental updating (“online learning”), and a representation learning-based (“deep learning”) extension of the model.

OBJECTIVES/SPECIFIC AIMS: Earlier pubertal timing has been associated with risk for depression, particularly in girls (e.g., Keenan etal., 2014). Evidence suggests pubertal timing in girls also relates to alterations in the microstructural properties of brain white matter tracts in late adolescence (Chahal etal., 2018), and structural connectivity of cingulate and frontal regions (Chahal etal., in prep), though differences in pubertal development in both boys and girls have not been examined in the context of brain functional connectivity (FC). Individual differences in the course of puberty may have enduring effects on functional coupling among brain regions that may contribute to the risk for psychopathology. To address this question, we explored the relation between pubertal timing and tempo with depression symptoms (age 16). Then, we examined whether brain network FC (age 16) associates with pubertal indices and predicts concurrent and later depressive symptoms (age 18). METHODS/STUDY POPULATION: Sixty-eight adolescents (37 females) completed the Mini-Mood and Anxiety Symptom Questionnaire (MASQ; Clark & Watson, 1995) at ages 14-18. Gompertz growth curve modelling of pubertal development (age 10-15; Waves 1-6) was used to estimate pubertal timing and tempo per individual, separately for males and females (e.g., Chahal etal., 2018). Resting-state MRI data (age 16) were parcellated into 264 cortical and subcortical regions to create region-to-region FC matrices based on correlations of time-series. Individual matrices were fed to the GraphVar program (Kruschwitz etal., 2015) to assess the interaction of pubertal timing and pubertal tempo with functional network connectivity using Network-based statistic (NBS; Zalesky etal., 2010). Subnetworks showing alterations in relation to pubertal timing and tempo were then examined in association with concurrent (age 16) symptoms and used to predict future depressive symptoms (age 18). RESULTS/ANTICIPATED RESULTS: In all youth, earlier pubertal timing was associated with higher depressive symptoms at age 16 (p<.018). This association was stronger in girls with slower pubertal tempo (p<.039). Interregional connectivity analyses revealed that the interaction of earlier pubertal timing and slower tempo was associated with lower FC between the left cingulate gyrus and right precuneus (p<.0001), regions implicated in emotion processing (i.e., Affective Processing Network) and self-referential thinking (i.e., Default Mode Network). FC of the three other emotion- and self-referential processing network regions (i.g., left insula, superior parietal lobule, and precuneus) was lower in youth with greater age 16 depressive symptoms (p<.0001). Finally, lower FC of of the left and right inferior parietal lobule predicted greater depressive symptoms at age 18 (p<.0001). In summary, FC of overlapping affective and default mode network areas was related to earlier pubertal timing and higher concurrent and future depressive symptoms. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings demonstrate individual differences in pubertal maturation are associated with depressive symptoms and differences in brain connectivity in mid-adolescence. Early pubertal development was associated with greater depression symptoms and lower FC of brain regions involved in emotion regulation and self-referential processing. Further, FC between these regions predicted higher depression symptoms two years later. These neurobiological mechanisms may, in part, underlie the link between off-time pubertal development and the risk for depression. These findings also have important implications for precision psychiatry, as we show that a risk-factor of depression (early pubertal timing) may manifest in developing neurobiology in region-specific ways. Previous network models of depression (e.g., Li etal., 2018) implicated affective network connectivity in sustained negative mood and the default mode/ self-referential network in rumination. Other networks implicated in these past models include the reward network, which may be involved in anhedonia and loss of pleasure. Our study only found associations between affective and self-referential regional connectivity, pubertal maturation, and depression, suggesting that pubertal risk factors may relate more closely with emotion-regulation and self-referential processing deficits.

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/Goals: To evaluate the translational reliability, reproducibility, diagnostic performance, and subgroup equity of multimodal artificial intelligence (AI) models for dermatology triage across multiple model platforms. Methods/Study Population: Limited access to dermatology expertise delays diagnosis and care, motivating development of multimodal AI systems that integrate clinical images with patient data for triage. We assembled 200 biopsy-confirmed PAD-UFES-20 lesions (melanoma, keratinocyte carcinoma, benign) with paired images and metadata, prioritizing demographic balance. Six multimodal AI models (GPT-5, GPT-5-mini; Gemini 2.5 Pro, Gemini 2.5 Flash; Claude Sonnet-4, Claude Opus-4) analyzed these lesions with identical prompts predicting diagnostic probabilities, triage (urgent vs routine), and rationale. Outcomes included sensitivity, specificity, AUROC, F1, and subgroup equity. Model rationales were reviewed for interpretability, and subset re-prompting tested reproducibility for translational robustness. Results/Anticipated Results: Across six models, sensitivity range was 0.89–1.00, specificity 0.21–0.65, AUROC 0.77–0.87, and F1 scores 0.72–0.81. GPT-5 achieved the most balanced performance (0.92 sensitivity, 0.65 specificity, AUROC 0.87, F1 0.81), while Gemini 2.5 Pro and Flash reached perfect sensitivity but low specificity (0.21–0.25). Claude Sonnet-4 showed near-perfect sensitivity (0.99) but over-called benign cases (0.24 specificity), while Opus-4 had the lowest sensitivity (0.89). Urgent triage aligned with dermatologist biopsy patterns (87–97%), and sensitivity was consistent across sex and skin type (p ≥ 0.29). Subset re-prompting produced similar results, supporting reproducibility. Model rationales reflected dermatologic reasoning, supporting interpretability, and translational readiness. Discussion/Significance of Impact: Multimodal AI models showed balanced diagnostic performance for dermatology triage, with platform-specific trade-offs between sensitivity and specificity. Subgroup equity, interpretable rationales, and subset reproducibility define key elements for reliable translation into dermatology workflows and prospective validation.

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: 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: 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: 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/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.