OBJECTIVES/SPECIFIC AIMS: The specific objectives of this project are (1) identify, test, and validate the parameters for a simplified NLOM imaging probe that will provide specific research and point-of-care information on biofilm presence, therapeutic need and response of individual wounds to treatment. (2) Identify specific proteomic and metabolomic biomarkers of (i) wound susceptibility to infection, (ii) wound response to the most commonly used antibacterial measures in wounds, and (iii) establish criteria for more effective interventions. METHODS/STUDY POPULATION: First, optimal use parameters for NLOM including illumination, field of view, focal length, linear Versus concentric image acquisition, detection and filter wavelengths were identified. Parameters for evaluation included ease and speed of imaging, ability to map diagnostic criteria. Next, using the optimised NLOM imaging modality in bacterial biofilm isolates and subsequently a rabbit ear model of biofilm wound infection, proteomic and metabolomic biomarkers of susceptibility to infection were identified. The effects of 2 standard debridement and anti-infective treatments, polyvidone-iodine solution or cetrimide 15%+ chlorhexidine gluconate 1.5% were mapped in situ for up to 10 days using the NLOM probe. RESULTS/ANTICIPATED RESULTS: Using the novel custom NLOM probe, high resolution mapping of wound biofilm infection, as well as the underlying tissue was performed throughout the onset, development, treatment, and resolution of wound biofilm infection. Specific microbiological, microstructural, oxygenation, and pH parameters were mapped at defined surface and subsurface locations and time-points. Findings included the determination that some standard antimicrobial formulations provide a supportive environment for wound infection, and that micro-channels within the biofilm and their interface with the tissues serve as an important predictor and indicator of wound infection establishment, progression, and response. DISCUSSION/SIGNIFICANCE OF IMPACT: The novel multimodality in vivo NLOM imaging approach establishes an important tool for earlier and more specific diagnosis of wound infection risk, virulence, and invasiveness along with markers of successful treatment, and a simple clinical imaging tool for improving wound infection prevention and treatment.

OBJECTIVES/SPECIFIC AIMS: The goal of this project is to determine the role of ETV6 in early B-cell development and define how germline ETV6 mutations result in predisposition to leukemia. METHODS/STUDY POPULATION: Gene expression commons were queried for expression levels of Etv6 and Pax5 at different stages of hematopoiesis. Mouse bone marrow was isolated and fractioned into cells committed to the B cell lineage via B220+ and CD43+ staining by flow cytometry and then separated into the following fractions: Fraction A (CD24low, CD19−), Fraction B (CD19+, CD24+, BP1−), and Fraction C (CD19+ CD24+ BP1+). Wild-type or germline mutant P214L ETV6 were cloned in an MiG vector and expressed in Ba/F3 cells. ChIP-PCR was performed by cross-linking proteins to DNA with 1% formaldehyde for 10 minute at room temperature, followed by cell lysis with RIPA buffer. Lysates were sonicated to shear DNA to a length of 200–1000 base pairs, then Protein A agarose beads were used to clean and immunoprecipitate chromatin. RESULTS/ANTICIPATED RESULTS: We observed that Etv6 is highly expressed in hematopoietic stem and lymphoid progenitor cells through the pre-pro-B stage (FrA), but its expression is significantly reduced in fraction B and thereafter (p<0.0001). Etv6 expression decreases as B cells develop and is negatively correlated with Pax5 expression (r2=0.9993; p=0.0167). We next confirmed the expression patterns of ETV6 and PAX5 during B cell development in human samples. We found that ETV6 expression was higher in the early B cell fraction (CD10+, CD34+, CD19−, and CD20−) compared to the pre-B cell fraction (CD10+, CD34−, CD19+, CD20−). Conversely, we observed that PAX5 expression was higher in the preB cell fraction compared with the early B cell fraction. In Ba/F3 cells expressing ETV6 constructs, ETV6, but not ETV6 P214L overexpression significantly decreased Pax5 expression (p≤0.05). ETV6 is associated with the proximal GGAA site 72 base pairs upstream of the Pax5 TSS, but not GGAA sites further from the TSS. In addition, the transcriptional repressors SIN3A and HDAC3 were detected on the same regions of the Pax5 locus. We detected association of ETV6, SIN3A, and HDAC3 with the proximal GGAA site upon expression of WT ETV6, but not ETV6 P214L. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results provide a mechanism of interaction for ETV6 and PAX5, 2 genes often disrupted in B-cell leukemia. These findings are significant because PAX5 misregulation results in a B cell development halt, lineage infidelity, and leukemogenesis. In continuing our studies, we have generated a transgenic mouse endogenously expressing the ETV6 P214L mutation by CRISPR/Cas9 editing, and these mice appear to have a thrombocytopenic phenotype similar to that observed in patients carrying the ETV6 P214L mutation. These animals will be the focus of our continued investigation of the mechanism by which ETV6 germline mutation results in a predisposition to leukemia. Our ultimate goal is a comprehensive understanding of how this process may be targeted more efficiently in patients with both heritable and sporadic forms of leukemia involving ETV6.

OBJECTIVES/SPECIFIC AIMS: The purposes of this study are to examine the relationships among sleep characteristics (duration, efficiency), stress biomarkers, and child behavior problems among toddlers living in socioeconomically disadvantaged homes and how these characteristics change over time from age of 12 months to 24 months. Aim 1: examine changes in subjective and objective sleep characteristics from 12 to 24 months of age. Aim 2: examine changes in stress biomarkers from 12 to 24 months of age. Aim 3: examine the cross sectional and longitudinal relationships between sleep characteristics and stress response. Aim 4: examine the cross sectional and longitudinal relationships between sleep characteristics and toddlers’ child behavior problems. METHODS/STUDY POPULATION: In this cross-sectional study we are recruiting parents with healthy toddlers from early head start programs and a community clinic to prospectively examine the relationships among sleep characteristics, stress biomarkers, and children’s health. Data on sleep characteristics will include subjective and objective measures of sleep duration and efficiency and parental interactive bedtime behaviors to assist their toddlers’ sleep initiation. Multisystemic biomarkers of stress including cortisol, CRP, IL-6, and BMI, will be measured individually. The associations between sleep characteristics and the biomarkers, considered as a latent variable of the stress response, will be explored. Health measures will include secretory IgA and parent-reported behavioral problems. Generalized linear models will be used in the data analysis. RESULTS/ANTICIPATED RESULTS: To date we have obtained objective (9 days/nights of actigraphy) measures of 33 toddlers’ sleep and subjective measures of parenting interactive behaviors. Using the Parental Interactive Bedtime Behavior (PIBB) Survey and subscales [active physical comforting, encourage autonomy, settle by movement, passive physical comforting (PPC), social comforting], we are currently reporting on the associations between PIBB and toddler’s sleep characteristics. The sample included 33 toddlers (mean age=1.33 years, SD=0.54). The toddlers’ sleep duration averaged 8.22 hours (SD=0.86). There were statistically significant moderate associations between sleep duration and parents’ PPC (r=−0.41, p=0.02). Intra-individual variability in the amount of wake after sleep onset was also significantly associated with total PIBB and PPC (r=0.37, p=0.05; r=0.52, p=0.002, respectively). Intra-individual variability in the amount of sleep fragmentation within toddlers was significantly associated with total PIBB (r=0.36, p=0.05). DISCUSSION/SIGNIFICANCE OF IMPACT: Although active physical comforting (eg, rocking to sleep, patting or rubbing child’s back) is most commonly associated with sleep patterns in infancy and toddlerhood among samples of higher socio-economic status, findings from this study suggest a stronger association between PPC (eg, presence of the parent in the room to fall asleep) and less sleep duration and more individual variability in night wakings. The biomarker data are currently being analyzed and results will be presented within the year. Taken together, these preliminary results and pending results will inform future intervention development that may address the role of parenting behavior in promoting health sleep early in life.

OBJECTIVES/SPECIFIC AIMS: Determine whether GABA-A receptor binding is abnormal and linked to dystonia symptoms in cervical dystonia (CD). METHODS/STUDY POPULATION: There is increasing evidence that a key pathophysiological mechanism in adult-onset focal dystonia is a reduction in inhibitory control over the sensorimotor network. Results from a recent 11C-flumazenil PET imaging study suggest that abnormal inhibitory signaling in genetic and sporadic forms of dystonia may be due to reduced GABA-A binding. It remains unknown whether CD, the most common form of adult-onset focal dystonia, is associated with abnormal GABA-A binding. The goal of this research is to determine if GABA-A receptor binding is abnormal and linked to dystonia symptoms in CD. RESULTS/ANTICIPATED RESULTS: We investigated whole brain GABA-A binding in 15 CD patients (11F; 64±8 y) and 15 healthy controls (10F; 64±9 y) using 60-minute dynamic 11C-flumazenil PET scans. GABA-A receptor binding potential (BP) was estimated using a simplified reference tissue model. A 2-sample t-test was used to identify voxel-wise GABA-A BP differences between groups, and a regression analysis used to test for correlations between GABA-A BP and disease severity as measured with the Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS). A conventional region of interest analysis was also conducted to quantify BP changes within the sensorimotor network using the automated anatomical labeling atlas. DISCUSSION/SIGNIFICANCE OF IMPACT: CD patients have reduced GABA-A receptor binding compared with healthy controls, with the greatest reduction seen within the sensorimotor region of the thalamus. Furthermore, reductions in GABA-A binding in brain regions associated with coupling sensory and motor information predict motor severity. These findings support that reduced GABAergic signaling within sensorimotor integration regions is a key mechanism underlying dystonic symptoms in CD and could help inform the development of better, more targeted treatment options.

OBJECTIVES/SPECIFIC AIMS: Mesh properties, such as stiffness, porosity, and weight have been shown to correlate with the degree of mesh integration with vaginal tissue. Previous research in rhesus macaques implanted with polypropylene mesh differing in stiffness, porosity, and weight showed differences in vaginal deterioration following mesh implantation. These differences were correlated with a foreign body response, consisting primarily of activated, proinflammatory M1 macrophages. Previous studies have determined that the early macrophage polarization profile following biomaterial implantation is a strong indicator of overall tissue integration downstream. However, these early responses have not been previously observed in the appropriate surgical models. Prior work from our laboratory in developing a cytokine delivery system has shown that shifting the macrophage response at the host-implant interface from a pro-inflammatory M1 phenotype to an anti-inflammatory M2 phenotype in the first 14 days postimplantation resulted in enhanced integration of the mesh with the surrounding tissues. The present study develops an in vivo model clinically relevant surgical model to investigate the modulation of the host response to mesh. Utilizing a moderately-sized animal, we can feasibly implant mesh using the “gold standard” abdominal sacrocolpopexy procedure and evaluate the changes in the host immunologic response at early (14 d) and tissue remodeling outcomes at late stages (90 and 180 d) of implantation. METHODS/STUDY POPULATION: Commercially available heavyweight and lightweight mesh was used to investigate the modulation of the immune response. A custom MTI SILAR Automated Dip Coating machine is used to uniformly coat the mesh in a reproducible manner. An adapted radio frequency glow discharge method is used to create a stable negative charge on the surface of the mesh, followed by the sequential deposition of polycationic and polyanionic polymers to provide a stable, conformal, nanoscale coating. Chitosan served as the polycation, chosen because of its known antimicrobial and biocompatibility properties. Dermatan sulfate served as the polyanion, chosen for its important role in regulating extracellular matrix components and enhancing the activity of cytokines. Interleukin-4 (IL-4) is incorporated into the coating to be released in a controlled manner upon implantation. In vitro controlled release profiles were assessed to demonstrate efficient and local release of IL-4. Utilizing a New Zealand white rabbit surgical model, we implant mesh using the “gold standard” abdominal sacrocolpopexy procedure and evaluate the changes in the host immunologic response at early (14 d) and tissue remodeling outcomes at late stages (90 and 180 d) of implantation. The mesh-tissue complex was removed from each rabbit and processed for histological staining as well as immunolabeling of immune cells, such as macrophages. Determination of matrix metalloproteinases and fibrotic capsule formation also helps characterize the overall inflammatory response associated with each implant. RESULTS/ANTICIPATED RESULTS: We have developed a clinically relevant rabbit surgical model to implant different conditions of surgical mesh into 2 different sites, including the vagina and the abdomen. The results of this study show that implants into vaginal tissues elicited an increased host inflammatory response at 14 days as compared with those in the abdominal wall. However, at chronic time points the inflammatory response in the vagina was reduced as compared to that in the abdominal cavity. The present study also demonstrates the scale-up of a previous methodology for nano-scale coating. We present a nanometer thickness, tunable, and uniform coating capable of releasing bioactive IL-4. In vitro assays confirm the bioactivity and the controlled local release allowing for shifts in the immune response to promote implant integration. Improved remodeling has been observed to correlate with a shift in the early host response from an M1 to an M2 phenotype, however, there is limited information on the exact mechanism. Our strategy to achieve enhanced tissue remodeling demonstrate outcomes such as minimal changes to the structural properties of the mesh and a controlled release profile to sufficiently polarize macrophages around the mesh to a pro-remodeling state. DISCUSSION/SIGNIFICANCE OF IMPACT: Pelvic organ prolapse is a condition where the pelvic floor muscles weaken over time resulting in the downward shift of the pelvic organs into the vaginal canal. Moreover, factors such as obesity, age, and vaginal birth increase the susceptibility of being diagnosed with pelvic organ prolapse. Direct costs of reconstructive procedures exceed $1 billion each year in the United States. Synthetic mesh has been used to repair abdominal hernias for over half a century. Biomedical companies, through 510k and the 1976 Medical Device Amendments Act, were able to resell their hernia repair mesh as a treatment for pelvic organ prolapse. However, women who have had vaginal mesh implants have reported an increasing number of complications including chronic pain and mesh erosion/exposure at rates as high as 10%–20%. In fact, in 2008 and 2011, the US Food and Drug Administration issued warnings to doctors and patients about the mesh. In January 2016, the FDA officially had to reclassify surgical mesh for transvaginal repair of pelvic organ prolapse from a class II, moderate risk device, to a class III, high-risk device. Presently, data for the use of synthetic mesh has largely derived from abdominal hernia repair, instead of vaginal repair of prolapse. In the rodent model, the vagina is too small to implant mesh in an analogous manner to human implantation. Instead, implantations are done in the abdomen, a different tissue composition and host response profile than the vagina. Primate models of pelvic organ prolapse have been utilized, but are associated with high costs and investigation of acute immune responses are not considered ethical due to the short time of survival. Thus, our presented work will not only show the development of an improved material for implantation, but also the development of an in vivo model clinically relevant to understanding the early host response to mesh.

OBJECTIVES/SPECIFIC AIMS: To establish an in vitro quantitative method for the evaluation of polymeric film disintegration that can be applied to predict in vivo behavior. METHODS/STUDY POPULATION: Two clinically advanced vaginal microbicide film products containing tenofovir and dapivirine were used as model films throughout this work. Films were made using the solvent cast manufacturing method in which polymers, excipients, plasticizer, and APIs were either dissolved or dispersed in water, mixed, and cast on a heated substrate. The novel, quantitative method was developed using a TA.XT Plus Texture Analyzer® (Texture Technologies) in combination with a TA-108S5 fixture and the TA-8A: 1/8″ diameter rounded end ball probe. Exponent® was used as the data analysis software. In this method, the film was placed and secured in the fixture, the probe applied a constant force to the film product, and a biologically relevant amount of fluid was applied to the film. The probe was able to penetrate the film upon disintegration resulting in an applied force of zero at that point. A curve of force Versus time was plotted, and disintegration time was defined as the time between fluid addition until the probe force reached zero. Test parameters were optimized in order to reduce error. Visual observation of film disintegration was conducted in the in vivo macaque model using films that included a water-soluble blue dye for film visualization. Colpophotography was also used to confirm film disintegration. In vitro results were compared with in vivo findings. RESULTS/ANTICIPATED RESULTS: The Texture Analyzer disintegration method developed provided quantitative disintegration times and did not rely on user defined endpoints which is common in many visual disintegration tests. The disintegration method was able to distinguish differences between the 2 clinical film products and produced reproducible disintegration times for the tenofovir and dapivirine films. The tenofovir film had a shorter disintegration time (41.28±2.85 s) compared with that of the dapivirine film (88.3 6±9.82 s). This method was also able to distinguish changes made to these 2 clinical film products in terms of volume and formulation alterations. In vitro and in vivo disintegration times differed by orders of magnitude, with in vitro time being measured in seconds and in vivo time being measured in days, for a variety of factors, mainly the application of constant force to the film product. Regardless of these differences, the rank order of film disintegration remained constant for in vitro and in vivo disintegration and an In Vitro In Vivo Correlation (IVIVC) trend could be seen. DISCUSSION/SIGNIFICANCE OF IMPACT: Standardization of preclinical in vitro assessments which minimize user bias are crucial to the field of pharmaceutical film development. As this field continues to develop and more products advance for pharmaceutical application, this method has the potential to become a standard assessment of film functionality. This study represents a first step in the process of developing an IVIVC. More films will need to be tested using both in vitro and visual methods in order to establish and accurate factor to predict in vivo behavior.

OBJECTIVES/SPECIFIC AIMS: To determine the association between clinical characteristics and platelet reactivity in Hispanic patients on clopidogrel therapy. METHODS/STUDY POPULATION: A cross-sectional pilot study was performed in 58 Puerto Rican patients diagnosed with any type of vascular disease and actively receiving a maintenance dose of clopidogrel for at least 7 days. The study population was divided into 2 groups: Group I with non-high on-treatment platelet reactivity (TPR); Group II with high TPR. To determine the platelet function, P2Y12 reaction units (PRU) were obtained by VerifyNow® P2Y12 assay (Accumetrics, USA). RESULTS/ANTICIPATED RESULTS: We studied a heterogeneous cohort of patients with coronary artery disease (57%), peripheral artery disease (30%), carotid artery stenosis (7%), cerebral artery aneurysm (3%), and stroke (3%) on clopidogrel therapy for secondary prevention of thromboembolic events. The mean TPR was 205±49 PRU (range: 61–304), with a prevalence of 28% patients with high TPR (PRU≥230). No significant clinical differences were found between the non-high TPR and high-TPR groups (p>0.05). However, multivariable logistic regression analysis showed that both diabetes mellitus (OR=7.5; CI: 1.01–51.9) and proton-pump inhibitors (OR=13.6; CI: 1.3–142.0) were independently correlated with high TPR (p<0.05) after adjusting for other clinical variables. DISCUSSION/SIGNIFICANCE OF IMPACT: These results provide new insight into the importance of clinical characteristics on platelet reactivity in this Caribbean population. Further studies are warranted to determine whether important clopidogrel pharmacogenes are related with platelet function in Hispanics, as well as the role of TPR in guiding antiplatelet therapy and predicting future adverse cardiovascular events in this population.

OBJECTIVES/SPECIFIC AIMS: To establish a state-wide research registry of diverse participants. METHODS/STUDY POPULATION: We garnered broad institutional and community support by involving TRI’s Community Engagement team, its Community Advisory Board (CAB), and 3 UAMS patient CABs in selecting Web site content, images, and colors. Using this feedback, the TRI Recruitment Unit (RU), in conjunction with UAMS Communications and the Center for Health Literacy, developed the materials and crafted comprehensive communication and recruitment strategies. The UAMS Center for Pacific Islander Health, Hispanic faculty, and CAB members translated materials. UAMS IT programmed the user-friendly site to allow registration from smartphones and i-Pads and linked to UAMS patient electronic health messages. RESULTS/ANTICIPATED RESULTS: The RU committee implemented successful innovative strategies, including recruiting at the Arkansas State Fair and ballgames, attended by people of all races, ages, and socio-economic levels. Using i-Pads at the sites, recruitment took <5 minutes/registrant. Within 8 months, >2400 participants from across Arkansas had joined the registry: 14% African-Americans, 8% Pacific Islanders, 5% Hispanic, and 3% Native American. DISCUSSION/SIGNIFICANCE OF IMPACT: Involving CAB multidisciplinary input to design and implement recruitment materials was highly successful. Despite challenges of recruiting under-represented groups, the registry includes 30% minorities. By tracking registrants’ demographics with Lime Survey software, the RU will prioritize future recruitment events to maximize diversity of registrants.

OBJECTIVES/SPECIFIC AIMS: (1) Determine person (sex, age, education level), environment (marital status, living alone, insurance), and health and illness (BMI, type of AF, comorbidities) characteristics that are associated with outcomes (QOL, symptom severity, and emotional and functional status). (2) Determine the association of symptom management strategies (ablation, cardioversion, and rate and rhythm control medications) and outcomes (QOL, symptom severity, and emotional and functional status). (3) Test person (sex, age, and education level) and environment (marital status, living alone, insurance) characteristics as moderators of the effect of symptom management strategies (ablation, cardioversion, and rate and rhythm control medications) on outcomes (QOL, symptom severity, and emotional and functional status). METHODS/STUDY POPULATION: AF patients (≥18 years of age) already enrolled in the PaTH study will be included. To date, 1026 total participants have been enrolled. Based on the enrolled participants, 92% (945) of our study population are Caucasian and 36% (362) are female. The age range of the enrolled participants is: 2% (16) 18−39, 4% (42) 40−49, 11% (108) 50−59, 33% (343) 60−69, 34% (353) 70−79, and 16% (162) 80+. Participants are recruited through in-person, email, phone, patient portal messaging and post mail techniques to ensure a representative sample. The PaTH study integrates electronic health record and insurance claims data with patient-reported outcome measures collected through online surveys. RESULTS/ANTICIPATED RESULTS: We hypothesize that sex, older age, low education level, living alone, absence of partner, absence of insurance coverage, high BMI, and a high number of comorbidities will be associated with lower QOL, high symptom severity, and low emotional and functional status. We further hypothesize that symptom management strategies will be associated with higher QOL, low symptom severity, and high emotional and functional status, and that these associations will be moderated by person and environment characteristics. DISCUSSION/SIGNIFICANCE OF IMPACT: The proposed research is an important first step in determining potential causes of person and environment differences in symptom severity. It will lead to tailored symptom management interventions for individuals most at risk for experiencing high symptom severity.

OBJECTIVES/SPECIFIC AIMS: Aim 1—estimate prevalence and associated characteristics of nonfatal, non-self-inflicted strangulation among women ages 18 and older who presented to a US emergency department between 2006 and 2013. Aim 2—explore care-seeking behaviors, the context of the care seeking, treatment expectations and perceived diagnosis in a sample of women ages 18 and older who present to a US emergency department and report being strangled by an intimate partner. Aim 3—merge and synthesize findings from both the quantitative and qualitative strands to provide a more complete understanding of post-strangulation emergency care of women. METHODS/STUDY POPULATION: This mixed-methods study will use a convergent parallel design, with a single phase of concurrent and independent data collection. Analysis of quantitative and qualitative data will be performed separately then compared, with main findings integrated during the interpretation phase and presented in a merged data analysis display. IRB review and approval will be obtained before initiating this study. Aim 1 will include a cross-sectional analysis of 2006–2013 Nationwide Emergency Department Sample (NEDS) data, from the Agency for Healthcare Research and Quality’s Healthcare Cost and Utilization Project (HCUP). NEDS is the US’s largest all-payer emergency department (ED) database, providing national estimates of hospital-based ED visits from ~120 to 135 million ED visits/year (weighted). For this study, we will examine data from patients meeting inclusion criteria with an International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM; Medicode, 1996) code of strangulation. For this strand, females aged 18 years or older who presented to a US emergency department between 2006 and 2013 will be included. The outcome variable will be non-fatal, non-self-inflicted strangulation, defined using at least one of the ICD-9-CM codes for strangulation. These codes are: 994.7 (“asphyxiation and strangulation”), E963 (“assault by hanging and strangulation”), E983.8 (“strangulation or suffocation by other specified means undetermined whether accidentally or purposely inflicted”), and E983.9 (“strangulation or suffocation by unspecified means undetermined whether accidentally or purposely inflicted”). Patients with a concurrent ICD-9-CM code for suicide attempt (E953, “Suicide and self-inflicted injury by hanging, strangulation and suffocation”) will be excluded, to minimize self-inflicted assault events. Aim 2 will employ a narrative descriptive approach, with semistructured individual interviews to gather more information about women’s experiences when engaging the health care system after being strangled. Medical records related to the strangulation event will also be reviewed for diagnostic codes and other nursing and/or medical notes that may relate to diagnoses, treatment and referrals. For this strand, women aged 18 years or older who present for care to an urban, academic ED will be recruited, purposely sampling those reporting strangulation as a reason for their visit. We anticipate interviewing ~20–30 women to achieve saturation of information. RESULTS/ANTICIPATED RESULTS: Data from the NEDS from 2006 to 2013 will be analyzed for prevalence and associated characteristics of women seeking care after being strangled. Individual interviews and medical record reviews of a small sample of adult women will be conducted to explore women’s in-depth experiences within the health care system. Results from both the quantitative and qualitative analyses will then be collectively compared and interpreted to better synthesize the evidence from this work. Convergent and divergent findings will be presented in a merged data analysis display (Creswell and Plano Clark, 2011). Qualitative data will be used to fill the knowledge gap remaining from the quantitative analysis, and to explain and contextualize some of the findings. Such integration will help expand the current limited evidence on care of strangled women, and will identify additional research questions that will guide future research in this area. DISCUSSION/SIGNIFICANCE OF IMPACT: To our knowledge, this study will be the first to explore this issue using a nationally representative sample of adult women who sought emergency medical care for strangulation analyzed in conjunction with a detailed qualitative analysis of strangled women’s experiences with the health care system. The resulting knowledge will be critical to informing clinical assessment, intervention and prevention efforts for this vulnerable population, as well as public policy and future research regarding this specific violence tactic.

OBJECTIVES/SPECIFIC AIMS: Improving public health requires effective community-engaged approaches. The Better Together Lebanon County initiative plans to create opportunities for improved health and quality of life by aligning strategies of local organizations, previously working independently. METHODS/STUDY POPULATION: Better Together began with a 1-day summit, convening stakeholders with the goal of coordinating efforts and maximizing resources in the Lebanon community. Key stakeholders were identified using the socioecological model to assist with planning, goal setting, and developing outcomes for this initiative. Representation included community members, hospital systems, restaurants, school administrators, nonprofit organizations (including YMCA, American Heart Association), grocery stores and policy makers such as the mayor, health departments, and state representatives. RESULTS/ANTICIPATED RESULTS: The Better Together summit brought together almost 200 individuals representing 82 local organizations to share ideas and evoke collaboration around decreasing health disparities. Attendees learned about programs within and outside of their communities and volunteered for task forces to propel the community forward. Currently, we have members committed to further this work through Action Teams within the sectors of Physical Activity, Healthy Food Access and Family and Community Engagement. DISCUSSION/SIGNIFICANCE OF IMPACT: Convening individuals from many layers of the community helps to ensure discussions are representative of the overall community voice. It is vital to facilitate effective collaboration that includes networking, identifying assets and areas of improvement, brainstorming solutions and integrating research and best practices to improve the health of a community.

OBJECTIVES/SPECIFIC AIMS: Chromosome 22q11.2 deletion syndrome (22q) has a prevalence almost as common as Down syndrome. 22q is well known for medical complications, including congenital heart disease and immune dysfunction. However, children with 22q also have borderline cognitive abilities, are at high risk for ADHD and anxiety, and have poor independent living skills (adaptive function). Parenting is one modifiable factor that has been found in typically developing populations to promote independent functioning and protect against the development of anxiety disorders. This study investigates the associations between parenting, anxiety, and adaptive functioning in 22q. METHODS/STUDY POPULATION: Parent-child (ages 4–11) dyads participated in an ongoing study involving observed parenting during challenging tasks plus questionnaires of parenting, child anxiety, and child functioning. In total, 52 dyads [22q=25; typical development (TD)=27] have enrolled to date. Parents completed questionnaires, including the Parenting Styles and Dimensions Questionnaire (PSDQ), Spence Children’s Anxiety Scale, and Adaptive Behavior Assessment System for Children (ABAS-II). PSDQ dimensions of interest included Parental Psychological Control (PPC: the management of child behavior through the manipulation of emotions, expectations, and independence), Authoritative, Authoritarian, and Permissive, and the subscales of these broad dimensions. Scores were compared using t-tests and multiple regression models were used to investigate the relationships between 1-parenting and anxiety and 2-parenting and adaptive function. RESULTS/ANTICIPATED RESULTS: Mean age was 7.8+2.1 years. Full Scale IQ (TD: 112.3 vs. 22q: 82; p<0.001) and ABAS-II Global Adaptive Composite (TD: 102.7 vs. 22q: 69.2; p<0.001) were significantly higher in the TD group. Parents in the 22q group reported higher levels of PSDQ PPC (22q: 2.3 vs. TD: 2.1; p=0.06), specifically overprotection (22q: 3.7 vs. TD: 3.3; p=0.04), and lower Authoritative parenting (22q: 4.1 vs. TD: 4.4; p=0.03), across the subscales. There were no differences in Authoritarian or Permissive parenting. Children with 22q had higher Spence Total Anxiety scores (22q: 62.5 vs. TD: 47.4; p<0.001). Self-reported PPC and group (R2=0.3, F3,48=8.1, p<0.001) predicted child anxiety with a main effect of PPC (β=16, p=0.02). Group tended to moderate the association between PPC and anxiety (β=−17.5, p<0.10), with PPC predicting anxiety for the 22q group (r=0.35, p<0.09), but not the TD group (r=−0.08, ns). At this time, a relationship between PPC and child ABAS-II GAC in 22q (r=−0.14; p=0.5) is not identified. DISCUSSION/SIGNIFICANCE OF IMPACT: Children with 22q are at high risk for anxiety and poor adaptive outcomes. These results suggest that parents of children with 22q use higher levels of PPC, which is correlated with increased child anxiety. These analyses also provide support for parenting interventions to improve anxiety in children with 22q and possibly mitigate the serious mental health risk in this population.

OBJECTIVES/SPECIFIC AIMS: This study seeks to test the feasibility and effectiveness of a brief acceptance and commitment therapy (ACT) treatment for chronic pain patients in a primary care clinic METHODS/STUDY POPULATION: Primary care patients aged 18 years and older with at least 1 pain condition for 12 weeks or more in duration will be recruited. Patients will be randomized into (a) ACT intervention or (b) control group. Participants in the ACT arm will attend 1 individual visit with an integrated behavioral health provider, followed by 3 weekly ACT classes and a booster class 2 months later. Control group will receive enhanced primary care that includes patient education handouts informed by cognitive behavioral science. Data analysis will include 1-way analysis of covariance (ANCOVA), multiple regression with bootstrapping. RESULTS/ANTICIPATED RESULTS: The overall hypothesis is that brief ACT treatment reduces physical disability, improves functioning, and reduces medication misuse in chronic pain patients when delivered by an integrated behavioral health provider in primary care. In addition, it is anticipated that improvements in patient functioning will be mediated by patient change in pain acceptance and patient engagement in value-consistent behaviors. DISCUSSION/SIGNIFICANCE OF IMPACT: This pilot study will establish preliminary data about the effectiveness of addressing chronic pain in a generalizable integrated primary care setting. Data will help support a larger trial in the future. Findings have potential to transform the way chronic pain is currently managed in primary care settings, with results that could decrease disability and improve functioning among patients suffering from chronic pain.

OBJECTIVES/SPECIFIC AIMS: Thionamides are anti-thyroid drugs (ATD) that are commonly used to treat autonomous thyrotoxicosis. Although efficacious, these medications carry a risk of neutropenia or agranulocytosis in a small but finite proportion of the patients who receive them. Some risk factors for thionamide-induced neutropenia have been identified, including body mass index (BMI) and dose, but the role of race and ethnicity in the pathogenesis of this potentially life-threatening side effect is not known. We hypothesize that there will be no effect of race or ethnicity on the change in absolute neutrophil count (ANC) following initiation of thionamide therapy among adult patients with thyrotoxicosis. METHODS/STUDY POPULATION: Data from the electronic medical record at UNM HSC were obtained using a standard database query for the years 2000–2016. Inclusion criteria were the prescription of an ATD, an ANC recorded within 30 days of initiating ATD therapy (pre-ATD), and an ANC recorded between 75 and 365 days after starting an ANC (post-ATD). Patients taking other agents known to cause neutropenia and agranulocytosis, such as clozapine, allopurinol, or chemotherapy, were excluded. Patients were assigned to racial and ethnic groups as follows: Hispanic, non-Hispanic Caucasian (NHC), native American, Black, and Asian. The post-ATD ANC was defined as the nadir ANC observed after the ATD was started. “Delta ANC” was defined as [(post-ATD ANC)−(pre-ATD ANC)]. ANOVA analysis with Bonferroni-adjusted post-hoc testing was performed to examine differences in the mean changes of ANC across ethnic groups. RESULTS/ANTICIPATED RESULTS: In total, 123 adult patients met inclusion and exclusion criteria and were included in the analysis. No significant difference was found between any of the racial groups with regard to age, sex, BMI, pre-ATD ANC, or the pre-ATD to post-ATD ANC interval. The native American group showed a significantly greater post-ATD ANC (not shown) and Delta-ANC as compared with the other groups. Delta ANC Hispanic=−1.4±3.3, Caucasian=−0.6±3.3, Black=−0.9±4.1, Asian=−3.8±4.8, native American=3.6±5.1 (all units per mm3; p<0.001). DISCUSSION/SIGNIFICANCE OF IMPACT: In this cohort of New Mexicans with thyrotoxicosis, native American race was protective against thionamide-induced neutropenia.

OBJECTIVES/SPECIFIC AIMS: To investigate whether medical complexity (indicated by multiple providers or healthcare visits) is associated with lower levels of confidence in medication use and lower medication adherence METHODS/STUDY POPULATION: Data on socio-demographics, health encounters, health status, and health attitudes and behaviors from a nationally representative sample of 1575 older Singaporean adults were utilized. The association of medical complexity factors with self-reported medication confidence and adherence was analyzed using logistic regression analysis controlling for age, gender, ethnicity, education, and number of health conditions. RESULTS/ANTICIPATED RESULTS: The survey had a 60% response rate. The mean age of respondents was 72, and 42% were male. We found no significant association between number of visits and either confidence about usage (OR=1.07, 95% CI 0.95–1.20) or medication adherence (OR=1.01, 95% CI 0.90–1.13). We similarly found no significant association between number of providers and either confidence about usage (OR=1.03, 95% CI 0.90–1.18) or medication adherence (OR=1.05, 95% CI 0.93–1.20). Lower confidence about medication use was less likely among males (OR=0.60, 95% CI 0.44–0.80), those with more education (OR=0.29, 95% CI 0.20–0.42) or more comorbidities (OR=0.89, 95% CI 0.82–0.96) and more likely with increasing age (OR=1.06, 95% CI 1.04–1.08). Nonadherence was more likely among Indians (OR=1.62, 95% CI 1.06–2.48) and those with more comorbidities (OR=1.10, 95% CI 1.02–1.18). DISCUSSION/SIGNIFICANCE OF IMPACT: Having more healthcare visits or providers were not independent correlates of lower medication confidence or adherence. Seniors with less education may benefit from interventions to improve confidence about medication use. Participants with more comorbidities expressed greater confidence but admitted to lower adherence. The role of other potential contributors to nonadherence in complex patients (eg, cost and access, patient preference, competing demands) should be evaluated next.

OBJECTIVES/SPECIFIC AIMS: The goal of this pilot study is to provide a reliable anatomical algorithm for the measurement of adipose tissue within the pelvic cavity as a predictor of prostate cancer aggressiveness and recurrence after radical prostatectomy. METHODS/STUDY POPULATION: We will conduct a retrospective analysis of men treated with radical prostatectomy between 2012 and 2016 at the VA Caribbean Health Care System. Clinical variables, pathology reports, and computed tomography will be reviewed. Pelvic and periprostatic fat (PF) will be measured to determine association between PF and cancer aggressiveness and recurrence. RESULTS/ANTICIPATED RESULTS: We expect a positive association between PF and cancer aggressiveness and recurrence among patients who underwent radical prostatectomy. DISCUSSION/SIGNIFICANCE OF IMPACT: Measurement of subcutaneous and PF within the pelvic cavity can provide a reliable anatomical measure which can be used as a proxy measure to identify those with higher risk of recurrence and develop better prevention and treatment strategies, especially in Hispanic men.

OBJECTIVES/SPECIFIC AIMS: Inflammatory bowel disease (IBD) patients are at an increased risk of Clostridium difficile infection (CDI) but the impact of CDI on disease severity is unclear. The aim of this study was to determine the effect of CDI on long-term disease outcome in a cohort of IBD patients. METHODS/STUDY POPULATION: We analyzed patients enrolled in a prospective IBD natural history registry. Patients who tested positive at least once formed the CDI positive group. We generated a 2:1 propensity matched control cohort based on risk factors of CDI in the year before infection. Healthcare utilization data (emergency department use, subsequent hospitalizations, telephone encounters), medications, labs, disease activity, and quality of life metrics were temporally organized. RESULTS/ANTICIPATED RESULTS: A total of 198 patients (66 CDI, 132 matched controls) were included [56.6% female; 60.1% Crohn’s disease (CD), 39.9% ulcerative colitis (UC)]. Groups were not significantly different in the year before infection in all metrics but in the year of infection, having CDI was significantly associated with more steroid and antibiotic exposure, elevated C-reactive protein or erythrocyte sedimentation rate, and low vitamin D (all p<0.01). Infection was associated with increased disease activity metrics (UC: p=0.036, CD: p=0.003), worse disease-related quality of life (p=0.003), and increased healthcare utilization (p<0.001). In the next year after infection those with prior CDI continued to have increased exposure to vancomycin or fidaxomicin (p<0.001) and all other antibiotics (p=0.01). They also continued to have more clinic visits (p=0.006), telephone encounters (p=0.001), and worse disease-related quality of life (p=0.03), but disease activity and biomarkers of severity were not significantly different between groups. DISCUSSION/SIGNIFICANCE OF IMPACT: CDI infection in IBD is significantly associated with various surrogate markers of disease severity, increased healthcare utilization and poor quality of life during the year of infection. CDI patients continue to experience poor quality of life after infection with increased clinic visits and antibiotic exposure while disease activity is no longer significantly increased. These findings suggest that CDI infection may have a lasting effect on healthcare utilization beyond the acute treatment period.

OBJECTIVES/SPECIFIC AIMS: Although the clinical utility of whole genome sequencing (WGS) is increasing, a gap exists between what WGS can deliver in quantity of genomic information and what results can be interpreted that patients and community members would find meaningful. Given the potential for incidental findings and variants of uncertain significance, an emphasis should be placed on understanding patient preferences towards receiving WGS results. To identify the current knowledge base on WGS preferences, we performed a scoping review. METHODS/STUDY POPULATION: A search on PubMed using terms “WES,” “WGS,” “genome sequencing,” ”attitudes,” and “preferences” identified survey research between 2012 and 2016. Summaries of population, sample, variables, and results were tabulated. RESULTS/ANTICIPATED RESULTS: Of 13 studies identified, 6 surveyed community members, 6 included medical professionals, and 2 surveyed cohorts with a specific medical condition. Only 1 study used a nationally representative sample and no study focused on a medically underserved population. Patients were most interested in receiving medically actionable results, yet preferred to have access to all available data if desired. Genetics professionals are more conservative with the return of incidental and uncertain findings. DISCUSSION/SIGNIFICANCE OF IMPACT: Existing surveys have limited representation of the US public. Future studies focused on medically underserved populations would provide a deeper understanding of attitudes and preferences toward WGS.

OBJECTIVES/SPECIFIC AIMS: To build a multisite deidentified database of female adolescents, aged 12–21 years (January 2011–December 2012), and their subsequent offspring through 24 months of age from electronic health records (EHRs) provided by participating Community Health. METHODS/STUDY POPULATION: We created a community-academic partnership that included New York City Community Health Centers (n=4) and Hospitals (n=4), The Rockefeller University, The Sackler Institute for Nutrition Science and Clinical Directors Network (CDN). We used the Community-Engaged Research Navigation model to establish a multisite deidentified database extracted from EHRs of female adolescents aged 12–21 years (January 2011–December 2012) and their offspring through 24 months of age. These patients received their primary care between 2011 and 2015. Clinical data were used to explore possible associations among specific measures. We focused on the preconception, prenatal, postnatal periods, including pediatric visits up to 24 months of age. RESULTS/ANTICIPATED RESULTS: The preliminary analysis included all female adolescents (n=49,292) and a subset of pregnant adolescents with offspring data available (n=2917). Patients were mostly from the Bronx; 43% of all adolescent females were overweight (22%) or obese (21%) and showed higher systolic and diastolic blood pressure, blood glucose levels, hemoglobin A1c, total cholesterol, and triglycerides levels compared with normal-weight adolescent females (p<0.05). There was a statistically significant association between the BMI status of mothers and infants’ birth weight, with underweight/normal-weight mothers having more low birth weight (LBW) babies and overweight/obese mothers having more large babies. The odds of having a LBW baby was 0.61 (95% CI: 0.41, 0.89) lower in obese compared with normal-weight adolescent mothers. The risk of having a preterm birth before 37 weeks was found to be neutral in obese compared to normal-weight adolescent mothers (OR=0.81, 95% CI: 0.53, 1.25). Preliminary associations are similar to those reported in the published literature. DISCUSSION/SIGNIFICANCE OF IMPACT: This EHR database uses available measures from routine clinical care as a “rapid assay” to explore potential associations, and may be more useful to detect the presence and direction of associations than the magnitude of effects. This partnership has engaged community clinicians, laboratory and clinical investigators, and funders in study design and analysis, as demonstrated by the collaborative development and testing of hypotheses relevant to service delivery.

OBJECTIVES/SPECIFIC AIMS: (1) Provide an innovative tool used to accelerate and evaluate T3-T4 research; (2) describe the collective capacity building tool (CCBT) methodology—both programmatic and evaluative applications; and (3) share insights about the process and outcomes of community-engaged research. METHODS/STUDY POPULATION: Academic and community-based partners complete the assessment together at the beginning and conclusion of their Community Engagement pilot projects. Further, they are encouraged to use the tool and the associated insights/priorities that emerge as the basis for data-driven coaching with Community Research Liaisons throughout the 12-month grant cycle. RESULTS/ANTICIPATED RESULTS: Pre/post results with 4 cohorts of pilot grantees consistently demonstrated the most positive change in relation to 1 item: overcoming previously identified barriers to community engagement (eg, language, mistrust, scheduling conflicts). Other key findings: (1) networks of reciprocal ties expand, providing structures to support dissemination of information and interventions. (2) Partners leverage expanded networks to pursue follow-on funding and extend the scope/reach of their efforts geographically and/or with new populations. (3) Projects enhance trust in the research process by developing group processes that facilitate the respectful sharing of diverse (often alternative) viewpoints and through culturally-responsive project implementation. DISCUSSION/SIGNIFICANCE OF IMPACT: The CCBT can be used at multiple points in time to help project partners achieve the deliberate integration of CBPR principles in practice and advance community-engaged translational research efforts for sustainability and scalability. The CCBT is sensitive enough to document the iterative nature of partnership development and CBPR. An example: a great deal of variability was found in how formally partners defined roles. Further, partner roles often changed as projects evolved. Still, results indicated a general trend toward achieving greater clarity in partner roles over time. Further, the tool captured set-backs due to partner turn-over and partnerships regaining momentum after new staff came on board. Results have strong face validity: more mature partnerships reported stronger community connections and previous successes to build upon. Perhaps most importantly: the tool and associated process was well-received by academic and community-based partners alike.