OBJECTIVES/GOALS: To characterize the oncogenic potential of HNSCC cell lines harboring 17 non-canonical PIK3CA mutations. METHODS/STUDY POPULATION: Non-canonical PIK3CA mutant constructs generated via site-directed mutagenesis are subcloned into doxycycline-inducible vector pLVX-Puro. Serum-dependent HNSCC cell line (PCI-52-SD1) is then stably transfected with vectors and undergo doxycycline-induction. Cell survival is determined by depriving cells of fetal bovine serum for 72 hours and quantifying remaining cells with 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Cell proliferation and migration is evaluated with colony formation assays and transwell assays respectively. RESULTS/ANTICIPATED RESULTS: To date, the survival behavior of eight non-canonical mutants was assessed. Three mutants – Q75E, V71I, and E970K – exhibited 18.7-26.7% greater survival rate relative to cells transfected with wild-type. Five mutants – R519G, Y606C, W328S, C905S, and M1040I – demonstrated survival rates that differed only by −4.3% to +6.6% relative to wild-type. We hypothesize the three activating mutants that exhibited increased survival will also demonstrate increased cell proliferation and migratory behavior whereas the three neutral mutants will not differ from control. DISCUSSION/SIGNIFICANCE OF IMPACT: Ongoing HNSCC PI3K inhibitor trials could be more effective if all PIK3CA hyperactivation mutations are known. Identifying non-canonical mutation effects could result in greater efficacy if drugs are restricted only to those with activating mutations. CONFLICT OF INTEREST DESCRIPTION: JRG and DEJ are co-inventors of cyclic STAT3 decoy and have financial interests in STAT3 Therapeutics, Inc. STAT3 Therapeutics, Inc. holds an interest in a cyclic STAT3 decoy oligonucleotide. The remaining authors declare no conflicts.

OBJECTIVES/GOALS: Learners will:

• Identify social structures that serve as root causes of health disparities

• Critically evaluate the ways in which racism, culture, and power perpetuate disparity

• Use critical reflection to shape their research and advocate for institutional change

METHODS/STUDY POPULATION: The Integrated Translational Health Research Institute of Virginia (iTHRIV) Health Equity curriculum provides a lens for participants to view health disparities, social structures that create and perpetuate disparities, and the path to a more equitable future. This longitudinal workforce curriculum incorporates the principles of critical race theory (CRT), including: race as a social construct, structural determinism, intersectionality, and the social construction of knowledge. Learners gain practical experience through facilitated group discussions and critical reflection of their own work including research question design, recruitment, dissemination, and enhancing the faculty pipeline. RESULTS/ANTICIPATED RESULTS: To measure the impact of the curriculum, we will evaluate learners’ participation in mentoring activities for persons from underrepresented backgrounds; participation in local and national diversity and inclusion efforts; engagement in community-based research; ability to account for implicit bias and power imbalances in their research design, including in recruitment and retention; and share research findings with community members and research participants. Evaluation strategies will include quantitative and qualitative methodologies. DISCUSSION/SIGNIFICANCE OF IMPACT: There is growing recognition of the impact of racism on the development and perpetuation of health disparities. Public health critical race praxis (an adaptation of CRT) is emerging as a theoretical framework to empower researchers to challenge the status quo in order to achieve health equity.

We examined whether change in added sugar intake is associated with change in δ13C, a novel sugar biomarker, in thirty-nine children aged 5–10 years selected from a Colorado (USA) prospective cohort of children at increased risk for type 1 diabetes. Reported added sugar intake via FFQ and δ13C in erythrocytes were measured at two time points a median of 2 years apart. Change in added sugar intake was associated with change in the δ13C biomarker, where for every 1-g increase in added sugar intake between the two time points, there was an increase in δ13C of 0⋅0082 (P = 0⋅0053), independent of change in HbA1c and δ15N. The δ13C biomarker may be used as a measure of compliance in an intervention study of children under the age of 10 years who are at increased risk for type 1 diabetes, in which the goal was to reduce dietary sugar intake.

The National Institutes of Health requires data and safety monitoring boards (DSMBs) for all phase III clinical trials. The National Heart, Lung and Blood Institute requires DSMBs for all clinical trials involving more than one site and those involving cooperative agreements and contracts. These policies have resulted in the establishment of DSMBs for many implementation trials, with little consideration regarding the appropriateness of DSMBs and/or key adaptations needed by DSMBs to monitor data quality and participant safety. In this perspective, we review the unique features of implementation trials and reflect on key questions regarding the justification for DSMBs and their potential role and monitoring targets within implementation trials.

The objective of this study is to compare aneuploidy rates between three distinct areas of the human trophectoderm: mural, polar and a region in between these two locations termed the ‘mid’ trophectoderm. This is a cohort study on in vitro fertilization (IVF) patients undergoing comprehensive chromosome screening at the blastocyst stage at a private IVF clinic. All embryos underwent assisted hatching on day 3 with blastocyst biopsy and comprehensive chromosome screening. Biopsied blastocysts were divided into three groups depending on which area (polar, mid, or mural) of the trophectoderm was protruding from the zona pellucida and biopsied. Aneuploidy rates were significantly higher with cells from the polar region of the trophectoderm (56.2%) compared with cells removed from the mural region of the trophectoderm (30.0%; P = 0.0243). A comparison of all three areas combined also showed a decreasing trend, but this did not reach clinical significance, polar (56.2%), mid (47.4%) and mural trophectoderm (30.0%; P = 0.1859). The non-concordance demonstrated between polar and mural trophectoderm can be attributed to biological occurrences including chromosomal mosaicism or procedural differences between embryologists.

OBJECTIVES/SPECIFIC AIMS: To evaluate the ability of various techniques to track changes in body fluid volumes before and after a rapid infusion of saline. METHODS/STUDY POPULATION: Eight healthy participants (5M; 3F) completed baseline measurements of 1) total body water using ethanol dilution and bioelectrical impedance analysis (BIA) and 2) blood volume, plasma volume and red blood cell (RBC) volume using carbon monoxide rebreathe technique and I-131 albumin dilution. Subsequently, 30mL saline/kg body weight was administered intravenously over 20 minutes after which BIA and ethanol dilution were repeated. RESULTS/ANTICIPATED RESULTS: On average, 2.29±0.35 L saline was infused with an average increase in net fluid input-output (I/O) of 1.56±0.29 L. BIA underestimated measured I/O by −3.4±7.9%, while ethanol dilution did not demonstrate a measurable change in total body water. Carbon monoxide rebreathe differed from I-131 albumin dilution measurements of blood, plasma and RBC volumes by +0.6±2.8%, −5.4±3.6%, and +11.0±4.7%, respectively. DISCUSSION/SIGNIFICANCE OF IMPACT: BIA is capable of tracking modest changes in total body water. Carbon monoxide rebreathe appears to be a viable alternative for the I-131 albumin dilution technique to determine blood volume. Together, these two techniques may be useful in monitoring fluid status in patients with impaired fluid regulation.

Objectives: Apathy is a debilitating symptom of Huntington’s disease (HD) and manifests before motor diagnosis, making it an excellent therapeutic target in the preclinical phase of Huntington’s disease (prHD). HD is a neurological genetic disorder characterized by cognitive and motor impairment, and psychiatric abnormalities. Apathy is not well characterized within the prHD. In previous literature, damage to the caudate and putamen has been correlated with increased apathy in other neurodegenerative and movement disorders. The objective of this study was to determine whether apathy severity in individuals with prHD is related to striatum volumes and cognitive control. We hypothesized that, within prHD individuals, striatum volumes and cognitive control scores would be related to apathy. Methods: We constructed linear mixed models to analyze striatum volumes and cognitive control, a composite measure that includes tasks assessing with apathy scores from 797 prHD participants. The outcome variable for each model was apathy, and the independent variables for the four separate models were caudate volume, putamen volume, cognitive control score, and motor symptom score. We also included depression as a covariate to ensure that our results were not solely related to mood. Results: Caudate and putamen volumes, as well as measures of cognitive control, were significantly related to apathy scores even after controlling for depression. Conclusions: The behavioral apathy expressed by these individuals was related to regions of the brain commonly associated with isolated apathy, and not a direct result of mood symptoms. (JINS, 2019, 25, 462–469)

This article examines historic gendered negotiations over identity, descent, and access to land along Uganda’s southern littorals by thinking about belonging with an exceptionally diverse assemblage of small fish. Based on accounts from contemporary littoral residents, the comparative historic ethnographic record, and fisheries scientists, the article describes the reproductive practices of enkejje as they may have been visible from Uganda’s southern littorals and situates the practices of raising socially recognized children within broader historical developments of clanship and public healing. Attending to these small fish in moments and in places where groups are under construction reorients analyses of descent politics beyond an often implicit focus on male interests towards the work of grandmothers, aunts, and mothers in debating the qualities of the most foundational units of the body politic – male and female children constituted relationally as persons through their status as legitimate family members.

Literature on ethnicity in Africa meets literature on multispecies ethnography to their mutual benefit. Multispecies ethnography considers people together with other-than-human beings, insisting the figure of the human is an interspecific one. We explore the ways in which multispecies ethnography needs history as part of a story about power and politics. But, the burden of the essay argues that historians of ethnicity need multispecies ethnographers’ embrace of a broader canvas of life, in motion at many scales. Historians of ethnicity need a greater awareness of change and continuity in the presence of other-than-human life forms, over time. Those same historians also might adopt the readiness of multispecies ethnographers to recognize other than the descent metaphor at the heart of thinking and making groups.

OBJECTIVES/SPECIFIC AIMS: To respond to the need for a simple tool to answer individual researchers questions: Exactly what training do I need to complete for my study and my role? Where can we go to find a comprehensive record of my research training? METHODS/STUDY POPULATION: Identify the factors that determine what training is required for each role (i.e., PI, coordinator, biostatistician) at the University, their role on the research study, type of funding, population being studied and responsibilities/duties on the research team. Develop an inventory of training required according to federal and local regulations and guidelines. Identify other related factors that ensure ongoing compliance for research professionals (i.e., medical licenses, CVs, immunizations, and credentials). Collaborate with programming professionals to explore and confirm the feasibility of such a Web site. Incorporate formal usability and pilot testing as part of the programming design process. Develop User Guide and Marketing and Launch plan for users and supervisors. Implement phased launch of the site with Google analytics, and evaluate the experience of phase I users. RESULTS/ANTICIPATED RESULTS: Three months user data and evaluation results demonstrated: 149 users created Training Roadmaps on the site. Users were from 67 different department codes, with the Department of Psychiatry the primary user. 20 users responded to a survey three months after launch. Research coordinators were the primary focus for phase I and represented almost half of the users. Survey respondents rated the site ease of use and clarity of the site as its greatest benefit. DISCUSSION/SIGNIFICANCE OF IMPACT: In September 2017, CTSI launched a new web-based training tool exclusively for University of Minnesota clinical research professionals who work with human participants, and their supervisors. The Human Research Training Web site is a free, easy-to-use tool to help identify and maintain the appropriate training, certification, credentials, and immunizations needed to perform University of Minnesota research with human participants. The Web site offers the University’s first systematic way to identify which research training is necessary for each research professional, and a system to track and maintain training compliance. Training records and information from the University of Minnesota’s central databases are securely integrated into this tool. Our Web site tool enhances research compliance. Any given study team member’s training requirements vary based on several criteria such as: role at the University, role on the research study, type of funding, population being studied and responsibilities/duties on the research study. The research training Web site generates required and optional training based on individuals’ responses to these questions. This Web site also links to the training, which decreases error in taking the wrong training. Furthermore, it provides completion data for research training and is a repository for vital study information such as: medical licenses, CVs, and credentials. Supervisors are able to view training and credentials. They are alerted when one of their employee’s licenses or certificates are about to expire. Uses-to-date and evaluation feedback have informed the need for a second phase of Web site enhancements. This site will reside in both the CTSI Web site and the HRPP Web site. A link will be sent to all new University research employees upon hiring. The Human Research Training Web site will likely have applicability to other universities in addition to the University of Minnesota.

OBJECTIVES/SPECIFIC AIMS: To increase knowledge and application of clinical research coordinator competencies among Research Professionals at the University of Minnesota. METHODS/STUDY POPULATION: The UMN’s CTSI developed and piloted a Foundations for Research Professionals training program comprised of: a baseline assessment, 7 online modules, 4 in-person training sessions, video and reading assignments and a post assessment, which totaled 30–35 hours of training and covered the following topics: preparing for a study, study management, participant recruitment and engagement, assessing capacity to consent and the informed consent process. This course also provides valuable resources and connections to online references and materials. The competencies for this program were based on work of the Joint Task Force for Clinical Trial Competency. RESULTS/ANTICIPATED RESULTS: 30 clinical research professionals completed the pilot program and averaged an increase of 6.5% from baseline assessment to post assessment. Participants were asked to rate their confidence on a variety of role-based competencies at the time of preassessments and postassessments. Trends show an increase in confidence for all competency areas after completion of the training program. DISCUSSION/SIGNIFICANCE OF IMPACT: Developing a workforce of competent research professionals is integral to improve the efficiency, quality, and ethics of research. The Foundations for Research Professionals training program increased knowledge of clinical research coordinator competencies. We will assess impact on application of the competencies 6 months after completion of the program. Our next steps include offering the training program as a 2-week session on an ongoing basis for new coordinators at the University of Minnesota.

Objectives: The present study examined differences in neurocognitive outcomes among non-Hispanic Black and White stroke survivors using the NIH Toolbox-Cognition Battery (NIHTB-CB), and investigated the roles of healthcare variables in explaining racial differences in neurocognitive outcomes post-stroke. Methods: One-hundred seventy adults (91 Black; 79 White), who participated in a multisite study were included (age: M=56.4; SD=12.6; education: M=13.7; SD=2.5; 50% male; years post-stroke: 1–18; stroke type: 72% ischemic, 28% hemorrhagic). Neurocognitive function was assessed with the NIHTB-CB, using demographically corrected norms. Participants completed measures of socio-demographic characteristics, health literacy, and healthcare use and access. Stroke severity was assessed with the Modified Rankin Scale. Results: An independent samples t test indicated Blacks showed more neurocognitive impairment (NIHTB-CB Fluid Composite T-score: M=37.63; SD=11.67) than Whites (Fluid T-score: M=42.59, SD=11.54; p=.006). This difference remained significant after adjusting for reading level (NIHTB-CB Oral Reading), and when stratified by stroke severity. Blacks also scored lower on health literacy, reported differences in insurance type, and reported decreased confidence in the doctors treating them. Multivariable models adjusting for reading level and injury severity showed that health literacy and insurance type were statistically significant predictors of the Fluid cognitive composite (p<.001 and p=.02, respectively) and significantly mediated racial differences on neurocognitive impairment. Conclusions: We replicated prior work showing that Blacks are at increased risk for poorer neurocognitive outcomes post-stroke than Whites. Health literacy and insurance type might be important modifiable factors influencing these differences. (JINS, 2017, 23, 640–652)

The xenon plasma focused ion beam instrument (PFIB), holds significant promise in expanding the applications of focused ion beams in new technology thrust areas. In this paper, we have explored the operational characteristics of a Tescan FERA3 XMH PFIB instrument with the aim of meeting current and future challenges in the semiconductor industry. A two part approach, with the first part aimed at optimizing the ion column and the second optimizing specimen preparation, has been undertaken. Detailed studies characterizing the ion column, optimizing for high-current/high mill rate activities, have been described to support a better understanding of the PFIB. In addition, a novel single-crystal sacrificial mask method has been developed and implemented for use in the PFIB. Using this combined approach, we have achieved high-quality images with minimal artifacts, while retaining the shorter throughput times of the PFIB. Although the work presented in this paper has been performed on a specific instrument, the authors hope that these studies will provide general insight to direct further improvement of PFIB design and applications.