Translational Science 2013 Abstracts
Version of Record online: 18 APR 2013
© 2013 Wiley Periodicals, Inc.
Clinical and Translational Science
Volume 6, Issue 2, pages 103–164, April 2013
How to Cite
(2013), Translational Science 2013 Abstracts. Clinical and Translational Science, 6: 103–164. doi: 10.1111/cts.12047
- Issue online: 18 APR 2013
- Version of Record online: 18 APR 2013
- Top of page
- SCHOLAR/TRAINEE ABSTRACTS
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
- Research Professionals Abstracts Best Practices
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
AUTHOR'S CLINICAL SPECIALTY: A COMPETING INTEREST IN ECONOMIC ANALYSIS OF SURGICAL INTERVENTIONS; REALITY OR FICTION?
Alawami EM1, Ruthazer R3,4, Cohen JT2,3, Kent DM1,4 1Tufts Clinical and Translational Science Institute - Tufts Medical Center, Boston, MA, USA; 2Center for the Evaluation of Value and Risk in Health, Boston, MA, USA; 3Institute for Clinical Research and Health Policy Studies - Tufts Medical Center, Boston, MA, USA; 4Tufts University School of Medicine, Boston, MA, USA
OBJECTIVES/SPECIFIC AIMS: To determine if author's clinical specialty is associated with a favorable cost effectiveness ratio (CER) when examining the reported primary CER for surgical interventions in the CEA registry. METHODS/STUDY POPULATION: We used the Tufts Medical Center Cost Effectiveness Analysis Registry (www.cearegistry.org); a comprehensive database of cost-utility analyses that have been performed for a wide variety of diseases conditions and treatment options. All articles and ratios labeled as surgical in the CEA registry published between (1976–2011) were included. Logistic regression models were used to examine the multivariate relationship between the primary outcome (favorable ratios) and the primary author's clinical specialty; while controlling for the source of funding and comparator category. RESULTS/ANTICIPATED RESULTS: Preliminary results show cost analyses comparing surgical interventions with alternative treatment options tend produce more favorable cost effectiveness ratios -compared to the weighted median CER in our sample- when the primary author is a surgeon. DISCUSSION/SIGNIFICANCE OF IMPACT: By examining the association between authors' clinical specialty and favorable cost effectiveness ratios, we attempt to determine if more efforts should be directed to control for non-financial competing interests stemming from the author's clinical bias and provide a guided prospective for stakeholders to utilize this vast body of literature in different interlinking processes, including policy making, clinical practice guidelines development, and reimbursement for services.
CLINICIAN KNOWLEDGE, ATTITUDE, AND BEHAVIOR REGARDING BEST PRACTICES FOR PROVIDING HIV/AIDS CARE AND DISCLOSING POSITIVE HIV STATUS TO SEXUAL/ NEEDLE-SHARING PARTNERS IN GEORGIA
Ede V1, Yancey E1, Stringer HG1, Josiah-Willock R1, Strayhorn G1 1Morehouse School of Medicine, Atlanta, GA, USA
OBJECTIVES/SPECIFIC AIMS: (a) To assess the knowledge and (b) attitudes, of Georgia clinicians on best practices for providing HIV/AIDS care and disclosure of positive HIV status to sexual and needle-sharing partners (c) To describe the clinical practices of Georgia clinicians regarding HIV/AIDS care and disclosure of positive HIV status to sexual and needle-sharing partners. METHODS/STUDY POPULATION: Cross-sectional survey of 100 HIV care providers in Georgia who were more likely to serve minority and underserved populations. Chi-square was used to compare knowledge,behavioral and attitudinal responses. Logistic regression was used to test the relationship between provider behavior, attitude, and knowledge regarding best practices for HIV care and status disclosure to at-risk partners. RESULTS/ANTICIPATED RESULTS: 48% of HIV care providers had poor behavior scores. 56% of providers revealed less positive attitudes. There was considerably low (27%) knowledge about Georgia law on HIV status disclosure among clinicians. Only 32% of the providers had earned some HIV-related CME credits within the past 2 years. The odds of good behavior among providers with HIV-related CME credits were 8 times greate The odds of good behavior among providers who had positive attitudes toward best practice guidelines were 24 times that of those with less positive attitudes (OR; 24.4, 95%CI, 3.6–167.5, P = 0.001). DISCUSSION/SIGNIFICANCE OF IMPACT: Further research that targets at ways to improve clinicians' knowledge levels, attitudes, and behaviors regarding best clinical practices for providing HIV care and disclosing HIV status to at-risk partners of HIV-infected persons, is needed to address high HIV incidence rates.
EVALUATION OF TWO MEASUREMENTS FOR CROWN MARGIN PLACEMENT ON GINGIVAL HEALTH AND AESTHETICS
Murillo M1, Fernandez O2, Elias AR1 1University of Puerto Rico, Medical Sciences Campus, San Juan, USA; 2University of Costa Rica, San Jose, Costa Rica
OBJECTIVES/SPECIFIC AIMS: The dentogingival complex (DGC) dimension's integrity is indicative of gingival health, and a guide for subgingival crown margin placement. Defining the DGC dimension when preparing subgingival margin on anterior teeth, allows the dentist to determine the optimal position of margin placement, preventing effects on gingival health and aesthetics leading to gingival inflammation, attachment loss, and bone loss. There are two measurements to establish the DGC dimension in restorative dentistry. Gingival sulcus (GS) measurements based on free gingiva and X-ray and the DGC. There is a dilemma as to which measurement is more effective in preventing adverse events on the gingival tissue when establishing crown margin placement. In this study we propose that the DGC measurement will better establish the crown margin placement resulting in healthy gingival tissue and better aesthetics than the GS measurement. METHODS/STUDY POPULATION: We propose to evaluate and compare the DG and GS measurement around premolar, canine, and incisor teeth in the maxillary and mandibular arches as measured by transulcus probing. We will compare the gingival health and aesthetics after six months of having crown placement employing the DG or the GS measurements. RESULTS/ANTICIPATED RESULTS: Pending DISCUSSION/SIGNIFICANCE OF IMPACT: We will gain information relevant to the dentist that support the evidence that the DG measurement, is key to prevent and maintaining the gingival health and aesthetic problems. “This work was partially supported by the NIH and Health Disparities of the National Institutes of Health under Award Number R25MD007607. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.”
IMPROVING BLOOD PRESSURE CONTROL IN PEDIATRIC KIDNEY TRANSPLANT RECIPIENTS
Hooper DK1,2, Margolis P1,2 1Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA; 2University of Cincinnati, Cincinnati, OH, USA
OBJECTIVES/SPECIFIC AIMS: Despite available therapies, 50% of pediatric kidney transplant recipients (KTRs) have uncontrolled hypertension (HTN) which shortens allograft survival by 50%. Our objective was to improve blood pressure (BP) control in a large pediatric KT program with 10 providers at 5 clinic locations. METHODS/STUDY POPULATION: Using quality improvement methods we standardized BP measurement, documentation and classification during clinic visits using the following interventions: education, decision aids, performance tracking and feedback, decision support, and pre-visit planning. Our primary outcome was the percentage of patients >3 mos. post-KT with BP controlled at their most recent visit. Controlled BP was defined as BP <120/80 and <90th percentile for age, gender, height for KTRs <18 y.o. and <130/80 for KTRs ≥18 y.o.. We also measured visits where BP was recorded correctly according to 8 criteria and visits where BP was classified by providers. We used statistical process control methods to analyze data over time starting 5 mos. prior to interventions and continuing for 2 years. Data points outside 3-sigma control limits reflected significant change. RESULTS/ANTICIPATED RESULTS: BP control improved from 48% to 63% of the population over 1 year. BP was documented properly in >90% of visits within 2 months and classified in >90% of visits within 7 months. Special cause variation indicated significantly improved population BP control within 4 months of reliable BP documentation, classification, and pre-visit planning. DISCUSSION/SIGNIFICANCE OF IMPACT: Standardized measurement and recognition of HTN led to rapid, significant improvement in BP control. We expect that more comprehensive system redesign including standardized treatment and self-management support will lead to greater BP control for KTRs and improve cardiovascular and renal outcomes in these patients.
THE AUTISM MENTAL STATUS EXAM: VALIDATION IN A COMMUNITY-BASED POPULATION
Grodberg D1 1Seaver Autism Center, Mount Sinai School of Medicine, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: The diagnosis of autism spectrum disorders (ASD) in underserved and under-resourced populations has been rendered unreliable by the lack of a streamlined and standardized approach towards the observation and documentation of ASD signs and symptoms. Consequently there is a significant delay in ASD diagnosis: although parents first report concerns when their child is 18 months of age, the average age children receive ASD diagnoses is not until 4 years. Furthermore, there are disparities in diagnostic practices across socio-economic strata and racial lines. As a result, there is an urgent need for a brief and inexpensive observational tool that is validated against the gold standard ASD diagnostic assessment. We developed the eight-item Autism Mental Status Exam (AMSE), which structures the way ASD signs and symptoms are observed and recorded; initial validation against the ADOS indicates that the AMSE has excellent classification accuracy and inter-rater reliability. Specific aims are to investigate the validity of the AMSE in a community based population of high-risk children ages 30 months to 5 years old. METHODS/STUDY POPULATION: Children suspected of ASD will be administered the AMSE at their community based agency. The same children will then receive an independent comprehensive diagnostic evaluation at the Seaver Autism Center at Mount Sinai School of Medicine. RESULTS/ANTICIPATED RESULTS: Scores on the AMSE will predict with excellent sensitivity and specificity a best estimate clinical diagnosis of ASD. DISCUSSION/SIGNIFICANCE OF IMPACT: This research will follow the framework for Practice Based Research set forth in the CTSA's Principals of Community Engagement. The AMSE has the potential to represent the next forefront in community-based ASD diagnosis by facilitating both accurate case ascertainment and appropriate referral to evidence based treatments.
TIBETAN IMMIGRANTS IN NEW YORK: A FOCUS UPON CLINICAL PRACTICES AND PERCEPTIONS AS THEY PERTAIN TO CONCURRENT PATIENT-CARE WITH CONVENTIONAL MEDICAL PRACTICE
Mollik M1,2 1Peoples Integrated Alliance, Dhaka, Bangladesh; 2Prescience Trust Funds, Phoenixville, PA, USA
OBJECTIVES/SPECIFIC AIMS: Tibetan medicines largely depend on individual skills and practical knowledge of the people. In addition over time, different medical traditions have developed in different parts of Tibet and have their own specialties. METHODS/STUDY POPULATION: Since very little is known of the traditional medicinal practices of Tibetan immigrants in New York, the extensive observations were conducted during the period November 2011 to June 2012 and traditional treatment system for the management of different diseases in human which are prevalent in the area were recorded with the help of Tibetan immigrants in the area. RESULTS/ANTICIPATED RESULTS: Tibetan immigrants revealed the medicinal values of different plant parts like leaves, fruits, flowers, seeds, stem-barks, tubers, and roots. They have been using these parts in the form of paste, powder, juice, decoction, infusion, and also in crude form, with other additives like clarified butter, edible-oil, honey, and milk, to relieve from different ailments. Many unknown uses of plants brought to the light by the observations, consisting 36 plants belonging to 24 families to relieve different ailments like skin diseases, jaundice, rheumatism, antidotes, burning micturition, fevers, intestinal worms, menstrual problems, cough, diarrhea, headache, cold, diabetes, toothache, asthma, eye diseases, stomachache, indigestion, piles, cuts, wounds, abscesses, sexual problems, for getting abortion, nasal drops, and to retain pregnancy. DISCUSSION/SIGNIFICANCE OF IMPACT: Information on home-grown use of plants has led to discovery of many medicines in use today. The phytochemical and pharmacological screening, biological assay of the drugs/preparations must be carried out to display the active principles present in these plants.
- Top of page
- SCHOLAR/TRAINEE ABSTRACTS
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
- Research Professionals Abstracts Best Practices
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
HEPATIC DE NOVO LIPOGENESIS IN YOUTH
Santoro N1, Parks EJ2, Pierpont B1, Caprio S1 1Yale University, New Haven, CT, USA; 2University of Texas Southwestern, Dallas, TX, USA
OBJECTIVES/SPECIFIC AIMS: Hepatic de novo lipogenesis (DNL) is a major contributor of hepatic lipid accrual and non-alcoholic fatty liver disease. No studies have been performed in youth. Here, we studied 10 adolescents, mean age 15.8 ± 2.3 and mean BMI 35.8 ± 7.6, who underwent an oral glucose tolerance test (OGTT), fast-MRI to measure hepatic fat content, and assessment of hepatic lipogenesis. METHODS/STUDY POPULATION: Fractional lipogenesis (%DNL) was assessed by challenging the subjects with a drink containing 75 g glucose and 25 g fructose. Subjects were admitted at 5:30 PM of day 1 and were given D2O (3 ml/kg of body water) in three doses. At 7:45 AM on day 2, subjects ingested the drink and underwent hourly blood draws for 6 h. Fasting and sugar-stimulated DNL were expressed as the percentage of VLDL-TG palmitate derived from the lipogenesis (%DNL). Indirect calorimetry was performed in the fasting state (6:00 AM) and one hour after the drink (9:00 AM). RESULTS/ANTICIPATED RESULTS: Fasting%DNL was strongly related to BMI (r = 0.62; P = 0.05) and inversely correlated with both the WBISI (r = –0.66; P = 0.03) and the proportional increase in REE after the drink (r = –0.82; P < 0.004). Interestingly, the change in%DNL after the drink was inversely correlated with BMI (r = –0.75; p = 0.01), and directly correlated with the WBISI (r = 0.63; P = 0.04) indicating that lower body mass and greater insulin sensitivity are associated with appropriate stimulation of lipogenesis after sugar consumption. DISCUSSION/SIGNIFICANCE OF IMPACT: These data are the first in obese children and show that in the fasting state, greater insulin resistance is associated with the highest rates of%DNL. The ability to stimulate lipogenesis after a sugar bolus is a healthy response, as is the ability to increase REE in the fed state. The lack of response in insulin resistance demonstrates metabolic inflexibility at a key stage in development.
AMPLITUDE INTEGRATED EEG (AEEG) ABNORMALITIES AT 24 HOURS AND MRI PREDICT OUTCOME IN HYPOXIC-ISCHEMIC ENCEPHALOPATHY (HIE) WITH THERAPEUTIC HYPOTHERMIA (HT): AN INTERNATIONAL COHORT STUDY
Bonifacio S1, Uria-Avellanal C2, Glass HC1, Huertas-Ceballos A2, Rogers EE2, Gunny R2, Barkovich AJ1, Ferriero DM1, Robertson N2, deVries L3, Groenendaal F3 1UCSF, San Fracisco, CA, USA; 2University College London, London, United Kingdom; 3University Medical Center, Utrecht, Netherlands
OBJECTIVES/SPECIFIC AIMS: The ability to discriminate neonates with HIE at greatest risk of death/disability is important to ensure appropriate follow-up and may help identify those eligible for future therapies. We aim to characterize the outcomes after HT for HIE in an international cohort and determine early predictors of outcome. METHODS/STUDY POPULATION: Neonates with HIE treated with HT in three centers (n = 203) were included. Clinical data, aEEG at initiation and 24 h, and brain MRIs were reviewed. Follow-up occurred at 12–36 mo of age and included Bayley Scales of Infant Development (BSID) III and assessment for cerebral palsy (CP). Multivariate logistic regression was used to determine the factors most strongly associated with death/disability (BSID cog score ≤ 80 or CP). RESULTS/ANTICIPATED RESULTS: Outcome was determined in 180(89%). Mean age at follow-up was 24 mo. Death occurred in 49(27%) & disability in 18(14%). The adjusted odds ratios (OR) for death/disability (95%CI), p-value for each predictive factor are listed: Clinical HIE Factor OR 1.01 (0.4–2.6, 0.9); Severe Initial aEEG OR 1.2 (0.3–4.1), 0.8; Persistent Severe aEEG at 24 h OR 5.5 (1.4–21.4), 0.01; Moderate-Severe Brain Injury OR 7.4 (2.6–20.8), <0.0001. DISCUSSION/SIGNIFICANCE OF IMPACT: In a large international cohort of neonates treated clinically with HT, severe aEEG background abnormalities and brain injury determined by MRI are associated with death/disability. Identification of these factors should prompt clinicians to ensure adequate developmental follow-up and referral for therapeutic services. Early clinical markers and aEEG at initiation of HT were not associated with outcome.
ASSESSING VALIDITY OF STAGING CLASSIFICATION FOR SMALL INTESTINAL NEUROENDOCRINE TUMORS
Kim M1, Warner RR1, Roayaie S1, Harpaz N1, Ward S1, Itzkowitz S1, Wisnivesky J1 1Mount Sinai School of Medicine, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: Small intestinal neuroendocrine tumors (SI-NETs) have a variable clinical course. In this study, we used data from a national population-based cancer registry to validate the ability of a new staging system to predict patient outcomes. METHODS/STUDY POPULATION: We used the SEER database to identify patients with confirmed SI-NETS diagnosed between 1988 and 2009. Information on primary tumor size, depth of tumor invasion, lymph node involvement, and distant metastases was used to assign TNM stage. Kaplan Meier methods and Cox proportional hazards models were used to assess survival differences (up to 10 years from diagnosis). RESULTS/ANTICIPATED RESULTS: We identified 6,792 patients with confirmed SI-NETs and complete stage information. Kaplan Meier analysis showed increasingly worse survival with progressive higher T (T1 to T4) categories (p < 0.0001). Similarly, patients with N0 and M0 disease had improved survival as compared to N1 and M1 disease (p < 0.005). Stage I and stage IIA cancer had comparable survival (10-year survival 97% vs. 94%). 10 year survival for stage IIIB (79%) was better than for stage IIIA (72%). Stage IV cancer had the worst 10-year survival (44%). Adjusted analyses showed that T3 (HR: 3.21, 95% CI: 2.04–5.06) and T4 (HR: 5.33, 95% CI: 3.31–8.58) tumors had worse survival than T1 tumors. T2 had comparable survival (HR 1.14, 95% CI: 0.70–1.85) to T1 tumors. N1 disease was not associated with worse survival (HR: 1.35, 95%CI: 0.86–2.10). DISCUSSION/SIGNIFICANCE OF IMPACT: While the newly proposed staging system for SI-NETs helps to predict prognosis in early and late stage disease, the classification does not discriminate well prognosis among patients with locoregional disease. Involvement of regional lymph nodes (N1 disease) does not appear to add independent prognostic information beyond that provided by T status.
ASSOCIATION BETWEEN PUPILLARY LIGHT REFLEX AND SENSORY BEHAVIOR IN CHILDREN WITH AUTISM SPECTRUM DISORDER
Daluwatte CL1, Miles JH1, Sun J1, Yao G1 1University of Missouri, Columbia, Columbia, MO, USA
OBJECTIVES/SPECIFIC AIMS: Pupillary light reflex (PLR) is a simple noninvasive neurological test that can reveal a rich set of neurological information. Sensory Profile questionnaire is widely used in clinic to evaluate sensory processing in children. Both atypical PLR and sensory processing have been reported in children with autism spectrum disorder (ASD). We investigated here whether the physical PLR measures are correlated with the behavioral sensory measures. METHODS/STUDY POPULATION: We measured PLR and sensory profile in 152 children (10.7 ± 3.4 years old) with ASD and 107 children (10.9 ± 2.9 years old) with typical development (TD) between 6–17 years. PLR was measured in both light and dark adaptations and quantified using 5 parameters: resting pupil diameter, relative constriction, latency, constriction time and redilation time. Participants' parents completed 29 sub-items from the Sensory Profile questionnaire. Associations among PLR parameters and sensory scores were first analyzed using linear correlations. Partial least squares (PLS) regression was then performed to select sensory items which are best associated with PLR parameters. RESULTS/ANTICIPATED RESULTS: A significant correlation was found between PLR constriction and the sensory total score in the ASD group (r ≈ 0.3, p < 0.02) but not in TD group (p > 0.05). PLS regression selected 9 sensory items associated with PLR constriction. Children with ASD who reported “always” on these sensory behaviors had lower PLR constriction than those who reported “never”. DISCUSSION/SIGNIFICANCE OF IMPACT: This is the first study that revealed association between PLR and clinical behavioral assessment. A smaller PLR constriction suggests less parasympathetic modulation. Thus our results implied that abnormal sensory behavior in children with ASD could be associated with lower parasympathetic modulation.
ASSOCIATION BETWEEN URINE CHLOROPHENOL LEVELS, WHEEZING-RELATED PROBLEMS, AND SERUM IGE
Jerschow E1,2, Parikh P2, McGinn AP1, de Vos G1, Jariwala S1,2, Hudes G1,2, Rosenstreich D1,2 1Albert Einstein College of Medicine, Bronx, NY, USA; 2Montefiore Medical Center, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: We tested the association between urine levels of five organic phenols (2,5-dichlorophenol (2,5-D), 2,4-dichlorophenol (2,4-D), orthophenylphenol, 2,4,5-trichlorophenol, and 2,4,6-trichlorophenol), the prevalence of wheezing-related problems, and total serum immunoglobulin E (sIgE), in a US representative sample of people ≥6 years of age in the 2005–2006 National Health and Nutrition Examination Survey (NHANES). METHODS/STUDY POPULATION: The association between urine phenols and sIgE was analyzed in an adjusted linear regression model (after log-transformation of both variables). Associations between phenols and prescriptions for medications for wheezing, wheezing during exercise, and days missed from work or school due to wheezing, were analyzed in logistic regression models separately for atopic and non-atopic participants, with organic phenol levels analyzed in tertiles. RESULTS/ANTICIPATED RESULTS: For testing associations between urine phenols and sIgE, our sample included 2127 NHANES participants. For the analyses of the wheezing-related problems, data was available on 159 atopic and 114 non-atopic participants. We found a positive association between sIgE with 2,5-D and 2,4-D; β-coefficients (p-value) 0.047 (0.03) and 0.056 (0.03), respectively. In atopic participants, 2,5-D was associated with more prescriptions (ORhigh vs. lowest tertile = 2.25, ptrend = 0.02), and with missing work or school (ORhigh vs. lowest tertile = 10.13, ptrend < 0.001), and 2,4-D was associated with missing work or school (ORhigh vs. lowest tertile = 10.68, ptrend < 0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: High urine levels of dichlorophenols are associated with increased sIgE levels and with wheezing-related problems in an atopic U.S. population. The exact reasons underlying these associations remain to be determined.
ASSOCIATION OF METABOLIC ACIDOSIS WITH BONE MINERAL DENSITY IN UNITED STATES ADULTS
Chen W1, Abramowitz MK1 1Albert Einstein College of Medicine, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: Chronic acidosis leads to bone mineral loss by inducing calcium efflux from bone. In this study, we examined the association of serum bicarbonate levels with bone densitometry in subjects from the National Health and Nutrition Examination Survey (NHANES). METHODS/STUDY POPULATION: Our analysis included 11,439 adults (≥20 years old) with estimated glomerular filtration rate (eGFR) ≥15 mL/min who completed dual-energy X-ray absorptiometry (DEXA) testing between 1999 and 2004. We excluded participants who were pregnant or taking bisphosphonates. Bone mineral density (BMD) was measured by DEXA. Using linear regression, serum bicarbonate levels were analyzed as a continuous variable to explore its association with total bone density. RESULTS/ANTICIPATED RESULTS: Participants with lower serum bicarbonate were younger, more likely to be female, non-smoker, had more diuretic use and higher eGFR. There was no statistically significant association between serum bicarbonate levels and total BMD in unadjusted models or while adjusting for age, sex, race, education, smoking status, body mass index, eGFR, calcium, phosphorous, hemoglobin, serum albumin, albuminuria and history of diabetes, hypertension and cardiovascular disease as well as diuretic use in the entire cohort [1.0 mg/cm2 difference in BMD per 1 mEq/L increase in serum bicarbonate with 95% CI (–0.6, 2)]. In sex-stratified analyses, serum bicarbonate level was independently associated with total BMD while adjusting for other covariates among women [2.1 mg/cm2 (0.2, 4.0)] but not among men [1.5 mg/cm2 (–0.4, 3.4)]. DISCUSSION/SIGNIFICANCE OF IMPACT: Lower serum bicarbonate levels were significantly associated with lower total bone density in women, but not men. This suggests a gender difference in the role of acidosis in bone metabolism.
ASYMPTOMATIC RESPIRATORY SYNCYTIAL VIRUS INFECTION IN THE FIRST 6 MONTHS OF LIFE
Marom T1, Ede LC1, Patel JA1, Loeffelholz MJ1, Xiong Y1, Chonmaitree T1 1University of Texas Medical Center, Galveston, TX, USA
OBJECTIVES/SPECIFIC AIMS: Primary respiratory syncytial virus RSV infection is believed to be symptomatic in most cases. Advances in molecular diagnostics have increased RSV detection rate in clinical specimens. Asymptomatic RSV infection rate in young infants is yet unknown. We sought to determine asymptomatic RSV infection incidence in the first 6 months of life. METHODS/STUDY POPULATION: In a prospective cohort of healthy infants followed from near birth, we studied upper respiratory tract infection (URI) incidence and its complications. Nasopharyngeal specimens (NS) were collected at scheduled monthly visits (months 1 to 6), regardless of symptoms, and within 7 days of URI onset.This report includes data from a one-year period (August 2010 to July 2011), for which sequential NS from infants younger than 6 months, collected monthly and during URI, were tested for RSV by PCR assays. RESULTS/ANTICIPATED RESULTS: 454 specimens were collected from 120 infants aged 0–6 months (493 child-years). Of these, 405 samples were available for PC R testing. RSV was detected in 19/405 (5%) from 18 infants: 4/44 (9%) of URI samples and 15/361 (4%) of monthly samples. RSV infection rate was 0.037 per child-month. After categorizing RSV infections according to clinical presentation, 11/18 (61%) infants were symptomatic (all having URI) and 4/18 (22%) also had lower respiratory tract infection, while 7/18 (39%) were asymptomatic. There were no significant demographic and risk factors differences were observed between these two groups, although a higher proportion of asymptomatic RSV cases was observed in the warmer months while symptomatic RSV cases were more frequent in the cooler months. DISCUSSION/SIGNIFICANCE OF IMPACT: RSV infections in the first 6 months of life are asymptomatic in 39%. Asymptomatic RSV infection tends to occur out of the typical “RSV season”.
BAYESIAN META REGRESSION OF CANNABIS FOR CHRONIC NEUROPATHY
Andreae MH1, Sacks H4, Carter G3, Indyk D4, Suslov K4, DiMaggio C2, Hall C1, Johnson M2 1Albert Einstein College of Medicine, New York, NY, USA; 2Columbia University, New York, NY, USA; 3Foundation for Integrative AIDS Research, New York, NY, USA; 4Mount Sinai School of Medicine, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: Chronic peripheral neuropathy (CPN) is frequent and difficult to treat. Smoked cannabis may provide relief, but design and outcomes of available randomized controlled trials (RCT) were too heterogeneous for a traditional meta-analysis. Our Bayesian model converted continuous pain outcomes to dichotomous data, quantifying the number of patients needed to treat (NNT) for a clinically meaningful response. We hypothesized that dose differences may partly explain the between study variance and improve effect estimates and credible intervals. METHODS/STUDY POPULATION: We performed a systematic review and Bayesian random effects meta-analysis of smoked cannabis for CPN. We used a Gibbs sampling scheme in OpenBugs to generate a Monte Carlo sample from the posterior distribution of the model parameters. RESULTS/ANTICIPATED RESULTS: We pooled four RCTs with 132 patients, but encountered variance in design and outcomes. We estimated the odds ratio for benefit as 1.8 with a 95% Bayesian credible interval CI 95% [0.9 to 4.5], NNT 4.3, CI 95% [2 to ∞]. Dose differences partly explained the observed between study heterogeneity in our Bayesian meta-regression (R^2 0.57); the adjusted NNT is 3.42 with CI 95% (1.64, 1520), estimated for a dose of 96 mg. The probability of a positive effect is 98%. The Bayes Factor is 57, signifying strong evidence. DISCUSSION/SIGNIFICANCE OF IMPACT: Smoked cannabis is effective for CPN, improving pain in one patient for every 3.4 patients treated. Our dose-effect meta-regression improved effect estimates and shrank the credible interval. Our innovation is the development of a novel Bayesian meta-regression to partly explain the between study variance in heterogeneous RCTs reporting diverse long term pain outcomes.
CANNABIS USE, SUBSYNDROMAL AND PATHOLOGICAL GAMBLING, AND OTHER PSYCHIATRIC DISORDERS
Hammond CJ1, Pilver CE1, Mayes LC1, Potenza MN1 1Yale University, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Cannabis use and cannabis-related disorders are common in adults and frequently co-occur with subsyndromal and pathological gambling (PG). How cannabis use moderates the relationships between problem-gambling severity and psychiatric disorders is poorly understood. METHODS/STUDY POPULATION: Nationally representative data from the National Epidemiological Survey on Alcohol and Related Conditions (n = 43,093 adults) were examined to investigate how cannabis use moderated the associations between problem-gambling severity (based on PG inclusionary criteria) and Axis-I and Axis-II disorders. Problem-gambling severity groups included low frequency/non-gambling (LFG/NG [reference group]), low-risk gambling (LRG), at-risk gambling (ARG), and problem/PG (PPG). RESULTS/ANTICIPATED RESULTS: Among cannabis users and non-users, greater problem-gambling severity was associated with more psychopathology. Significant cannabis-use-by-problem-gambling-severity interactions across mood, anxiety, substance-use and Axis-II disorders were observed. At the LRG level, interactions were observed for mood disorders, specific phobias, alcohol use disorders, cluster A personality disorders (PDs), and cluster B PD; at ARG level, interactions were observed for panic disorder, non-cannabis substance use disorders, nicotine dependence, and cluster A PDs; at the PPG level, interactions were observed for major depression, and cluster A and B PDs. In all cases, associations between problem-gambling severity and psychopathology were stronger among cannabis non-users compared to cannabis users. DISCUSSION/SIGNIFICANCE OF IMPACT: Cannabis use moderates the relationships between problem-gambling severity and psychiatric disorders. Understanding these relationships holds potential for developing improved interventions. Support: NIDA, NIMH, NCRG.
CATHETER-ASSOCIATED URINARY TRACT INFECTIONS IN UNITED STATES HOSPITALS: 2001 TO 2010
Daniels KR1,2, Lee GC1,2, Frei CR1,2 1University of Texas at Austin, San Antonio, TX, USA; 2University of Texas Health Science Center at San Antonio, San Antonio, TX, USA
OBJECTIVES/SPECIFIC AIMS: Catheter-associated urinary tract infections (CAUTIs) have prompted clinical and political prevention initiatives in the United States. This study aims to determine if these strategies are associated with lesser incidence or better treatment outcomes for patients with these infections. METHODS/STUDY POPULATION: This was a retrospective analysis of the U.S. National Hospital Discharge Surveys from 2001 to 2010. Eligible cases included adults at least 18 years of age with an ICD-9-CM diagnosis code for urinary tract infection and a procedure code for either urinary catheter placement or other major procedure as defined by the Healthcare Cost and Utilization Project. Data weights were used to derive national estimates. CAUTI rates were presented as infections per 100 catheterizations. Finally, multivariable logistic regression was used to identify independent predictors of CAUTIs. RESULTS/ANTICIPATED RESULTS: These data represent 70.4 million discharges of catheterized patients, of which 3.8 million (5.4%) developed a CAUTI. CAUTIs decreased from 9.4 cases/100 catheterizations in 2001 to 5.3 cases/100 catheterizations in 2010. Mortality in CAUTI patients declined from 5.4% in 2001 to 3.7% in 2010. Median (interquartile range) hospital length of stay also declined from 9 (5–16) days in 2001 to 7 (4–12) days in 2010. Independent predictors of CAUTI included female sex, emergency hospital admission, transfer from another healthcare facility, and Medicaid as the principal payment source (p < 0.0001 for all variables). DISCUSSION/SIGNIFICANCE OF IMPACT: The incidence of CAUTIs in U.S. hospitals declined over the study period. Furthermore, patients with these infections experienced lower hospital mortality and shorter durations of hospital stay toward the end of the study period.
CONTRIBUTORS TO DEPRESSIVE SYMPTOMS AMONG OLDER KOREAN IMMIGRANTS WITH TYPE 2 DIABETES
Choi SE1, Reed P2 1Nursing Science, University of California, Irvine, CA, USA; 2Psychology and Social Behavior, University of California, Irvine, CA, USA
OBJECTIVES/SPECIFIC AIMS: To examine 1) prevalence of depressive symptoms among older Korean immigrants with type 2 diabetes; 2) factors associated with depressive symptoms; and 3) whether these relationships with depressive symptoms vary as a function of age or gender. METHODS/STUDY POPULATION: A total of 115 Korean immigrant older adults (age 65 and over), recruited from the community, were assessed for three groups of potential factors that could influence depressive symptoms: demographic factors (age, gender, education, English proficiency), clinical factors (duration of diabetes, co-morbidities, insulin use), and psychosocial factors (general health, diabetes quality of life, family support). Depressive symptoms were measured by the Center for Epidemiological Studies – Depression scale [CES-D]. RESULTS/ANTICIPATED RESULTS: 57% of participants had CES-D scores of 16 or higher. Controlling for other factors, multiple regression showed higher levels of depressive symptoms to be associated with greater impact of diabetes on quality of life (b = 5.15, p = .02) and less family support (b = –4.27, p = .05). The relationship between depression and diabetes impact on quality of life was stronger for men than women (b = –11.36, p < .01). DISCUSSION/SIGNIFICANCE OF IMPACT: The findings suggest that Korean immigrants with type 2 diabetes showed a high prevalence of depressive symptoms, and diabetes related quality of life and family support are important factors to consider in addressing depressive symptoms among this population. Particular attention needs to be paid to men within this population as the relationship between diabetes related quality of life and depressive symptoms is stronger than among women.
DECREASING TIME TO ART INITIATION AND VIROLOGIC SUPPRESSION IN A LARGE CLINICAL COHORT
Haines C1, Fleishman J2, Agwu A1, Gebo K1 1Johns Hopkins School of Medicine, Baltimore MD, MD, USA; 2Agency for Healthcare Research and Quality, Rockville, MD, USA
OBJECTIVES/SPECIFIC AIMS: We evaluated time to ART initiation (AI) and HIV virologic suppression (VS) after the December 2007 DHHS guideline change in subjects enrolled in the HIV Research Network cohort. METHODS/STUDY POPULATION: We reviewed data from all adults at 14 clinics in the HIV Research Network from 2003–10 and defined the guideline eras as 2003–07 and 2008–10. Time to AI and time to VS were defined as time from enrollment to AI and HIV RNA <400 copies/ml, respectively. Cox proportional hazards models were used to estimate the hazard ratio for time to AI and VS from the enrollment date by guideline era RESULTS/ANTICIPATED RESULTS: Among 12,363 new presenters, 75.8% were male, 48.9% Black, 12.4% were IDU with median age of 38 (interquartile range: 30–45 years). Median CD4 at AI was 205 cells/mm3 in 2003–07 and increased to 225 cells/mm3 2008–10 (test for trend p < 0.001). Overall, median time to AI was 21 days in 2003–07 and 14 days in 2008–10 (test for trend p < 0.001) and median time to VS was 137.5 days in 2003–07 and 112 days in 2008–10 (test for trend p < 0.001). Table 1. Multivariable Model of Relative Hazard of Event ART Initiation HIV Suppression Enrollment Year (vs. 2003–7) 2008–10 1.41 (1.35 - 1.47) 1.46 (1.38 - 1.54) CD4 Count at Enrollment (vs. ≥350) 200–349 2.35 (2.22 - 2.48) 1.94 (1.82 - 2.07) 50–199 3.59 (3.39 - 3.81) 2.30 (2.15 - 2.45) <50 3.60 (3.38 - 3.83) 2.31 (2.15 - 2.48) HIV Risk (vs. non IDU) IDU 0.90 (0.85 - 0.96) 0.77 (0.71 - 0.83) *Adjusted for clinic site, age, race/ethnicity, and gender DISCUSSION/SIGNIFICANCE OF IMPACT: Time from enrollment to AI and HIV VS has decreased from 2003–07 to 2008–10, suggesting that both clinical practice and HIV treatment have improved since the revision of the DHHS HIV ART initiation guidelines. Future studies should evaluate changes in clinical outcomes.
DEMOGRAPHIC PREDICTORS OF LONG-TERM WEIGHT LOSS AFTER BARIATRIC SURGERY
Rasmussen-Torvik LJ1, Pacheco JA1, Aufox SA1, Kim KA1, Hungness E1, Smith ME1 1Northwestern University Feinberg School of Medicine, Chicago, IL, USA
OBJECTIVES/SPECIFIC AIMS: We sought to determine predictors of long-term weight loss up to 9.5 years after bariatric surgery using data extracted from electronic health records (EHR) and linear mixed effect models. METHODS/STUDY POPULATION: Participants were drawn from Northwestern's biobank (NUgene). Individuals who had undergone bariatric surgery were identified through billing, surgical history, or procedure codes in the EHR. Surgery dates as and all available weight measurements (with associated dates) were extracted from the EHR. Sex, race/ethnicity, education, marital status, and height were taken from the NUgene intake questionnaire. Linear mixed effects models were implemented in SAS PROC MIXED to model weight loss from 1- 9.5 years post-surgery. Marginal effects of covariates of interest and their interactions with time were included in models to test their effects on weight loss overall and weight change across time. RESULTS/ANTICIPATED RESULTS: 140 individuals in NUgene had undergone Roux-en-Y gastric bypass (RNY) and had at least 1 weight measurement 1 year post-surgery. There were 2125 weight measurements which occurred at least 1 year post-surgery in the dataset; the median weight loss represented by these measurements was 32.6% from pre-surgical weight. The model suggested that individuals typically experienced slight weight regain of about 1% of pre-surgical weight per year after their first year post-surgery. Overall, women, taller individuals, and college graduates experienced significantly (p < .05) less weight loss than men, shorter individuals, and those without college degrees. Higher age and initial weight were associated with significantly (p < .05) slower weight regain after 1 year post-surgery. DISCUSSION/SIGNIFICANCE OF IMPACT: Demographic factors predict overall weight loss and rate of weight regain after 1 year post RNY surgery.
DENTAL CARIES PREVALENCE OF 12-YEAR-OLD CHILDREN IN SAN PEDRO DE MACORÍS, DOMINICAN REPUBLIC
Díaz Nicolás J1, Silva G2, Elías A1, Borrell L3 1University of Puerto Rico, San Juan, USA; 2Universidad Central del Este, San Pedro de Macorís, Dominican Republic; 3Lehman College, City University of New York, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: Dental caries is one of the most prevalent chronic diseases in children. While dental caries is decreasing worldwide in developing countries, the Dominican Republic exhibits the highest (DMFT = 5.8) index in the Caribbean basin in 1997 according to WHO. Moreover, health disparities persist among different socioeconomic groups. However, there is no data available on the prevalence of dental caries across different socioeconomic groups on the Island. The aim of this study is to estimate the prevalence of dental caries to determine the health disparities across different socioeconomic groups and sex in 12-year-old children of San Pedro de Macorís. METHODS/STUDY POPULATION: A cross-sectional study design will be conducted using a multistage probability sample of children aged 12 attending public and private schools in San Pedro de Macorís. Calibrated examiners will perform oral exam following the NIDCR's criteria. Decayed-Missing-Filled Teeth index (DMFT) scores will be calculated to estimate the dental caries prevalence using a generalized linear model. The Significant Caries Index (SiC) will be used to identify the existing gaps of the disease by socioeconomic status and sex. A validated and culturally adapted questionnaire will be employed to collect sociodemographic information. RESULTS/ANTICIPATED RESULTS: Pending DISCUSSION/SIGNIFICANCE OF IMPACT: Identify the groups that disproportionally carry the burden of this disease to design and implement health promotion programs that contribute to reduce the oral health gaps of children residing in San Pedro de Macorís. Partially supported by NIH R25MD007607, NIH S21MDO01830 and Colgate-Palmolive Technology Center. The content is solely the responsibility of the authors.
DEPRESSIVE SYMPTOMS AND SELF-CARE BEHAVIORS IN EMERGING ADULTS WITH TYPE 1 DIABETES
Monaghan M1, McCormick King M1, Cogen FR1, Streisand R1 1Children's National Medical Center, Washington, DC, USA
OBJECTIVES/SPECIFIC AIMS: Emerging adults with type 1 diabetes (T1D) are at increased risk for psychosocial distress and negative health outcomes. Depressive symptoms are directly associated with decreased self-care in adolescents with T1D, but this relationship is less explored in emerging adults with T1D. The goal of this study is to evaluate the associations among depressive symptoms, T1D adherence, and self-care skills associated with independence (executive function, goal setting) in emerging adults. METHODS/STUDY POPULATION: Sixty-three emerging adults with T1D (59% female; 78% Caucasian; M age = 18.58 yrs ± .93; M T1D duration = 8.48 yrs ± 5.12) completed self-report questionnaires on mood, adherence, executive function, and self-care goals. A1c and blood glucose (BG) monitoring frequency were taken from the medical chart. RESULTS/ANTICIPATED RESULTS: Twenty percent of the sample reported elevated depressive symptoms on the CES-D. Greater depressive symptoms were significantly associated with fewer BG checks/day (r = –.30), poorer adherence (r = –.31), poorer executive function (r = .49), and fewer goals for independence (r = –.30; all ps < .05). Depressive symptoms were not associated with A1c. Controlling for gender and ethnicity, depressive symptoms accounted for 7% of the variance in BG monitoring frequency, 11% of the variance in adherence, 25% of the variance in executive function, and 7% of the variance in diabetes goals for independence (all ps < .05). DISCUSSION/SIGNIFICANCE OF IMPACT: A significant portion of emerging adults with T1D reported increased depressive symptoms, and depressive symptoms contributed to self-care skills and goals. Interventions targeting patient adherence during this transitional period should address depressive symptoms, and multicomponent interventions addressing mood and behavior may be the most successful for this age group.
DIAGNOSTIC DISPARITIES OF PEDIATRIC CONGENITAL HEARING LOSS IN APPALACHIA
Bush M1, Bianchi K1, Lester C2, Gal TJ1, Shinn J1, Fardo D3, Schoenberg N4 1University of Kentucky, Lexington, KY, USA; 2Cabinet for Health and Family Services, Louisville, KY, USA; 3University of Kentucky Department of Biostatistics, Lexington, KY, USA; 4University of Kentucky Department of Behavioral Science, Lexington, KY, USA
OBJECTIVES/SPECIFIC AIMS: Congenital hearing loss is a common problem and timely identification and intervention is paramount for the language development. Patients from regions of rural healthcare disparity, such as Appalachia, may have many barriers to timely intervention. The purpose of this study was to examine pediatric congenital hearing loss and the timing of diagnosis in Appalachia. METHODS/STUDY POPULATION: Data from newborn hearing screening in Appalachian and non-Appalachian regions of Kentucky between 2009–2011 was analyzed. We also examined the timing of diagnostic testing, age of diagnosis, and compliance with follow-up appointments in patients with congenital sensorineural hearing loss at our institution. RESULTS/ANTICIPATED RESULTS: In Kentucky during 2009–2011 there were 6,970 newborns that failed hearing screening tests and the prevalence of newborn hearing loss was found to be 1.71 out of 1000 live births (1.28/1000 in Appalachia and 1.87/1000 in non-Appalachia). Failure to obtain appropriate follow-up diagnostic testing was 23.8% in Appalachia and 17.3% in non-Appalachia. Children from Appalachia were found to have a trend toward delayed initial diagnostic testing (p = 0.0557) and were significantly delayed in obtaining a final diagnosis of their hearing loss (p = 0.039) compared with children from non-Appalachian regions. DISCUSSION/SIGNIFICANCE OF IMPACT: Congenital hearing loss in children from Appalachia is a common problem and there is a significant disparity in the age of diagnosis, which has the potential to limit language development. It is vital to further assess the causative factors and develop interventions that can address this hearing healthcare issue.
EARLY TRENDS IN CYSTATIN C AND OUTCOMES IN PATIENTS WITH CIRRHOSIS AND ACUTE KIDNEY INJURY
Belcher J1 1Yale University School of Medicine, for the TRIBE-AKI Consortium, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Acute kidney injury (AKI) is a common and severe complication in patients with cirrhosis. Progression of AKI to a higher stage associates with increased mortality. Intervening early in AKI in patients whose renal dysfunction is worsening may improve outcomes. However, serum creatinine correlates poorly with glomerular filtration in patients with cirrhosis and fluctuations may mask progression early the course of AKI. Cystatin C, a low-molecular weight cysteine proteinase inhibitor, has been proposed as a more accurate marker of glomerular filtration. We sought to compare changes in cystatin C and creatinine levels immediately following onset of AKI as predictors of dialysis and mortality during this early time period. METHODS/STUDY POPULATION: Prospective multi-center cohort study in patients with cirrhosis and AKI. Serum creatinine and cystatin measured daily for 3 days from the onset of AKI. RESULTS/ANTICIPATED RESULTS: Of 106 patients included in the study, 37 (35%) met the composite endpoint. Cystatin demonstrated less variability between samples than creatinine. Patients were stratified into four groups reflecting changes in creatinine and cystatin: both unchanged or decreased 38 (36%) (Scr-/CysC-); only cystatin increased 25 (24%) (Scr-/CysC+); only creatinine increased 15 (14%) (Scr+/CysC-); and both increased 28 (26%) (Scr+/CysC+). With Scr-/CysC- as reference, in both instances where cystatin rose, Scr-/CysC+ and Scr+/CysC+, the primary outcome was significantly more frequent on multivariate analysis, p = 0.02 and 0.03, respectively. However, in the group where only creatinine rose, outcomes were similar to the reference group. DISCUSSION/SIGNIFICANCE OF IMPACT: Changes in cystatin levels early in AKI are more closely associated with eventual dialysis or mortality than creatinine and may allow more rapid identification of patients at risk for adverse outcomes.
EFFECTS OF BETA BLOCKERS ON LEFT VENTRICULAR REMODELING AMONG HISPANIC AND AFRICAN AMERICANS WITH CHRONIC HEART FAILURE
Kelesidis I1, Varughese C1, Hourani P1, Ronald Z1 1Albert Einstein College of Medicine, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: Although beta blockers (BB) have been shown to improve cardiac function, there is individual and ethnic variation in BB clinical response. We examined the effects of BB on left ventricular remodeling among African Americans (AA), Hispanics and Caucasians with systolic heart failure (HF). METHODS/STUDY POPULATION: 185 AA, 159 Hispanics and 74 Caucasians with EF ≤ 40% from any etiology were selected from the HF clinic. Change in left ventricular ejection fraction (LVEF), left ventricular end diastolic dimensions (LVEDD), and degree of mitral regurgitation (MR) in response to 1 year of BB was evaluated retrospectively. LVEF responders to BB were defined as patients with an absolute increase in LVEF ≥5% and LVEDD responders as improvement in LVEDD ≥10% from baseline after one year of BB therapy. RESULTS/ANTICIPATED RESULTS: Overall, there was a significant improvement in LVEF, LVEDD and degree of MR in AA and Caucasians after 1 year of BB (P < 0.001 vs baseline). Compared with other races, Hispanics had no significant improvement in LVEDD and degree of MR and had fewer patients with LVEF response (42.77%), LVEDD response (5.03%) and MR response (16.35%). In multivariable adjusted analysis, Hispanic and AA race were important predictors of LVEF and LVEDD (p < 0.01) but not MR response. DISCUSSION/SIGNIFICANCE OF IMPACT: Although most patients demonstrated improvement of LVEF, there seems to be ethnic variability in the effects of BB on cardiac remodeling. Degree of MR and LVEDD failed to show improvement among Hispanics after one year of BB. An underlying genetic difference might be a potential explanation.
EPIDEMIOLOGY OF COPD AMONG LOW-INCOME BLACK AND WHITE U.S. ADULTS
Aldrich MC1, Munro HM2, Mumma M2, Blot WJ1,2 1Vanderbilt University Medical Center, Nashville, TN, USA; 2International Epidemiology Institute, Rockville, MD, USA
OBJECTIVES/SPECIFIC AIMS: Chronic obstructive pulmonary disease (COPD) is the third leading cause of mortality in the United States. We sought to characterize COPD in a low-income population. METHODS/STUDY POPULATION: Among 70,315 black and white adults age 40–79 in the Southern Community Cohort Study (SCCS) recruited from community health centers throughout the Southeast from March 2002 to September 2009, significantly more whites than blacks and women than men self-reported COPD (white females = 16%, black females = 8%, white males = 13%, black males = 5%) at cohort entry. RESULTS/ANTICIPATED RESULTS: Age at first diagnosis was similar between blacks and whites (41 vs 42 years, respectively), but whites were more likely than blacks with COPD to have ever smoked cigarettes (86% vs 75%). Co-morbidities were significantly greater among COPD participants than those without COPD (notably 40% vs 13% reporting asthma and 13% vs 6% reporting cardiovascular disease). Prevalence of COPD varied across the 12-state Southeastern region, with Kentucky having the highest prevalence (15.0%) and Georgia the lowest (6.6%). Using linkages with the Center for Medicaid & Medicare Services (CMS), we found that only 43% of white and 27% of black SCCS participants with a Medicaid or Medicare encounter listing an ICD9 discharge diagnosis (primary or contributing) of COPD at a date prior to entry into the SCCS, self-reported having COPD in the baseline SCCS questionnaire. DISCUSSION/SIGNIFICANCE OF IMPACT: Assuming our approach to case identification within CMS claims data is sufficiently reliable to remove false positives, these findings suggest that COPD is common, yet underreported, in this low income population and highlight the need for greater surveillance of COPD among medically underserved groups.
FATIGUE AND ITS RELATED FACTORS IN PEOPLE WITH TYPE 2 DIABETES
Singh R1, Kluding P1 1University of Kansas Medical Center, Kansas City, KS, USA
OBJECTIVES/SPECIFIC AIMS: Fatigue is a common complaint among people with type 2 diabetes. Despite this, very little research has been done describing the severity, impact, or cause of fatigue in this population. The purpose of the study was to explore the relationship between fatigue and specific diabetes related factors such as BMI, Glycosylated hemoglobin (HbA1c), sleep, pain, number of complications from diabetes, years since diagnosis, and depression that may be associated with increased levels of fatigue. Secondary aim was to determine the impact of fatigue on quality of life and functional status among individuals with type 2 diabetes. METHODS/STUDY POPULATION: 30 participants with type 2 diabetes, mean age 60.17 ± 6.49 (16 females, 14 males) participated in the study.A cross sectional descriptive design was used. Subjects were administered a set of questionnaires testing fatigue, depression, sleep, pain, number of complications, and quality of life. HbA1c, BMI, and 6 minute walk distance (6MWT) were measured. RESULTS/ANTICIPATED RESULTS: Pearson correlational analysis was performed to determine relationships between fatigue and, potential factors, quality of life and functional status Fatigue was significantly related to BMI (r = .518,P = 0.003), pain (r = .403,P = .027), sleep (r = .403, P = 0.027), depression (r = .395,P = .031), and quality of life (r = –.415,P = 0.023).There was no significant relationship between fatigue and HbA1c, years since diagnosis, or 6MWT. DISCUSSION/SIGNIFICANCE OF IMPACT: Fatigue is a persistent compliant in diabetes and may be related to physiological and psychological factors. The presence of fatigue can undermine diabetes health outcomes and can affect a person's quality of life. Health care providers should thoroughly assess the presence of fatigue in their patients.
FOOT DISORDERS ASSOCIATED WITH FOOT-RELATED SYMPTOMS AND FUNCTION
Golightly YM1, Hannan MT2, Hillstrom HJ3, Jordan JM1 1University of North Carolina, Chapel Hill, NC, USA; 2Hebrew SeniorLife, Boston, MA, USA; 3Hospital for Special Surgery, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: Clinical observations suggest foot disorders are associated with foot pain and poor function. Results of prior research studies of older adults are mixed with some supporting this association and others reporting little to no relationship. The objective of this cross-sectional analysis was to determine whether specific foot disorders were associated with worse symptoms and function in African American (AA) and Caucasian men and women 50+ years old using the 42-item Foot and Ankle Outcome Score (FAOS), a questionnaire examining pain, symptoms, activities of daily living, sport and recreational function, and quality of life. METHODS/STUDY POPULATION: Of the 1670 participants in the Johnston County Osteoarthritis Project with FAOS data, foot disorders data were available for 1658 (mean age 69, mean body mass index [BMI] 32 kg/m2, 68% women, 31% AA). Presence of foot disorders was determined using the validated Foot Assessment Clinical Tool. Associations of each foot disorder with FAOS subscale scores were determined with separate linear regression models, adjusting for age, BMI, sex, and race. Effect modification between each foot disorder and age, BMI, sex, or race was examined (p < 0.10 for interaction was considered statistically significant). RESULTS/ANTICIPATED RESULTS: Hallux valgus, plantar fasciitis, and Morton's neuroma were associated with worse FAOS subscale scores (p-values <0.01 to 0.03). There were no significant associations for hammer toes, overlapping toes, claw toes, or tailor's bunion and no statistically significant interactions. DISCUSSION/SIGNIFICANCE OF IMPACT: Some foot disorders were associated with worse FAOS scores. These findings inform our understanding of how certain foot disorders relate to pain and function, and this knowledge will be used to guide future clinical trials.
GENETIC VARIATIONS IN THE PREPROGHRELIN GENE (GHRL) ARE ASSOCIATED WITH ALCOHOL DRINKING BEHAVIORS
Feduccia AA1, Suchankova PP1, Schwandt ML2, Lee MR1, Oot E1, Bollinger JW1, Lexi AA1, Goldman D2, Hodgkinson C2, George DT2, Hommer DW2, Heilig M1, Ramchandani VA2, Leggio L1 1NIH/NIAAA/NIDA, Bethesda, MD, USA; 2NIH/NIAAA, Bethesda, MD, USA
OBJECTIVES/SPECIFIC AIMS: The orexigenic peptide ghrelin is involved in reward signaling and may enhance the incentive value of food and alcohol rewards. Consistent with preclinical findings, human studies further indicate a role of ghrelin in alcohol dependence. The aim of the present study was to determine if single nucleotide polymorphisms (SNPs) within genes encoding ghrelin, i.e. the preproghrelin gene (GHRL), and the ghrelin receptor (GHS-R1A), i.e. growth hormone secretagogue gene (GHSR), are associated with alcohol consumption and risk of dependence. METHODS/STUDY POPULATION: Data collected from male and female subjects (n = 816) enrolled in the NIAAA research program at the NIH Clinical Center were analyzed with multiple logistic linear regression analysis. The Timeline Followback (TLFB) was used as a self-reported measure of alcohol consumption during the previous ninety days and the Clinical Institute Withdrawal Assessment for Alcohol (CIWA-Ar) scale was utilized as a measure of withdrawal severity during the initial study enrollment. RESULTS/ANTICIPATED RESULTS: Preliminary results indicated a significant association of alcohol intake and number of heavy drinking days with a Leu72Met polymorphism (rs696217) in GHRL. In addition, 2 intronic SNPs (rs35682 and rs35683) were associated with CIWA scores, suggesting a role in alcohol withdrawal. Although none of the SNPs tested for GHSR reached significance for alcohol behaviors, one SNP in the 3' end of the GHSR gene (rs565105) was related to body mass index. DISCUSSION/SIGNIFICANCE OF IMPACT: Findings suggest that variations in GHRL may influence alcohol consumption and provides evidence warranting further investigation into ghrelin signaling system for the development and treatment of alcohol dependence.
GENOME-WIDE ASSOCIATION STUDY OF VANCOMYCIN PHARMACOKINETICS
McGregor TL1, Van Driest SL1, Lu Z1, Vear SI1, Creech CB1, Kannankeril PJ1, Brothers KB1, Potts A1, Bowton EA1, Delaney JT1, Bradford Y1, Wilson S1, Olson LM1, Crawford DC1, Saville BR1, Roden DM1, Denny JC1 1Vanderbilt University Medical Center, Nashville, TN, USA
OBJECTIVES/SPECIFIC AIMS: To identify loci associated with vancomycin pharmacokinetics using genome-wide association. METHODS/STUDY POPULATION: Individuals in BioVU, a de-identified biorepository, treated with vancomycin were included if dose and schedule data, a steady state trough, and European ancestry were present. Those <18 years, on prior dialysis, or undergoing heart transplant were excluded. Vancomycin trough and covariates were extracted from the chart. Ke, the elimination rate constant, was calculated for each individual using dose, schedule, weight and trough. Multiple imputation was used for missing covariates. DNA (N = 781) was genotyped with Illumina Omni1-Quad BeadChips. Association of genotypes to the log-transformed primary outcome (vancomycin trough) and secondary outcome (Ke) were determined using linear regression in PLINK. RESULTS/ANTICIPATED RESULTS: The SNP rs3002142 at 1q41 associated with vancomycin trough (n = 745) with a 25% increase in the geometric mean (beta 0.22, P = 9.5 × 10−9) and Ke (n = 733) with a 14% decrease in the geometric mean (beta -0.16, P = 4.9 × 10−9). A second locus at 5q14.3, rs10085144, associated with an increase in trough (+15%, beta 0.14, P = 6.3 × 10−6) and decrease in Ke (–11%, beta –0.11, P = 4.5 × 10−8). DISCUSSION/SIGNIFICANCE OF IMPACT: Serum levels of vancomycin directly relate to antibacterial efficacy and renal toxicity. Although excreted renally as unchanged drug, interindividual variation in vancomycin levels and Ke is incompletely predicted by clinical factors. This initial study of vancomycin pharmacokinetics presents novel associations of loci at 1q41 and 5q14.3. If validated, this identifies potential mechanisms underlying vancomycin pharmacokinetic variability and may lead to improved outcomes through pharmacogenomics.
HOSPITAL CHARACTERISTICS ASSOCIATED WITH PEDICATRIC CLOSTRIDIUM DIFFICILE INFECTION
Goudie A1,2 1University of Arkansas for Medical Sciences, Little Rock, AR, USA; 2Arkansas Children's Hospital Research Institute, Little Rock, AR, USA
OBJECTIVES/SPECIFIC AIMS: The rate and severity of hospital-acquired Clostridium difficile infection (CDI) in children is increasing. Best-evidence practices exist to reduce the risk of CDI but variations in hospital-acquired CDI rates remain across hospitals. We examine the association between hospital characteristics and the pediatric case-mix adjusted risk of acquiring CDI in hospital. METHODS/STUDY POPULATION: Children (age 1 to 17 years) with a hospital stay of at least 3 days were identified from the 2006 to 2010 Healthcare Cost and Utilization Project (HCUP) National Inpatient Sample discharge records. Children with CDI (cases) were matched to children without CDI (controls) using a high dimensional propensity score obtained from a logistic regression model for the probability of acquiring CDI that included demographic as well as classifications for 193 clinical diagnoses and 161 procedure covariates. A matched case-control conditional logistic regression analysis including patient-level demographics and hospital characteristics was performed. RESULTS/ANTICIPATED RESULTS: Compared to children in hospitals with the lowest quartile of bed days paid by private insurers, children in hospitals in the third and fourth highest quartiles had 31% higher odds of acquiring CDI (p = 0.02). There were no differences in the odds of acquiring CDI across hospitals that were delineated by the percentage of Medicaid paid discharge quartiles or children's hospital. Compared to children with stays in large bed size hospitals (HCUP definition), those in small bed size hospitals were 25% less likely to acquire CDI (p = 0.0002). DISCUSSION/SIGNIFICANCE OF IMPACT: After matching children for the adjusted risk of acquiring CDI, those with stays in hospitals with higher than median costs paid by private insurers or seen in a large hospital were most at risk of acquiring CDI.
HPV GENOTYPES AND RISK FACTORS ASSOCIATED WITH PENILE CANCER IN PUERTO RICO
Perez-Ruiz CM1,3, Martinez-Ferrer M2, Puras-Baez A3 1University of Puerto Rico - Medical Sciences Campus, San Juan, USA; 2University of Puerto Rico - Comprehensive Cancer Center, San Juan, USA; 3University of Puerto Rico - School of Medicine, Urology Section, San Juan, USA
OBJECTIVES/SPECIFIC AIMS: The incidence of penile cancer is four times higher in Puerto Ricans as compared with other racial and ethnic groups in the mainland of the United States. HPV has been associated to play an important role in the development of penile cancer. The objective of this study is to determine the prevalence of HPV genotypes 16 and 18 in penile cancer and correlate with clinicopathologic characteristics in Puerto Ricans. METHODS/STUDY POPULATION: Tissue and samples will be collected from patients diagnosed with penile cancer (ICD-9 187) and that had surgical treatment in the University of Puerto Rico, School of Medicine, Urology Clinics. Patients will be identified and will after surgical intervention we will acquire penile cancer and normal adjacent tissue for evaluation at UPR Comprehensive Cancer Center. We will subject this sample on PCR amplification and study for specific HPV genotypes. Also patients prior to intervention will be administered a consent and questionnaire to assess the clinicopathologic risk factors. RESULTS/ANTICIPATED RESULTS: We anticipate that penile cancer in Puerto Ricans will be associated mostly with HPV 16 genotype in addition to circumcision practice, hygienic standard, and phimosis. DISCUSSION/SIGNIFICANCE OF IMPACT: This will be the first study that will determine the HPV genotypes, the clinical and behavioral data in Puerto Ricans. This knowledge will be useful to promote health policies, preventive measures, and to further understand the benefits of male vaccination. This project is partially supported by The NIH-NGA R25MD007607 NIMHHD
IMPACT OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE SEVERITY ON INCIDENT COGNITIVE IMPAIRMENT IN OLDER ADULTS
Chang SS1, Leo-Summers L1, Lee HB2, Vaz Fragoso CA1 1Yale School of Medicine, Dept. of Internal Medicine/Geriatrics, New Haven, CT, USA; 2Yale School of Medicine, Dept. of Psychiatry, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Cognitive impairment increases in prevalence with aging and commonly coexists with severe chronic obstructive pulmonary disease (COPD). Yet, little is known about the extent to which COPD severity contributes to the risk of cognitive impairment in older adults. METHODS/STUDY POPULATION: Data were drawn from the Cardiovascular Health Study, a prospective multi-center cohort of cognitively-intact U.S. community-dwelling adults aged ≥65 with COPD (N = 635). Follow-up began at baseline and continued for 3 years. COPD was established by spirometric criteria for airflow limitation, using the Lambda-Mu-Sigma (LMS) method, an approach which accounts for age-related changes in pulmonary function. Using proportional hazards regression, we evaluated the relationship between COPD severity, staged according to LMS-derived spirometric Z scores (mild, moderate, and severe), and incident cognitive impairment, ascertained by a Modified Mini-Mental State score ≤80 or a Digit Symbol Substitution Test score <19. Competing risk-Cox analyses were performed which treated the occurrence of death as a competing event. Models were adjusted for age, race, gender, education, smoking, and other comorbidities. RESULTS/ANTICIPATED RESULTS: Baseline prevalence of mild, moderate, and severe COPD was 33.2%, 32.4%, and 34.3%, respectively. Of those who became cognitively impaired (N = 96), 25% had mild, 33.3% had moderate, and 41.6% had severe COPD. Worsening levels of COPD severity were associated with a higher risk of cognitive impairment (adjusted HR = 1.45, 95%CI = 1.13–1.87). DISCUSSION/SIGNIFICANCE OF IMPACT: The risk of cognitive impairment increases with greater COPD severity. Evaluation of older adults with COPD should routinely include cognitive screening.
INDUCED SPUTUM BIOMARKERS, TRAFFIC POLLUTION, AND ASTHMA CONTROL IN OLDER ADULTS
Epstein TG1, Kesavalu B1, Ryan P2, Bernstein J1, Villareal M1, Bernstein D1 1University of Cincinnati, Cincinnati, OH, USA; 2Cincinnati Children's Hospital, Cincinnati, OH, USA
OBJECTIVES/SPECIFIC AIMS: Traffic pollution exposure greatly influences asthma control in adults >65 years. We will define the relationship between exposure to traffic pollutants, induced sputum cytology and biomarkers, including myeloperoxidase (MPO), and eosinophil peroxidase (EPO), and asthma control in older adults. METHODS/STUDY POPULATION: Twenty non-smoking subjects were recruited from a KL2-supported cohort of 176 asthmatics >65 years. Elemental carbon attributable to traffic (ECAT), a surrogate for diesel exposure, was determined by land use regression. Five subjects with Asthma Control Questionnaire (ACQ) scores > 1 and ECAT > 0.39 μg/m3, 5 subjects with ACQ > 1 and ECAT < 0.39, 5 subjects with ACQ < 1 and ECAT > 0.39, and 5 with ACQ < 1 and ECAT < 0.39 were selected. Sputum differential cell counts were performed by 3 independent readers. MPO and EPO were determined by ELISA and a functional assay. Log transformed cell counts and MPO/EPO were compared with ACQ scores and ECAT. RESULTS/ANTICIPATED RESULTS: Higher eosinophil cell counts were significantly associated with poorer ACQ scores (mean = 15.3% eosinophils among uncontrolled asthmatics versus 6.3% among controlled asthmatics; p = 0.05). There was no relationship between neutrophil counts and ACQ scores. Higher MPO was significantly associated with high ECAT (p < 0.01 and β = 4.0) and trended towards an association with poorer ACQ (p = 0.09). Higher EPO was significantly associated with high ECAT (p < 0.05) and poorer ACQ (p < 0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: Eosinophilic inflammation is significantly associated with poorer asthma control in older asthmatics. Exposure to traffic pollutants may impact asthma control in older asthmatics through either or both eosinophilic and neutrophilic inflammation. Studies including age-matched healthy controls and an expanded sample size are on-going.
INSIGHTS INTO THE EVOLVING IMPACT OF MULTIDRUG RESISTANT ORGANISMS (MDROS) THROUGH THE INTEGRATION OF MOLECULAR AND EPIDEMIOLOGIC APPROACHES
Perez F1,2, Bonomo RA1,2 1Louis Stokes Cleveland VA Medical Center, Cleveland, OH, USA; 2Case Western Reserve University School of Medicine, Cleveland, OH, USA
OBJECTIVES/SPECIFIC AIMS: To explore the convergence of resistance, virulence and transmission in key MDROs through the investigation of: 1) temporal changes in the molecular epidemiology of A. baumannii; 2) molecular signatures of K. pneumoniae associated with infection and mortality; and, 3) clonal lineages of P. aeruginosa affecting vulnerable patients. METHODS/STUDY POPULATION: Surveillance of MDROs in Northeast Ohio integrating studies of risk factors and clinical outcomes in hospitalized patients with antibiotic resistance detection, genotyping, and whole genome sequencing. RESULTS/ANTICIPATED RESULTS: Patients with MDR and carbapenem-resistant A. baumannii, K. pneumoniae and P. aeruginosa experience high mortality, possess multiple co-morbidities and frequently reside in long-term care facilities. Since 2007, A. baumannii belonging to European clone 2 and harboring carbapenemases OXA-23 and OXA-24 displaced a pre-existent carbapenem-susceptible strain in Northeast Ohio. Among sequence type (ST 258) carbapenem-resistant K. pneumoniae, two different genotypes containing KPC-2 or KPC-3 have emerged, and genome sequencing illuminates distinct host pathogen factors. An international clone (ST 233) of carbapenem-resistant P. aeruginosa producing VIM-2 metallo-β-lactamase has spread into community hospitals in Northeast Ohio. DISCUSSION/SIGNIFICANCE OF IMPACT: MDROs are a serious threat to hospitalized patients, given the lack of safe and reliable treatment options and challenges in the development of new antibiotics. The emergence of successful clonal lineages and the introduction of novel carbapenemase genes may signify a major impact on clinical outcomes. Continued integrated surveillance is needed in order to create interventions that improve the diagnosis, treatment and control of MDROs.
LOW LEVELS OF SERUM VITAMIN D AND PERIODONTAL DISEASE IN A SELECTED GROUP OF PUERTO RICAN ELDERLY
Abreu Fuerte OJ1, Elias Boneta AR1, Hernandez R1, Pousa MS1, Gonzalez RF1, Rodriguez RJ1, Garcia Rivera EJ1, Tatakis DN2, Palacios C1 1University of Puerto Rico - Medical Sciences Campus, San Juan, USA; 2Ohio State University - School of Dentistry, Columbus, OH, USA
OBJECTIVES/SPECIFIC AIMS: 1a: Evaluate the levels of serum vitamin D in patients 60 years older from the dental service in the University of Puerto Rico Hospital “Dr. Federico Trilla” in Carolina. 1b: Evaluate the prevalence of periodontitis in our study population. 1c: Evaluate the relationship between serum vitamin D levels and periodontal disease in our study population. METHODS/STUDY POPULATION: Data collection will be obtained through a cross-sectional study with a sample population of 40 patients 60 years older. Periodontal exams will be conducted using the General Practice Residency in Dentistry facilities at the University of Puerto Rico Hospital “Dr. Federico Trilla”. Periodontal charts will be completed and intravenous blood samples will be collected in order to evaluate the serum vitamin D level of each participant subject. RESULTS/ANTICIPATED RESULTS: We anticipate that low serum vitamin D levels are associated with periodontal disease among our study population of Puerto Ricans 60 years older. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings could lead to perform future island-wide studies in support of the oral health of the Puerto Rican population. The study results may support screenings of at risk populations for low serum vitamin D levels and possibly the supplementation of such patients with vitamin D, in order to foster periodontal disease prevention and effective treatment in Puerto Ricans. “This work was partially supported by the National Institute on Minority Health And Health Disparities of the National Institutes of Health under Award Number R25MD007607. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.”
MATERNAL DEPRESSION CARE-SEEKING INITIATED IN PEDIATRIC SETTINGS
Fernandez y Garcia E1, Paterniti D1, Kravitz R1 1University of California Davis, Sacramento, CA, USA
OBJECTIVES/SPECIFIC AIMS: Few studies examine pediatrician-recommended maternal depression care-seeking beyond the postpartum period. We sought to examine depression care-seeking narratives of mothers asked to seek care for a pediatric-administered positive depression screen. METHODS/STUDY POPULATION: Depression screen positive mothers (N = 104) presenting April-October 2011 with children for well-child care were enrolled in a pilot RCT (ClinicalTrials.gov) of a novel intervention to motivate mothers to seek depression care. Two weeks post intervention, an RA, blinded to group participation, assessed outcomes by phone survey, ending with: “Tell us, in your own words, how and why you decided to try to contact/not contact someone about your possible low mood or depression.” Qualitative responses were transcribed verbatim. Two investigators analyzed each response statement iteratively. Multiple statements by the same respondent were assessed individually. Statements were divided into “care-seeking” and “not care-seeking,” then grouped thematically within each category. Discrepancies in coding were discussed, and consensus achieved based on clinical relevance and potential application. RESULTS/ANTICIPATED RESULTS: Of available mothers at 2 weeks (N = 85), 45% had children over 6 months of age (beyond postpartum) and 51% were in the intervention group. Reasons for not care-seeking included: 1) No Perceived Need; 2) Treatment Ambivalence; and 3) Internal and Systems Barriers. Reasons for care-seeking included: 1) Needed Help Coping with Stressors and 2) Avoiding Effects of Depression. DISCUSSION/SIGNIFICANCE OF IMPACT: Depressed mothers who are beyond postpartum and identified in pediatric settings may experience specific barriers and facilitators to depression care-seeking that should be incorporated into effective pediatric-based maternal depression interventions.
METHYLATION OF THE BMP/TGFa PATHWAY AND CORRELATION WITH SURVIVAL IN CHILDHOOD GERM CELL TUMORS
Musselman J1, Poynter J1,2, Ross J1,2 1University of Minnesota, Minneapolis, MN, USA; 2Masonic Cancer Research Center, Minneapolis, MN, USA
OBJECTIVES/SPECIFIC AIMS: Childhood germ cell tumors (GCTs) are a rare, heterogeneous group of tumors classified together because of their common origin in the primordial germ cell. Previous studies have identified epigenetic alterations in certain histologic subtypes of GCT, especially in genes in the BMP/TGF beta pathway. Interestingly, an analysis of GCT cell lines suggested that DNA methylation was related to resistance to cisplatin, the main chemotherapeutic agent used in these tumors. In this study, we will evaluate the impact of methylation patterns on disease progression and survival. METHODS/STUDY POPULATION: DNA will be extracted from tumor samples collected from 162 children with a diagnosis of GCT who were previously enrolled in an epidemiologic study. We will use pyrosequencing to quantify methylation at each of 20 CpG loci in the BMP/TGF beta pathway and in repeated LINE-1 elements as a measure of global DNA methylation. General linear regression and Kaplan-Meier methods will be used to determine whether methylation differs with respect to tumor histology and survival. RESULTS/ANTICIPATED RESULTS: We hypothesize that increased methylation of gene promoters and global hypomethylation as measured by LINE-1 will be found in GCTs, with a greater degree of aberrant methylation patterns associated with increased tumor aggression, resistance to treatment and poorer survival. DISCUSSION/SIGNIFICANCE OF IMPACT: This will be the first study to evaluate the association between DNA methylation and outcomes in GCTs. The results would have significant translational implications if DNA methylation predicts cisplatin resistance or other outcomes. The results of this project could improve treatment decisions and possibly identify alternative treatments in cases which currently have poorer prognosis or resistance to standard therapies.
MISSED OPPORTUNITIES FOR DEPRESSION SCREENING IN PATIENTS WITH ARTHRITIS IN THE U.S
Margaretten M1, Katz P1, Schmajuk G1, Yelin E1 1UCSF, San Francisco, CA, USA
OBJECTIVES/SPECIFIC AIMS: Arthritis affects 20% of the adult US population and is associated with comorbid depression. Our objective was to examine national rates of depression and depression screening for patients with arthritis between 2006–2010. METHODS/STUDY POPULATION: We used nationally representative cross-sections of ambulatory patient visits in the US from the National Ambulatory Medical Care Survey from 2006–2010, which included 18,507 visits with a diagnosis of arthritis. When weighted to the US population, this total represents approximately 644 million visits. The unit of measurement was visits where arthritis was diagnosed. Outcomes were survey-weighted estimates of comorbid depression and the prevalence of depression screening among patients with arthritis across patient and physician characteristics. RESULTS/ANTICIPATED RESULTS: Of the 644,419,374 visits related with arthritis, 83,574,127 (13%) were associated with a comorbid diagnosis of current depression. The odds ratio for comorbid depression with arthritis was 1.45 (95% CI 1.34, 1.58). Depression screening occurred at 383,500 (1%) visits associated with arthritis. There was no difference in depression screening rates in patients with arthritis compared to the general population, and there was no difference in screening rates by provider type. Although the prevalence of depression at arthritis visits (13 per 100 visits) was among the highest, the prevalence of depression screening at arthritis visits was the lowest (.68 per 100 visits) compared to visits with other common, chronic conditions. DISCUSSION/SIGNIFICANCE OF IMPACT: Despite the high prevalence of depression with arthritis, screening for depression was done at few arthritis visits, representing missed opportunities to detect a common, serious comorbidity. Improved depression screening by providers would identify susceptible patients and may lead to appropriate interventions.
OBJECTIVELY MEASURED SNORING IS ASSOCIATED WITH CAROTID ARTERY VASA VASORUM DENSITY
Rice T1, Radomski TR1, Mulukutla S1, Aiyer A1, Kip KE2, Reis SE1, Strollo PJ1 1University of Pittsburgh, Pittsburgh, PA, USA; 2University of South Florida, Tampa, FL, USA
OBJECTIVES/SPECIFIC AIMS: Recent studies have suggested that snoring may have its own sequelae and is not just a surrogate for obstructive sleep apnea (OSA). We sought to determine if objectively measured snoring was associated with a greater density of adventitial vasa vasorum, a marker of atherosclerotic plaque vulnerability. METHODS/STUDY POPULATION: 171 adults in a community-based participatory research study of CVD risk, underwent home respiratory polygraphy with airflow, effort and oximetry as well as ultrasonographic measurement of carotid vasa vasorum. Total apneas and hypopneas (required 4% oxygen desaturation) per hour of recording time comprised the apnea-hypopnea index (AHI). A 6% change in frequency oscillation of the airflow signal lasting 0.5 to 3 seconds was scored as a snore. Total snores were indexed to duration of airflow evaluation to yield a snoring index (SI). Microbubble contrast-enhanced carotid ultrasound was used to measure peak videointensity in the arterial adventitia. The maximum vasa vasorum (VVmax) signal was the greater of two sides measured. RESULTS/ANTICIPATED RESULTS: The mean age was 65.3 ± 6.9 years. 72% were women and 61% were Caucasian. There was no relationship between the AHI and VVmax (ρ = 0.09, p = 0.24), but the SI was significantly correlated with the VVmax (ρ = 0.16, p = 0.04). Individuals with an SI above the median SI had a greater mean VVmax, 0.75 (0.09) vs. 0.79 (0.08), compared to those with an SI below the median (p = 0.009). SI was a significant independent predictor (p = 0.046) of VVmax in a linear model that adjusted for age, sex and race. DISCUSSION/SIGNIFICANCE OF IMPACT: This exploratory analysis suggests that snoring, but not OSA, is associated with coronary artery plaque instability. Greater VVmax may represent a possible mechanism that leads to stroke in heavy snorers with or without OSA.
OBSTRUCTIVE SLEEP APNEA AS A RISK FACTOR FOR UNPLANNED ER VISITS POST-BARIATRIC SURGERY
White R1, Nair S1, Shaparin N1 1Albert Einstein College of Medicine, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: Previous research has reported a higher incidence of postoperative complications among patients with Obstructive Sleep Apnea (OSA); however, OSA's role as an independent predictor of complications is equivocal. Our study objective is to determine whether the preoperative diagnosis of OSA is an independent risk factor for post operative complications in patients undergoing bariatric surgery. METHODS/STUDY POPULATION: The study population included all patients who underwent bariatric surgery between January 1, 2006 and June 30, 2011 at Montefiore Medical Center. OSA was defined as being previously diagnosed within 10 years prior to surgery. Demographic characteristics were assessed. Implementation of additional monitoring such as telemetry and transfer to the ICU were compared. Kaplan-Meier analysis was performed for length of hospital stay and emergency room visits post-discharge. RESULTS/ANTICIPATED RESULTS: There were 141 patients with OSA and 583 patients without OSA. The OSA cohort had 2 patients (1.42%) transferred to the ICU and the non-OSA had 2 patients (0.34%) transferred to the ICU. 103 OSA patients (73.05%) were on telemetry and 474 non-OSA patients (81.30%) were on telemetry (p = 0.035). The length of stay was similar. OSA patients had a non-significant increased trend for ER visits at 30 days; a significant rate at 60 days (p = 0.03) with a relative risk of 0.63 (0.45, 0.88) for the non-OSA cohort; and a significant rate at 90 days (p = 0.02) with a relative risk for the non-OSA cohort of 0.61 (0.45, 0.83). DISCUSSION/SIGNIFICANCE OF IMPACT: Patients undergoing bariatric surgery with diagnosed OSA are at a higher risk of 60 day and 90 day postoperative ER visits compared to a non-OSA cohort. Future research will look at specific postoperative complications with a goal of developing a conditional logistic regression model to identify significant risk factors.
OBSTRUCTIVE SLEEP APNEA INCREASES RISK OF INCIDENT DEMENTIA IN COMMUNITY-DWELLING OLDER ADULTS
Ju YS1, Lopez O2, Redline S3, Stein P1 1Washington University School of Medicine, St Louis, MO, USA; 2University of Pittsburgh, Pittsburgh, PA, USA; 3Harvard Medical School, Boston, MA, USA
OBJECTIVES/SPECIFIC AIMS: Obstructive sleep apnea (OSA) is common in dementia and is also associated with cognitive impairment among non-demented individuals. A recent study reported that OSA is a risk factor for incident dementia among elderly women. This study was to determine if obstructive sleep apnea increases risk of dementia in older adults independent of gender. METHODS/STUDY POPULATION: Participants in the Sleep Heart Health Study who were also enrolled in the Cardiovascular Health Study were included. Presence of OSA was detected by ambulatory polysomnography in 1997–1998, and defined as apnea and hypopnea index (with 4% SpO2 desaturations) ≥5/hour. Cognitive status was carefully adjudicated in 1998–1999, as previously described. Only participants with normal cognitive status at this time were included in the analysis. Yearly assessments provided information regarding onset of dementia and vital status. Data were censored at 2006. Time to dementia was compared between those with and without OSA, using Cox regression. Results were adjusted for gender, age, and Apoε4 allele status. RESULTS/ANTICIPATED RESULTS: In this older, community based, cognitively-normal sample (n = 208, mean age 76.9 +3.6 years, 40 male%), OSA was diagnosed in 110 (53%). Dementia developed in 109 (52%) participants. Of the 56 (27%) participants who died by 2006, 25 (12%) had developed dementia and were censored at time of dementia diagnosis. OSA was independently associated with dementia diagnosis (Hazard Ratio (HR) = 1.65, 95% CI = 1.10–2.48, p = 0.017) after adjustment for female gender (HR = 1.57, p = 0.043), age (HR = 1.09, p = 0.002), and Apoeε4 allele (HR = 1.56, p = 0.042). DISCUSSION/SIGNIFICANCE OF IMPACT: OSA increased prospective risk of dementia in an older cohort of men and women independent of age, gender, and Apoeε4 allele status.
PATIENT-REPORTED DYSPHAGIA SYMPTOM SEVERITY AFTER OROTRACHEAL INTUBATION
Brodsky MB1, Gellar JE2, Colantuoni EA2, Dinglas VD3, Palmer JB1, Needham DM1,3 1Department of Physical Medicine and Rehabilitation, Johns Hopkins University, Baltimore, MD, USA; 2Department of Biostatistics, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA; 3Outcomes After Critical Illness and Surgery (OACIS) Group, Division of Pulmonary and Critical Care Medicine Johns Hopkins University, Baltimore, MD, USA
OBJECTIVES/SPECIFIC AIMS: Effects of intubation with ventilation on swallowing are unclear. We hypothesized that the duration of intubation was related to severity of dysphagia symptoms in intensive care unit (ICU) patients. METHODS/STUDY POPULATION: Orotracheally intubated patients receiving ventilation for acute respiratory distress syndrome were recruited from 11 ICUs at 4 hospitals in Baltimore, MD. Patients with acute cerebrovascular disease or pre-existing cognitive impairment were excluded. The Sydney Swallowing Questionnaire (SSQ) was administered at hospital discharge; scores ≥200 are indicative of clinically-important dysphagia. A locally-weighted least squares fit was used to qualitatively assess the relationship of dysphagia and duration of intubation. Logistic regression was then used to evaluate the association of dysphagia with the duration of intubation and 21 other relevant demographic and clinical factors. RESULTS/ANTICIPATED RESULTS: 152 patients were analyzed. The odds of dysphagia increased 1.84 (p = 0.013) times for each additional day of intubation until day 5. After day 5, additional days of intubation did not appear to affect dysphagia (odds ratio [OR] 1.01, p = 0.756). The only other factor independently associated with dysphagia was a pre-existing upper gastrointestinal and/or neurological comorbidity (OR 2.42, p = 0.029). DISCUSSION/SIGNIFICANCE OF IMPACT: Each additional day of orotracheal intubation approximately doubles the odds of patient-reported, clinically-important dysphagia symptoms, for up to 5 days, after which no increase could be detected.
POOR AEROBIC FITNESS MAY CONTRIBUTE TO COGNITIVE DECLINE IN HIV-INFECTED OLDER ADULTS
Shah K1, Hilton T2, Hongmei Y1, Guido JJ1, Luque AE1, Hall WJ1, Dewhurst S1, Mapstone M1 1University of Rochester School of Medicine, Rochester, NY, USA; 2Department of Physical Therapy, Ithaca College, Rochester, NY, USA
OBJECTIVES/SPECIFIC AIMS: To investigate the relationship between cognitive ability and aerobic fitness in HIV-infected older adults (HOA). METHODS/STUDY POPULATION: We conducted a cross-sectional study of HOA on antiretroviral therapy (ART) >50 years of age. Domain specific cognitive function was assessed by means of a neuropsychological battery. Aerobic fitness (VO2peak) was assessed using a graded, progressive treadmill test. RESULTS/ANTICIPATED RESULTS: Thirty-seven HOA on ART (mean ± SD: age 60 ± 6years, BMI 28 ± 5, CD4 663 ± 337 cells/ml, duration since HIV diagnosis 17 ± 7 years; 81% males) completed the cognitive tests. Several domains of cognition were significantly associated with VO2peak by Spearman correlation analysis (p < 0.05). By step-wise adjusted multiple regression, VO2peak remained most frequently and significantly related to many cognitive domains such as verbal and visual memory, visual perception, and language (p < 0.05). We found that participants with higher VO2peak were less likely to have more severe forms of HIV-associated neurocognitive disorders (HAND) such as mild neurocognitive disorder (OR = 0.65; P = .01) and HIV-associated dementia (OR = 0.64; P = .0006). DISCUSSION/SIGNIFICANCE OF IMPACT: In HOA, aerobic fitness is related to cognitive performance on various tasks. The likelihood of cognitive impairment increased with lower fitness levels. Therefore, increased fitness may serve an important factor in maintenance of cognition and neural integrity for aging HIV-infected individuals. Future prospective and large scale studies are needed to evaluate the effect of fitness and vascular stiffness and function on cognition and brain structure among HOA.
PREDICTORS OF RENAL DAMAGE IN SICKLE CELL ANEMIA
Saraf SL1, Zhang X1, Lash J1, Machado R1, Gladwin M3, Little J2, Gordeuk V1 1University of Illinois, Chicago, IL, USA; 2Case Western Reserve Unviersity, Cleveland, OH, USA; 3University of Pittsburgh, Pittsburgh, PA, USA
OBJECTIVES/SPECIFIC AIMS: To identify clinical and laboratory factors related to kidney damage in sickle cell anemia (SCA). METHODS/STUDY POPULATION: We studied 356 adult SCA patients at the University of Illinois (UI) and validated our findings in 413 adult SCA patients from the walk-PHaSST study at steady state. Patients were categorized according to GFR ≥ or < 90 mL/min/1.72 m2, albumin-to-creatinine ratio (macroalbuminuria > 300 mg/g), and hemoglobinuria (urine dipstick positive for heme and microscopy negative for red cells). RESULTS/ANTICIPATED RESULTS: Prevalences in the UI and walk-PHaSST cohorts for GFR <90 mL/min/ 1.73 m2, macroalbuminuria, and hemoglobinuria were 17%, 18%, 36% and 18%, 20%, 24%, respectively. Increasing age and macroalbuminuria were independently associated with GFR < 90 mL/min/1.73 m2 in both cohorts (p < 0.0001). Hemoglobinuria (p < 0.0001) and increasing mean arterial pressure (MAP) (p ≤ 0.016) were independently associated with macroalbuminuria in both cohorts. Increased LDH was significantly associated with hemoglobinuria in both cohorts (p ≤ 0.001). On pathway analysis, the following model had a good fit to the data: 1) LDH associated with hemoglobinuria; 2) MAP and hemoglobinuria associated with macroalbuminuria; 3) age and macroalbuminuria associated with GFR < 90 mL/min/1.73 m2 (RMSEA ≤ 0.046). DISCUSSION/SIGNIFICANCE OF IMPACT: Intravascular hemolysis, hemoglobinuria, elevated MAP and increasing age are associated with kidney injury in SCA. Further research is needed to identify genetic and genomic markers that may put patients at risk for kidney damage and to determine if measures to decrease hemolysis may prevent kidney injury.
PREVALENCE AND OUTCOME OF MULTIPLE LISTING FOR PEDIATRIC HEART TRANSPLANTATION
Feingold B1, Park SY1, Comer DM1, Webber SA2, Bryce CL1 1University of Pittsburgh, Pittsburgh, PA, USA; 2Vanderbilt University, Nashville, TN, USA
OBJECTIVES/SPECIFIC AIMS: Multiple listing (ML) occurs in 6% of renal transplant listings and is associated with shorter waittimes and increased Tx rates. We sought to determine the prevalence and outcome of ML for pediatric heart transplantation (HTx). METHODS/STUDY POPULATION: Data on all pts <18 yrs listed for HTx 4/95–12/09 in the OPTN registry were analyzed. We hypothesized that ML pts would more commonly achieve HTx than single-listed pts. ML was defined as registration at ≥2 centers simultaneously for ≥14d or <14d if the candidate died or was transplanted in that time. Characteristics at listing and waitlist outcomes of propensity score matched single- and ML pts, stratified by listing status and time from first listing to ML, were compared. RESULTS/ANTICIPATED RESULTS: ML occurred in 23 of 6290 listings (0.4%). No candidate was listed at >2 centers simultaneously. Median duration from first to second listing was 35d (0–1015) and median duration of MLs was 32d (3–363). Among ML pts there was a greater proportion of males (78 vs. 56%; p = 0.03). Use of ECMO was less common (0 vs. 11%, p = 0.1) and a prospective crossmatch requirement more common (17 vs. 8%, p = 0.08), though neither reached statistical significance. Urgency status at listing was similar between the groups (1/1A: 61 vs. 64%, 1B/2: 39 vs. 36%; p = 0.45) as were weight, age, diagnosis, use of ventilator or inotropes, public/private insurance, and median income (each p ≥ 0.17). After matching, there was a trend toward increased incidence of HTx for ML pts at 3, 6, and 24 mo (50, 65, 80%) vs. single-listed pts (40, 54, 64%; p = 0.11). DISCUSSION/SIGNIFICANCE OF IMPACT: Multiple listing for pediatric HTx occurs infrequently in the US and may be associated with an increased likelihood of Tx. Improved understanding of factors considered by listing centers and families when contemplating multiple listing is warranted.
PROGRESSION OF LEFT VENTRICULAR DIASTOLIC DYSFUNCTION IN AN URBAN POPULATION
Prendergast HM1, Dudley S2, Kane J3, Richardson D3 1University of Illinois, Chicago, IL, USA; 2university of Illinois-cardiology, Chicago, IL, USA; 3RUSH Medical Center, Chicago, IL, USA
OBJECTIVES/SPECIFIC AIMS: To determine the rate of diastolic dysfunction progression in a predominately ethnic minority population and the associated risk factors. METHODS/STUDY POPULATION: A retrospective cohort study of an echocardiogram database and Electronic Health Record (EHR) of Individuals with echocardiograms showing diastolic dysfunction during a 6-year study period (2006–2012). RESULTS/ANTICIPATED RESULTS: During the 6-year retrospective cohort study period, there were 176 patients in the database with 2 or more echocardiograms demonstrating diastolic dysfunction; representing 701 echocardiograms. The mean time between echocardiograms was 1.37(±0.78) years. The mean age was 64.2 (±10) with 82% female and an average BMI 30.4(±6.5). Demographics were similar across all race/ethnicity groups with slightly younger age and higher percentage of obesity among ethnic minorities. The majority of subjects had Grade I diastolic dysfunction at the initial examination (N = 142 (80.7%)). Approximately 25% (n = 44) of the study cohort demonstrated overall worsening grade of diastolic dysfunction with the majority occurring by year 4. The highest rate of progression was observed at year one. Diastolic dysfunction grade was unchanged in 73% (n = 128), and improved in 2% (n = 4). Being on Angiotensin Converting Enzymes Inhibitors was associated with a lower rate of progression (OR = 0.35, CI (0.17,0.76)) DISCUSSION/SIGNIFICANCE OF IMPACT: Our study showed higher rates of diastolic dysfunction progression in year one in this predominately minority population. This confirms previous studies in nonminority population cohorts demonstrating similar rates of diastolic dysfunction progression. Understanding the role of superimposed cardiovascular risk factors in accelerating progression rates is important to optimize intervention strategies.
RECENT TRENDS IN PATTERNS OF CARE FOR STAGE I NON-SMALL CELL LUNG CANCER
Yu JB1,2, Cramer LD1,2, Soulos PR1,2, Gross CP1,2 1Yale School of Medicine and Yale Center for Clinical Investigation, New Haven, CT, USA; 2Cancer Outcomes, Public Policy, and Effectiveness Research (COPPER) Center at Yale, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Despite the development of non-invasive treatment such as stereotactic radiosurgery (SRS), there remain elderly patients with stage I non-small cell lung cancer (NSCLC) who do not undergo curative treatment. We aimed to measure the proportion of patients undergoing SRS and investigate the regional and individual factors associated with those who did not undergo any curative treatment in 2004–2007. Future analyses will compare the morbidity and mortality of SRS vs. surgery in 2004–2009. METHODS/STUDY POPULATION: The Surveillance, Epidemiology, and End Results-Medicare linked database was queried for all patients aged 67–94 with stage I NSCLC who were enrolled in a Medicare fee-for-service plan in 2004–2007. Yearly treatment percentages were compared using the χ2 test. Instrumental variable analysis is planned to compare complications by treatment type. RESULTS/ANTICIPATED RESULTS: We identified 11,174 patients with stage I NSCLC (mean age 77). In 2004–07, the use of SRS increased from 0.5% to 3.0% (p < .001). The percent of patients receiving no curative treatment decreased non-significantly from 21.3% to 19.8% (p = .29). We anticipate that in 2008–2009, SRS use increased and the percent of patients receiving no curative treatment continued to decline. We anticipate that morbidity from SRS at 1 year is lower than that of surgery and that mortality is equivalent. DISCUSSION/SIGNIFICANCE OF IMPACT: The proportion of patients undergoing SRS is slowly increasing, with no associated decline in patients not receiving curative treatment. The analysis of regional and individual factors associated with the non-receipt of curative treatment and SRS is ongoing, as is the measurement of complications and survival from curative treatment.
RESIDUAL VEIN OBSTRUCTION: A META ANALYSES, IS THE CAUSE OF THROMBOSIS RELEVANT IN ITS UTILITY
Janakiram M1,2, Sullivan MR1, Shcherba M1,2, Guo S1, Billet HH1,2 1Montefiore Medical Center, Bronx, NY, USA; 2Albert Einstein College of Medicine, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: Residual vein obstruction (RVO) at the end of anticoagulation for venous thromboembolism (VTE) has been investigated as a predictive marker for recurrent thrombotic risk .We performed a meta-analysis of published studies to determine whether RVO is associated with recurrent VTE. METHODS/STUDY POPULATION: To assess whether a positive RVO study predicts for recurrent VTE, we performed a meta-analysis of previously published prospective studies between January 1990 and December 2011 in which documented DVTs anticoagulated for a minimum of 6 weeks, had a duplex for RVO were followed for recurrent events. Using STATS Direct meta-analysis software. funnel plots for publication bias, Odds Ratios (OR) and Forrest plots were generated. RESULTS/ANTICIPATED RESULTS: Based on the search criteria and review,13 of the 1955 publications were included. 4008 patients,with 3473 events were included. For all patients with VTE, the presence of RVO was associated with a significantly higher recurrent VTE risk OR 1.93 (1.29 – 2.89). For primary VTE, RVO did not demonstrate a statistically significant increased recurrent VTE risk OR 1.38 (0.87 – 2.08).In secondary VTE, the OR was 2.78 (1.40 – 5.50) but when patients with cancer were eliminated the results were not statistically significant OR 1.73 (0.82 – 3.66). For the two studies with cancer patients, the OR was 5.14 (1.59 – 16.65) for recurrent VTE given a positive RVO study DISCUSSION/SIGNIFICANCE OF IMPACT: We conclude that the presence of RVO is not associated with recurrence in primary or secondary VTE but is highly associated with recurrence in the subset with cancer. Further studies are needed to explore this association and whether a negative RVO in this population may identify a population that can safely have their anticoagulation discontinued.
RESTRICTING BACK PAIN IS ASSOCIATED WITH MOBILITY DISABILITY IN OLDER PERSONS
Makris UE1, Fraenkel L2, Han L2, Leo-Summers L2, Gill TM2 1UT Southwestern Medical Center, Dallas, TX, USA; 2Yale University, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Although back pain is common and costly, few longitudinal studies have evaluated the association between back pain severe enough to restrict activity (restricting back pain (RBP)) and the development of disability in mobility. The objective of this study is to evaluate RBP and subsequent episodes of mobility disability (MD) using a recurrent events framework. METHODS/STUDY POPULATION: We evaluated 709 participants (mean age 77.3y, 63% women) of the Precipitating Events Project, a longitudinal study of community-living persons, aged 70+y, who completed monthly assessments of RBP and MD for up to 159 months. RBP was defined as staying in bed for at least 1/2 day and/or cutting down on one's usual activities due to back pain. MD was defined as needing help with/ inability to complete ≥ 1 of the following tasks in any given month: walking 1/4 mile, climbing a flight of stairs, and lifting/carrying 10 lb. The event rate for MD episodes was estimated using a GEE Poisson model. A recurrent events Cox model was used to evaluate the association between RBP and MD, adjusted for fixed-in-time and time-varying covariates. RESULTS/ANTICIPATED RESULTS: The event rate for MD was 9.4 per 100-person months (95% CI 9.0, 9.8), with a median duration of 2 months per episode. Of the 709 eligible participants, 213 (30%) reported disability in at least 1 of the 3 MD tasks at baseline: 20.2% for walking a 1/4 mile and 19% for lifting/carrying 10 lb. RBP was strongly associated with subsequent MD, with an adjusted HR (95% CI, p-value) of 3.4 (2.9, 3.7, <0.001). Among the 496 participants without MD at baseline the adjusted HR was 3.7 (3.2, 4.3, <0.001). DISCUSSION/SIGNIFICANCE OF IMPACT: In this longitudinal study, RBP was independently associated with MD among older persons. Interventions designed to prevent/decrease RBP may reduce the likelihood of MD.
RISK OF TRAUMATIC INTRACRANIAL HEMORRHAGE IN PATIENTS WITH HEAD INJURY AND PREINJURY WARFARIN OR CLOPIDOGREL USE
Nishijima DK1 1UC Davis School of Medicine, Sacramento, CA, USA
OBJECTIVES/SPECIFIC AIMS: Our objectives for this study were to: 1) identify risk factors for immediate traumatic intracranial hemorrhage (tICH) in patients with mild head trauma and preinjury warfarin or clopidogrel use and, 2) to derive a clinical prediction rule to identify patients at low risk for immediate tICH. METHODS/STUDY POPULATION: A prospective, observational study at two trauma centers and four community hospitals enrolled 982 adult ED patients with mild blunt head trauma (initial ED Glasgow Coma Scale score 13–15) and pre-injury warfarin or clopidogrel use. The primary outcome measure was immediate tICH, defined as the presence of intracranial hemorrhage or contusion on the initial cranial CT. Risk for immediate tICH was analyzed in eleven independent predictor variables. Clinical prediction rules were derived with both binary recursive partitioning and multivariable logistic regression. RESULTS/ANTICIPATED RESULTS: The mean age was 75.4 years. Sixty patients (6.1%) had an immediate tICH. History of vomiting (RR 3.53; 95% CI 1.80–6.94), abnormal mental status (RR 2.85; 95% CI 1.65–4.92), clopidogrel use (versus warfarin) (RR 2.52; 95% CI 1.55–4.10), and headache (RR 1.81; 95% CI 1.11–2.96) were associated with an increased risk for immediate tICH. Both binary recursive partitioning and multivariable logistic regression were unable to derive a clinical prediction model that identified a subset of patients at low risk for immediate tICH. DISCUSSION/SIGNIFICANCE OF IMPACT: While we identified several risk factors for immediate tICH, we were unable to identify a subset of patients who are at low risk for immediate tICH. We recommend urgent and liberal cranial CT imaging in this patient population even in the absence of clinical findings.
SPEECH DISTURBS SCANNING PATTERNS IN INFANTS WHO DEVELOP AUTISM SPECTRUM DISORDERS
Shic F1, Macari S1, Chawarska K1 1Yale University School of Medicine, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Autism spectrum disorders (ASDs) are neurodevelopmental disorders associated with social and communicative difficulties. Several studies have found atypical looking towards people and faces in toddlers with ASD, however it is unknown when these atypical patterns of looking arise. The aims of this study are to examine face scanning in 6-month-old infants who develop ASD in order to understand the developmental nature of atypical face processing and social attention in ASD. METHODS/STUDY POPULATION: 6-month-old infants at high-risk (HR) for ASD and low-risk (LR) controls were shown faces in 3 conditions: Static (neutral face image); Affective (smiling face video); and Speech (talking face video). Eye-tracking tracked where participants looked. At 24- or 36-months of age, infants were classified into: ASD (N = 12), atypically developing HR (HR-ATYP; N = 29), typically developing HR (HR-TD), or low risk typically developing (LR-TD). RESULTS/ANTICIPATED RESULTS: 6-month old infants who developed ASD showed generally decreased attention to all scenes with faces, and showed decreased looking at socially informative inner areas of the face only in the Speech condition. DISCUSSION/SIGNIFICANCE OF IMPACT: These results suggest that by 6-months of age, infants who develop ASD are experiencing a very different social environment compared to infants who do not develop ASD. General impairments in looking at the scenes in general could signify fundamental attentional impairments or highlight diminished salience for socially salient information. Specific results regarding diminished looking at inner areas of the face only during Speech suggests some quality of speech disturbs face scanning strategies in infants who will develop ASD. Future work will need to isolate the specific mechanisms and nature of these disturbances, as well as their relationship to concurrent and later autism symptoms.
SUN PROTECTIVE BEHAVIORS & BELIEFS AMONG RUNNERS
Daoud A1, Tenforde A1, Fredericson M1, Sainani K1 1Stanford, Palo Alto, CA, USA
OBJECTIVES/SPECIFIC AIMS: Skin cancer is the most common malignancy in the US. This risk is especially relevant in outdoor athletes with high sun exposure. The frequency of sun protective behaviors and beliefs among competitive runners has not been previously evaluated. METHODS/STUDY POPULATION: A survey was solicited to competitive running clubs. Characteristics of the study subjects were summarized using descriptive statistics. Outcome variables of sun protection behaviors were compared across demographic variables, running characteristics and sun protection beliefs using Pearson's Chi-squared test. RESULTS/ANTICIPATED RESULTS: The majority of runners reported that they “sometimes” utilized 5 out of the 7 specific sun protective behaviors asked about. For the remaining 2 sun protective behaviors, the majority said that they did so “almost always.” The primary reason cited by 72% for why they did not use sunscreen while running related to practical concerns. Only 9% cited the use of another sun protective behavior as the reason to forego sunscreen use. 57% of all runners reported that their sun exposure habits were primarily influenced by reasons related to their skin health. Yet, 33% of respondents said that practical reasons or a lack of interest influenced their sun exposure habits most. DISCUSSION/SIGNIFICANCE OF IMPACT: The majority of the runners do not consistently engage in some form of sun protective behavior even though 55% cite either a fear of skin cancer or skin aging as the primary influence of their sun exposure habits. This survey points to comfort, convenience and forgetfulness as reasons for the disconnect between concern about skin health and lack of consistent sun protection. Increased sunscreen use has been seen with simple interventions that increase accessibility. Efforts to educate should not be abandoned, but there may be more area for improvement by shifting efforts to interventions that address issues of convenience and forgetfulness.
SWALLOWING DYSFUNCTION AND BREATHING EFFORT IN PARKINSON'S DISEASE
Troche MS1, Hegland KW1, Okun MS2, Davenport PW3 1University of Florida, Gainesville, FL, USA; 2University of Florida, Department of Neurology, Gainesville, FL, USA; 3University of Florida, Department of Physiological Sciences, Gainesville, FL, USA
OBJECTIVES/SPECIFIC AIMS: The goal of this study was to determine if swallowing-related dyspnea was correlated to the presence of dysphagia or swallowing-related quality of life (SWAL-QOL) in persons with Parkinson's disease METHODS/STUDY POPULATION: Forty patients with moderate PD completed a Swallowing-related Perception of Dyspnea questionnaire during their regular clinical visit. The questionnaire consisted of questions that identified the presence of swallowing-related dyspnea and associated anxiety. Videofluoroscopic swallow study was used to determine the presence or absence of dysphagia. Patient's also completed the SWAL-QOL questionnaire. RESULTS/ANTICIPATED RESULTS: Total SWAL-QOL scores were significantly correlated with both dyspnea and anxiety scores. As SWAL-QOL worsened, dyspnea and anxiety worsened. Additionally, as dyspnea increased as did anxiety. There was no significant correlation between PA score and any other dependent measure. Participants with dyspnea reported significantly more anxiety related to breathing and worse SWAL-QOL as compared to participants without dyspnea (p = .009). DISCUSSION/SIGNIFICANCE OF IMPACT: This study is the first to relate dyspnea, dysphagia, and quality of life in any population. The results revealed that dyspnea and associated anxiety are related to decreased swallowing-related quality of life in this sample of patients with PD. Future studies should compare swallowing-related perceptions of dyspnea and SWAL-QOL scores to respiratory-based measures of swallowing. These studies are necessary given the high incidence of mortality secondary to swallowing disorders and pulmonary sequelae in PD.
THE HOME MICROBIOME AND CHILDHOOD ASTHMA
Ciaccio C1, Kennedy K1, Barnes C1, Portnoy J1, Rosenwasser L1 1Children's Mercy Hospital, Kansas City, MO, USA
OBJECTIVES/SPECIFIC AIMS: Exposure to microorganisms has repeatedly been found to influence the development of atopic diseases, such as asthma. This relationship has been best understood as an inverse correlation between microbial diversity and atopic disease. Innovative techniques, however, have been developed that more comprehensively characterize microbial communities. The aim of this study, therefore, was to characterize the home microbiota of asthmatics and non-asthmatics. METHODS/STUDY POPULATION: This cross-sectional analysis was performed as part of the Kansas City Safe and Healthy Homes Partnership. The microbiome was compared in the dust from homes of asthmatic children (n = 16) and healthy controls (n = 7). Dust samples were collected from home vacuum bags. DNA was extracted and bacterial 16s rRNA genes amplified. Bacterial products were fragmented, biotin labeled, and hybridized to the PhyloChip Array. Phylochip Arrays were scanned using a GeneArray scanner. RESULTS/ANTICIPATED RESULTS: 1746 operational taxonomic units (OTUs) were found in each of the 23 samples. Bacterial genus richness did not differ in the homes of asthmatics and non-asthmatics (p = 0.09). The Bray Curtis distance between samples demonstrated separation between the whole microbiome of the two groups. All of the top 12 OTUs isolated belonged to one of the five phyla, Cyanobacteria, Firmicutes, Actinobacteria, Proteobacteria, and Bacteroidetes and were all increased in abundance in the samples from homes of asthmatic children (p = 0.001 to p = 7.2 × 10−6). DISCUSSION/SIGNIFICANCE OF IMPACT: These results suggest that home dust has a characteristic microbiota which is disturbed in the homes of asthmatics. Further investigations are needed to determine how home microbial exposures influence the respiratory, skin, and gastrointestinal microbiomes of its inhabitants.
THE INAPPROPRIATE DIAGNOSIS AND TREATMENT OF VENTILATOR-ASSOCIATED PNEUMONIA (VAP) IS COMMON IN MEDICAL AND SURGICAL INTENSIVE CARE UNITS
Nussenblatt V1, Avdic E1, Berenholtz S1, Carlson T1, Daugherty E1, Hadhazy E1, Lipsett P1, Maragakis L1, Perl T1, Speck K1, Sandra Swoboda S1, Ziai W1, Cosgrove S1 1Johns Hopkins School of Medicine, Baltimore, MD, USA
OBJECTIVES/SPECIFIC AIMS: The misdiagnosis of VAP leads to inappropriate antibiotic (ABX) due to unnecessary and prolonged therapy. We quantified inappropriate ABX use for VAP and identified risk factors for the continuation of ABX in patients without VAP after 3 days. METHODS/STUDY POPULATION: Patients thought to have VAP were identified in 6 adult ICUs between 2/2009–2/2010. VAP events were reviewed by a committee to determine the appropriateness of the diagnosis and ABX therapy using clinical, microbiologic and radiographic data at diagnosis, day 3 and 8. Outcomes included a) proportion of VAP events inappropriately treated for VAP on days 1 and 3 b) risk factors for continued ABX in patients without VAP at day 3. RESULTS/ANTICIPATED RESULTS: 232 events were identified as VAP by the ICU's. On day 1, 136 (41.4%) events and on day 3, 73 (32%) events were thought to be VAP by the committee. ABX were continued in 155 (77.4%) of patients without VAP on day 3. Having sputum cultured on day 3 was the only risk factor associated with ABX therapy in patients without VAP on day 3 in the multivariate analysis. ABX use for patients without VAP or another infection on 3 led to 1183 excess days of ABX. There were no significant differences in clinical outcomes for patients without VAP or another infection by treatment group. DISCUSSION/SIGNIFICANCE OF IMPACT: The inappropriate diagnosis and treatment of VAP was common and led to 1183 excess days of ABX in patients with no indication for ABX over one year in 6 ICUs. Clinical differences between patients without VAP who had ABX continued or discontinued were minimal, suggesting that clinician behaviors contribute to unnecessary prescribing. Strategies to improve the diagnosis of and ABX use for VAP are needed.
THROMBOPROPHYLAXIS IN CRITICALLY ILL CHILDREN
Faustino ES1 1Yale School of Medicine, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Because of paucity of evidence to guide practice, we hypothesize that significant variability exists in thromboprophylaxis in critically ill children. We aimed to describe thromboprophylaxis practice in critically ill children and to identify patient, physician and pediatric intensive care unit (ICU) characteristics associated with thromboprophylaxis. METHODS/STUDY POPULATION: We conducted a prospective multinational cross-sectional study on thromboprophylaxis in critically ill children on 4 dates spread in a year. We collected data on patient, physician and ICU characteristics, as well as data on pharmacologic (PT) and mechanical thromboprophylaxis (MT). We determined the associations between characteristics and thromboprophylaxis using nonlinear mixed effects model and reported the associations as odds ratio (OR) and 95% confidence interval (CI). Because MT could only be used for children ≥8 years old, that analysis was limited to this age group. RESULTS/ANTICIPATED RESULTS: Of 2,571 children from 59 PICUs in 7 countries, 381 (12.4%) were on PT while 187 (25.3%) of 740 patients ≥8 years old were on MT. Presence of cyanotic congenital heart disease (odds ratio: 7.39, 95% confidence interval: 4.80–11.38, P < 0.001) and spinal cord injury (odds ratio: 15.04, 95% confidence interval: 2.64–85.66, P = 0.002) had the strongest associations with PT and MT, respectively. Adolescence and obesity were both independently associated with PT and MT. Patients from ICUs with >20 beds were more likely to receive PT while those from North American ICUs were less likely to receive PT. Patients from children's hospital were less likely to receive MT. Physician characteristics were not independently associated with PT or MT. DISCUSSION/SIGNIFICANCE OF IMPACT: We identified characteristics associated with thromboprophylaxis in critically ill children. These characteristics are likely important in the decision to provide thromboprophylaxis and may be targeted for future studies.
TRACKING OF BODY MASS INDEX AND PHYSICAL ACTIVITY IN CHILDHOOD: FOLLOW-UP STUDY
Edwards NM1, Khoury PR1, Kalkwarf HJ1, Woo JG1, Daniels SR2 1Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA; 2Children's Colorado, Denver, CO, USA
OBJECTIVES/SPECIFIC AIMS: Evaluate the tracking of body mass index (BMI) and physical activity (PA) from childhood to adolescence. METHODS/STUDY POPULATION: Healthy children enrolled in a longitudinal cohort study had height, weight, and PA (via accelerometry) measured at age 7 (2005–2006). In 2012, we attempted to contact 191 participants now ages 12–15y, regarding height and weight (expressed as BMI z-score (BMIz)) and PA using a validated questionnaire. Spearman rank order correlation was used to evaluate tracking of BMIz and PA from age 7 to follow-up. RESULTS/ANTICIPATED RESULTS: Eighty-nine participant families responded to the survey (response rate = 47%), and 72 had sufficient data at both time points for analysis. BMIz at age 7 was highly correlated (r = 0.79, P < 0.0001) with BMIz at follow up. There was no correlation (r: –0.18–0.16, P > 0.21) between moderate/vigorous physical activity (MVPA) or inactivity at age 7 with any PA measures at follow up. DISCUSSION/SIGNIFICANCE OF IMPACT: From childhood to adolescence, BMI showed a high degree of tracking, which reinforces the importance of prevention for overweight and obesity. PA did not show evidence of tracking and thus likely requires consistent reinforcement to maintain over time. ACKNOWLEDGEMENTS: R01HL064022, KL2TR000078, UL1RR026314, CCHMC HIRC.
TUMOR CHARACTERISTICS IN BREAST CANCER PATIENTS WITH A FAMILY HISTORY
Ricks-Santi LJ1, Ewing A2, Richardson F1, Carter-Nolan P1, Thompson N1, Harrison B1, Spencer C1, Williams C1 1Howard University College of edicine, Washington, DC, USA; 2Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA
OBJECTIVES/SPECIFIC AIMS: The objective of this study was to determine the tumor characteristics of breast cancer patients reporting a family history of cancer. METHODS/STUDY POPULATION: Data on incident breast cancer cases from 2000–2010 was obtained from the Washington, DC Cancer registry. Variables such as estrogen, progesterone, and Human Epidermal Growth Factor 2 receptor status, as well as stage and grade were analyzed using regression and chi-square analyses in those that self-reported with a family history of cancer. Additionally, the molecular subtypes were compared when ER, PR, and HER2 status were available. RESULTS/ANTICIPATED RESULTS: Family history was associated with having ER and PR negative breast cancer (OR = 1.26 95% CI: 1.02–1.57; p = 0.029), male breast cancer (OR = 2.11 95% CI: 1.37–3.90; p = 0.017), and breast cancer diagnosed under the age of 50 (OR = 1.27 95% CI: 1.09–1.48; p = 0.002). Blacks were more likely to have ER or PR negative tumors (OR = 1.51 95% CI: 1.09–2.08; p = 0.008; PR OR for Blacks = 1.46 95% CI: 1.09–1.94; p = 0.028) and were also more likely to have grade 3 tumors (OR = 1.42 95% CI: 1.05–1.93; p < 0.0001). Additionally, blacks were more likely to have ER/PR negative tumors (OR = 1.5 95% CI: 1.088–2.064; p = 0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: Remarkably, our data shows that the tumor characteristics of hereditary and familial breast cancer are more highly pronounced in blacks compared to whites and Hispanics. Therefore, a greater effort to collect family history and to provide genetic services, such as genetic counseling and genetic testing, to identify individuals at risk for hereditary or familial cancer must be undertaken in African American communities.
UNDERSTANDING RHEUMATOID-ARTHRITIS ASSOCIATED INTERSTITIAL LUNG DISEASE
Lee JS1 1UCSF, San Francsico, CA, USA
OBJECTIVES/SPECIFIC AIMS: Rheumatoid-arthritis associated interstitial lung disease (RA-ILD) is a well-described extra-articular manifestation of RA affecting increasing numbers of patients. There are several ILD subtypes described in RA-ILD based on the histopathologic pattern observed on surgical lung biopsy, with usual interstitial pneumonia (UIP) pattern being the most common (RA-UIP) and non-specific interstitial pneumonia pattern being the second most common (RA-non-UIP). Unlike in the idiopathic interstitial pneumonias where lung histopathology is central to disease classification and treatment, histopathologic subtype has not been emphasized in the clinical evaluation of RA-ILD. Our hypothesis is that RA-UIP is a clinically and biologically distinct condition from RA-non-UIP that is defined by a more rapid decline in lung disease and specific molecular markers. METHODS/STUDY POPULATION: This is a two-center prospective cohort study of RA-ILD patients. We will be collecting serial samples of clinical, physiologic, radiologic and biologic data over 4 years. We plan to enroll 188 RA-ILD subjects, with equal numbers of RA-UIP and RA-non-UIP. RESULTS/ANTICIPATED RESULTS: We expect to find that patients with RA-UIP will have more rapid progression, higher mortality, and distinct molecular marker profile compared to RA-non-UIP. DISCUSSION/SIGNIFICANCE OF IMPACT: If our hypothesis is correct, it will transform our understanding and treatment of RA-ILD by demonstrating the importance of disease subtype, and could suggest similarities between RA-UIP and idiopathic pulmonary fibrosis, an idiopathic fibrotic ILD with UIP pattern.
USING GEOGRAPHIC INFORMATION SYSTEMS (GIS) TO IDENTIFY EPIDEMIOLOGY RISKS IN COMMUNITY ASSOCIATED METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS (MRSA) INFECTIONS
Immergluck LH1,2, Chan TB1,2, Malhotra K1, Zhang S1, Jerris RC2 1Morehouse School of Medicine, Atlanta, GA, USA; 2Children's Healthcare of Atlanta, Atlanta, GA, USA
OBJECTIVES/SPECIFIC AIMS: To determine the trends of MRSA (cases) and methicillin sensitive S. aureus (MSSA, controls) infections, identify differences in 'disease clusters' between MRSA cases and MSSA controls and use GIS to look for associations between household crowding and specific proxies of socio economic conditions and MRSA infections. METHODS/STUDY POPULATION: A case control study. Cases (CA-MRSA infections) and controls (community associated methicillin-susceptible Staphylococcus aureus infections) were from 2 hospitals in Atlanta, Georgia. Electronic health records were queried for demographic, and clinical information. Postal addresses were geocoded and area level data was collected from US Census data. Individual and neighborhood level data were linked. Area data on race/ethnicity, poverty, crowding conditions were thematically mapped. RESULTS/ANTICIPATED RESULTS: From 1,029,355 cultures, 6,206 unique patients were identified of which 5,554 had addresses which were geocoded. MRSA infections increased in children from 2002–2006, with CA-MRSA increasing at a more dramatic rate. At the individual level, differences were seen between CA-MRSA and CA-MSSA for race, age, gender and health insurance. Blacks and children living in impoverished areas were more likely to be diagnosed with CA-MRSA infections. CA-MRSA and CA-MSSA showed spatially significant clustering (z > 1.96). DISCUSSION/SIGNIFICANCE OF IMPACT: CA-MRSA infections have continually increased in this population, particularly among black children. Socio-economic factors affect the risk of getting a MRSA infection in children.
VERTEBRAL FRACTURE AMONG BREAST CANCER SURVIVORS IN CHINA
Hsieh E1, Zhang P2, Zhou C2, Zhang R2, Niu X2, Smith J3, Bradley E5, Fraenkel L1, Insogna K4, Qiao Y2 1Yale School of Medicine - Rheumatology, New Haven, CT, USA; 2Cancer Institute and Hospital, Chinese Academy of Medical Sciences, Beijing, China; 3UNC Gillings School of Public Health, Chapel Hill, NC, USA; 4Yale School of Medicine – Endocrinology, New Haven, CT, USA; 5Yale School of Public Health, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: 1. Measure prevalence of vertebral fracture (VF) among a cohort of breast cancer (BrCa) survivors in Beijing, China. 2. Evaluate risk factors for VF among women with BrCa via a self-administered questionnaire and serologic studies of bone-related indices. 3. Measure health beliefs regarding osteoporosis (OP) and OP prevention measures. METHODS/STUDY POPULATION: A random sample of 200 women (age 50–65y) receiving care at CICAMS will be recruited who were diagnosed and initiated on treatment for BrCa prior to 1/1/2008. Study procedures include thoracolumbar x-ray, a self-administered questionnaire, and serum collection. Medical record data will also be collected regarding BrCa diagnosis, treatment history and medical co-morbidities. RESULTS/ANTICIPATED RESULTS: 1. The prevalence of VF in this cohort will be significantly higher compared to the reported prevalence among healthy age-matched women in Beijing. 2. VF risk will be higher among women diagnosed at a later stage and in those with recurrent disease after controlling for age, BMI, smoking or alcohol use, physical activity, and 25-OH vitamin D status. 3. Knowledge regarding OP and OP preventive measures will be limited and subjects will perceive barriers to adopting behavioral change. DISCUSSION/SIGNIFICANCE OF IMPACT: No data exists regarding the prevalence of VF among Chinese BrCa survivors, a population at significant risk for bone disease. Because patients with BrCa may have different attitudes towards their health and perceived risk of illness compared with healthy individuals, it is important to use primary data from this population when formulating interventions.
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ACUTE VS. DELAYED IRON: EFFECT ON RED CELL IRON INCORPORATION IN MALARIA
Cusick S1, Mupere E2, Georgieff M1, Hodges J1, Abrams S3, John C1 1University of Minnesota, Minneapolis, MN, USA; 2Makerere University, Kampala, Uganda; 3Baylor College of Medicine, Houston, TX, USA
OBJECTIVES/SPECIFIC AIMS: Iron deficiency and malaria frequently coexist, threatening the health and development of millions of children. Concurrent iron and antimalarial treatment does not resolve deficiency and may worsen malaria. Staggering the interventions may enhance the efficacy of each intervention. Our aims are: 1) Identify the sequencing of iron and antimalarial treatment that results in the greatest iron incorporation into red blood cells; 2) Determine whether long-term hematological recovery is affected by giving iron immediately or 4 weeks after antimalarial treatment. METHODS/STUDY POPULATION: 100 children 6–36 months with anemia and malaria presenting to Mulago Hospital in Kampala, Uganda, will be treated for malaria and randomized to 4 weeks of iron starting on Day 0 (immediate) or Day 28 (delayed). Children will receive iron stable isotope 57Fe on Day 0 and 58Fe on Day 28 and return for assessment on Day 56. This design permits assessment of whether immediate or delayed iron results in greater red cell iron incorporation at initial supplement administration (Day 0 immediate vs. Day 28 delayed) and impacts hematologic recovery at Day 56. RESULTS/ANTICIPATED RESULTS: Because malaria-associated inflammation impairs iron absorption and metabolism, treating malaria and reducing inflammation before providing iron will result in greater iron incorporation and greater hematologic recovery in children receiving delayed iron. DISCUSSION/SIGNIFICANCE OF IMPACT: This study will establish a physiologically based framework of optimal timing for antimalarial treatment and iron therapy. Future interventions aimed at safely improving iron status of children in malaria-endemic regions can use this framework and be tested in longer-term studies with neurobehavioral outcomes.
AN INVESTIGATIONAL STUDY TO EVALUATE SILDENAFIL FOR THE TREATMENT OF LYMPHATIC MALFORMATIONS
Danial C1, Tichy A1, Tariq U1, Swetman GL1, Khuu P1, Leung T1, Teng J1, Vasanawala SS1, Lane AT1 1Stanford University, Stanford, CA, USA
OBJECTIVES/SPECIFIC AIMS: Lymphatic malformations (LMs) are localized areas of abnormal development of the lymphatic system. Patients may present with pain, secondary infection, complications due to specific organ involvement, and death. Recently, we reported marked regression of LMs in three children after treatment with oral sildenafil. The objective of this investigational open-label study was to assess the efficacy of 20-weeks of oral sildenafil in reducing the volume and symptoms of children with LMs. METHODS/STUDY POPULATION: Dosing for sildenafil was based on the European Medicines Agency guidelines as follows: if the subject weighed more than 20 kg, 20 mg was given three times a day (60 mg/day); if the subject weighed between 8 kg and 20 kg, 10 mg was given three times a day (30 mg/day). LM volume was calculated blindly using magnetic resonance imaging (MRI) that was performed before and after 20 weeks of sildenafil. Clinical response was assessed on weeks 0, 4, 12, and 20 and both the physician and parent evaluated the LM. RESULTS/ANTICIPATED RESULTS: We have complete data on 6 subjects (N = 6; 3 boys, 3 girls; ages 13–79 months). LM locations included head/neck (N = 5) and abdomen (N = 1). Five subjects had a LM volume decrease (0.01–31.70%). One subject had a 4.0% volume increase; however, the subject had improvement in obstructive sleep apnea scores from 8.8 (moderate) with 26 episodes of apnea to 2.9 (mild) with 0 episodes of apnea. Adverse events included nausea, rhinorrhea, mild insomnia, diarrhea, and fever. All LMs softened and became easily compressible. Several subjects requested to continue sildenafil after study completion because of the improvement they perceived. DISCUSSION/SIGNIFICANCE OF IMPACT: A national double-blind, placebo-controlled trial will be necessary to verify the beneficial effects of sildenafil on LMs.
CAN CAPNOGRAPHY IMPROVE PEDIATRIC SEDATION SAFETY IN THE EMERGENCY DEPARTMENT?
Langhan M1 1Yale University, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: To determine if adding capnography to standard monitoring for procedural sedation will increase staff interventions in response to apnea and hypoventilation, such as verbal/physical stimulation or airway repositioning. To determine if this will then decrease the frequency of oxygen desaturations <95%. METHODS/STUDY POPULATION: This was a randomized controlled trial of children age 1–20 years receiving procedural sedation in a pediatric emergency department. Staff were trained on interpretation of capnography. Subjects received both standard monitoring and capnography via nasal-oral cannula, then were randomized as to whether staff saw the capnography monitor (cases) or were blinded (controls). Study personnel recorded vital signs including end-tidal carbon dioxide (ETCO2) levels every 30 seconds and all interventions. Hypoventilation was defined as ETCO2 >50 mmHg or <30 mmHg without crying/hyperventilation. RESULTS/ANTICIPATED RESULTS: Data are presented for 139 eligible subjects, 68 controls and 71 cases. Mean age was 8 years, 59% were male and 57% white with no significant differences between groups. While a similar proportion of cases and controls hypoventilated (46% vs. 45%), a significantly higher rate of events occurred over time among controls (p = .01). Interventions were performed significantly less among cases (OR 0.4, 95% CI 0.2, 0.8), but more often in response to hypoventilation (OR 2.4, 95% CI 1.2, 4.7). When interventions were in response to hypoventilation, the odds of oxygen desaturation were significantly less (OR 0.2, 95% CI 0.1, 0.5). DISCUSSION/SIGNIFICANCE OF IMPACT: Providers with access to capnography provided fewer, but more appropriate interventions for their patients, which led to fewer episodes of hypoventilation and oxygen desaturations. Capnography may increase patient safety during sedation by providing staff with a more sensitive monitor of ventilation.
EFFECT OF AEROBIC EXERCISE INTERVENTION ON PAINFUL DIABETIC NEUROPATHY
Yoo M1, Kluding PM1, D'Silva L1 1University of Kansas, Kansas City, KS, USA
OBJECTIVES/SPECIFIC AIMS: Diabetic peripheral neuropathy (DPN), caused by neuronal apoptosis and inhibition of nerve regeneration, is a common complication of diabetes. Painful DPN is a common phenotype of DPN characterized by prickling, burning, or sharp shooting sensations in the lower limb and foot. The objective of this pilot study is to explore the effect of aerobic exercise on pain in DPN. METHODS/STUDY POPULATION: Sedentary individuals with type 2 diabetes mellitus between ages 40–70 with clinical diagnosis of DPN were enrolled in a 16-week, 3 X week supervised aerobic exercise program. Six subjects (4 males/2 females; age 57 ± 2.76; duration of diabetes 16.5 ± 8.67 years) have completed their intervention as of 1/2013. All subjects are projected to complete their intervention by 5/2013. Brief Pain Inventory-Diabetic Peripheral Neuropathy (BPI-DPN) questionnaire was used to assess pain intensity (worst, least, average, now) and pain interference with daily life (activity, mood, walk, normal work, relationship, sleep, enjoyment of life) pre and post the exercise intervention. RESULTS/ANTICIPATED RESULTS: Four of 6 (66.7%) participants reported pain due to DPN. In these participants, a significant reduction in average pain score was observed (4 ± 1.15 pre to 2 ± 1.15 post, p = 0.046), while there was no change in scores for worst, least, and current level of pain. A significant reduction in average pain interference score (4.68 ± 3.13 pre to 1.06 ± 0.95 post, P = 0.035) was observed following the intervention. DISCUSSION/SIGNIFICANCE OF IMPACT: These preliminary results show a reduction in average pain intensity and perceived interference in daily life by painful neuropathy using aerobic exercise intervention. Results on other participants who complete the intervention will be included in the presentation. Neurotrophic factors will be analyzed as part of this study from a skin biopsy. Further validation by a randomized controlled trial is needed.
EFFECTS OF EXERCISE TRAINING ON AFFECTIVE PAIN REGIONS IN CHRONIC LOW BACK PAIN
Sharma N1, Brooks W1 1University of Kansas Medical Center, Kansas City, KS, USA
OBJECTIVES/SPECIFIC AIMS: Low N-acetylaspartate (NAA), a marker of neuronal integrity, has been found in dorsolateral prefrontal cortex and anterior cingulate cortices in chronic LBP. It is unclear whether standard exercises for LBP can recover NAA concentrations. The objective of this proposal was to determine the effects of exercise training of core strengthening and aerobic fitness on NAA concentration in affective pain regions METHODS/STUDY POPULATION: LBP participants underwent a 12 week-period of core and aerobic exercise training. NAA was quantified in dorsolateral prefrontal and anterior cingulate cortices and a clinical evaluation included measures of function (Modified Oswestry Disability), pain (Short Form McGill Pain Questionnaire, Numeric Rating Scale of pain) and psychological variables (Fear Avoidance Belief Questionnaire, Beck Depression Index) was completed. RESULTS/ANTICIPATED RESULTS: The preliminary results show a trend toward higher concentrations in NAA in both dorsolateral prefrontal cortices and right anterior cingulate following exercise training. We also found significant improvement in clinical measures of function, pain, psychological variables. The magnitude of change (pre-post) in NAA in dorsolateral prefrontal cortex showed trend toward association with change in fear avoidance behavior and depression. A similar trend of correlation with anterior cingulate was not found with small sample. DISCUSSION/SIGNIFICANCE OF IMPACT: Our preliminary results show that long-term exercise induces neuronal changes in areas involved in affective and emotional pain processing that are related to clinical recovery. Although the results need to be confirmed with large studies these findings might broaden our understanding of cellular processes underlying clinical effects of exercise training in chronic LBP.
FAMILY-CENTERED STRATEGIES TO PROMOTE COGNITIVE AND FUNCTIONAL RECOVERY IN HOSPITALIZED OLDER ADULTS
Boltz M1, Galvin JE2 1New York University, New York, NY, USA; 2New York University School of Medicine, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: To test the feasibility and utility of Family Centered Function Focused Care (FC-FFC) to promote functional recovery in acutely ill older medical patients, while improving family caregiver effectiveness and coping. FC-FFC comprises: 1) staff education; 2) family caregiver education to support the role of informed advocate; 3) practices that engage family caregivers in assessment, decision-making and evaluation; and 4) post-acute follow-up. METHODS/STUDY POPULATION: One hundred dyads (hospitalized older adults and family caregivers), 50 from the intervention unit and 50 from two matched comparison units (staff education only) comprise the sample. A repeated measures design will evaluate upon admission, discharge, 14 and 60 days post-discharge: 1) patient outcomes (functional performance/capability, cognition, recidivism) and 2) family carer outcomes (preparation, affect, strain, mutuality). Data also include enrollment and treatment fidelity measures (goal attainment, staff knowledge, adherence). RESULTS/ANTICIPATED RESULTS: Findings will provide evidence for recruitment and engagement feasibility and FC-FFC enactment. A single group t-test will evaluate changes in goal attainment scores in the intervention group. Preliminary effectiveness on patient and family caregiver outcomes will be evaluated using generalized estimating equations (GEE). Independent variables in the models include (1) treatment effects (FC-FFC vs. comparison), (2) time (admission, discharge, 14 and 60 days post discharge); and (3) interaction of treatment with time. DISCUSSION/SIGNIFICANCE OF IMPACT: FC-FFC has the potential to improve outcomes for older adults and family caregivers during and after hospitalization. Reseach supported by Grant UL1 TR000038 National Center for the Advancement of Translational Science, National Institutes of Health.
MEXILETINE FOR THE TREATMENT OF CRAMPS IN ALS
Oskarsson B1, Moore D2, Joyce N1, Macdonald C1, Lomen-Hoerth C3, Mozaffar T5, Rosenfeld J4, Baloh R6, So Y7, Weiss M8, Forshew D2, Katz J2, Miller R2 1UC Davis, Sacramento, CA, USA; 2CPMC, San Francisco, CA, USA; 3UCSF, San Francisco, CA, USA; 4UCSF- Fresno, Fresno, CA, USA; 5UC Irvine, Fresno, CA, USA; 6Cedar Sinai, Los Angeles, CA, USA; 7Stanford, Palo Alto, CA, USA; 8University of Washington, Seattle, WA, USA
OBJECTIVES/SPECIFIC AIMS: Muscle cramps affect most ALS patients, yet there is no consensus on how to treat them. Traditionally quinine was used, but the FDA recommends against its use. Mexiletine blocks persistent Na+ currents there by reducing cramp propensity in motor neurons. It decreases the frequency of muscle cramps by 87% in patients with Machado-Joseph disease. We aim to show that mexiletine reduces cramp frequency in subjects with ALS compared to placebo treatment. METHODS/STUDY POPULATION: A double blind within-subjects crossover design will be employed. After a one week run in, 30 ALS subjects with severe muscle cramps will be randomized to placebo or mexiletine 300 mg/day for their 1st 2 week epoch. After a 1 week washout they will cross into the 2nd epoch for a total study duration of 6 weeks Outcome Measures Daily subject administered count of cramps from day 5–14 of each treatment epoch Daily Cramp and Fasciculation Severity Scores-self-administered 100 point scales Power The power calculation is based on 27 subjects. The primary outcome is the within-subject difference in number of cramps on each treatment. If the true difference between the treatments is on average 1.2 cramps per day (23% reduction from 5.6 to 4.3), we will have 80% power to detect a treatment difference using a two sided 5% significance level. RESULTS/ANTICIPATED RESULTS: We expect to learn whether mexiletine reduces the frequency and/or severity of muscle cramps in ALS. DISCUSSION/SIGNIFICANCE OF IMPACT: If mexiletine is effective in reducing painful muscle cramps this treatment would be immediately available for the large number of ALS patients living in the US and Canada.
MINDFULNESS-BASED STRESS REDUCTION FOR CANCER SURVIVORS WITH PERSISTENT FATIGUE: PILOT STUDY RESULTS AND CURRENT TRIAL METHODS
Johns SA1,2, Brown LF1, Beck-Coon K1, Monahan PO1, Tong Y1, Kroenke K1,3 1Indiana University School of Medicine, Indianapolis, IN, USA; 2Regenstrief Institute, Inc., Indianapolis, IN, USA; 3Center for Implementing Evidence-Based Practice, Roudebush VA Medical Center, Indianapolis, IN, USA
OBJECTIVES/SPECIFIC AIMS: Cancer-related fatigue (CRF) is one of the most prevalent, persistent, and disabling symptoms associated with cancer and its treatment. Acceptable and efficacious interventions for CRF are lacking. This study was designed to examine the effects of Mindfulness-Based Stress Reduction (MBSR) on those with persistent CRF. METHODS/STUDY POPULATION: A heterogeneous sample of 35 cancer survivors (77% breast cancer; mean age 57 years; all had completed chemotherapy and/or radiotherapy > 9 months prior) was randomly assigned to MBSR or wait-list control. The 7-week MBSR course combined training in mindfulness meditation, yoga, and didactic information on stress reactivity. Outcomes, including fatigue interference and severity, were measured immediately post-intervention and at 1- and 6-month follow-up. RESULTS/ANTICIPATED RESULTS: Compared to controls, participants in the MBSR group reported significant decreases in fatigue interference (d = –1.53, p < .0001) and severity (d = -1.55, p < .0001) immediately after the intervention. These results were maintained at 1- and 6-month follow-up. DISCUSSION/SIGNIFICANCE OF IMPACT: Given the results of this pilot study and the high class attendance (90%) and trial retention rates (97%), it seems that patients are interested in and benefit from non-pharmacologic mindfulness-based interventions for CRF. We are currently enrolling 108 fatigued breast and colorectal cancer survivors in a more rigorous trial of MBSR compared to an attention control condition. Findings will guide the development of subsequent trials in CRF management to expand treatment options into community settings.
NO RELATIONSHIP BETWEEN BODY SIZE AND CLINICAL RESPONSE TO INTRAVENOUS HYDROMORPHONE
Xia S1, Birnbaum A1, Choe D1, Hernandez L1, Bijur P1 1Albert Einstein College of Medicine, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: Weight-based dosing of Intravenous (iv) opioid is commonly recommended despite a paucity of evidence. The objective of this study was to examine the association between different body size calculations [total body weight (TBW), body mass index (BMI), ideal body weight (IBW), lean body weight (LBW), body surface area (BSA), adjusted body weight (ABW), predicted normal weight (PNW)] and clinical response to a fixed initial dose of iv hydromorphone. METHODS/STUDY POPULATION: A prospective interventional study was performed. A convenience sample of adult Emergency Department (ED) patients with acute pain (≤ 7 d) deemed to require iv opioids by attending physicians received 1 mg of iv hydromorphone. Pain scores (numerical rating scale, NRS) were recorded at baseline and 30 min after hydromorphone administration. Pain relief, satisfaction, desire for more analgesics, and side effects, as well as patient characteristics (age, gender, race/ethnicity, educational level), pain duration, location, and mechanism were documented. Weight and height were measured. The main outcome measure was change in NRS pain score (baseline to 30 min). Secondary outcomes were NRS ≤ 3, pain relief, satisfaction at 30 min, and desire for more analgesics. Regression analyses were used to examine relationships between body size calculations and analgesic responses. RESULTS/ANTICIPATED RESULTS: 1438 patients were screened and 163 patients were enrolled. Mean age was 39 years, with 62% female, 57.1% Hispanic and 24.5% African American. There were no statistically significant associations between any of the body size calculations and clinical response to iv hydromorphone, after adjusting for confounders (age, gender, race/ethnicity, education level, pain duration, location, and mechanism). DISCUSSION/SIGNIFICANCE OF IMPACT: These results do not support iv opioid dose adjustment based on body size.
PHENYTOIN LEVELS ARE ELEVATED IN CHILDREN RECEIVING THERAPEUTIC HYPOTHERMIA FOLLOWING TRAUMATIC BRAIN INJURY
Empey PE1, Velez de Mendizabal N2, Bell MJ1, Bies RR2, Anderson KB1, Kochanek PM1, Adelson P3, Poloyac SM1 1Univ of Pittsburgh, Pittsburgh, PA, USA; 2Indiana Univ, Indianapolis, IN, USA; 3Phoenix Children's Hospital, Phoenix, AZ, USA
OBJECTIVES/SPECIFIC AIMS: Therapeutic hypothermia (TH) decreases cytochrome p450(CYP)-mediated drug metabolism in vitro and limited clinical data suggest drug levels increase during cooling. Fosphenytoin is metabolized by CYP2C, has a narrow therapeutic range, and is commonly used after traumatic brain injury (TBI). The study objective was to evaluate phenytoin pharmacokinetics (PK) in children receiving this therapy in an RCT for severe TBI. METHODS/STUDY POPULATION: Total and free phenytoin levels were evaluated retrospectively in children with TBI who received 48h of TH (32–33°C) or normothermia. Drug dosing, body temperatures, and demographics were collected during cooling, rewarming, and post-treatment periods (8 days). Nonlinear mixed effects modeling was used to quantify and describe the impact of temperature and clinical covariates on PK. RESULTS/ANTICIPATED RESULTS: A total of 121 total and 114 free phenytoin levels were evaluated in 10 TH- and 9 normothermia-treated children. There was a trend towards elevated free phenytoin levels in the TH group (p = 0.051; 2-way RM ANOVA) to a median of 2.2 mg/L during rewarming that was not explained by dosing differences. Modeling showed TH decreased the time-variant maximum velocity of phenytoin metabolism (Vmax) 4.6-fold (11.6 to 2.53 mg/h) and reduced the overall Vmax by ∼50%. Simulations showed that the increased risk for drug toxicity extends many days after cooling. DISCUSSION/SIGNIFICANCE OF IMPACT: TH significantly reduces phenytoin elimination in children with severe TBI which may lead to increased drug levels for an extended period of time after cooling. PK interactions between hypothermia and medications should be considered when caring for children receiving this therapy. Support: NS052478, TR000146, NS030318.
POTENTIATION OF CETUXIMAB BY INHIBITION OF TREGS IN METASTATIC SQUAMOUS CELL CANCERS OF HEAD AND NECK
Jha GG1 1University of Minnesota, Minneapolis, MN, USA
OBJECTIVES/SPECIFIC AIMS: There are few effective treatment options for patients with metastatic squamous cell cancers of head and neck (HNSCC). The only significant advance in the last few decades has been approval of cetuximab, a monoclonal antibody directed against the overexpressed epidermal growth factor receptor (EGFR). Cetuximab binds EGFR on tumor cells making them targets for natural killer (NK) cell-mediated lysis. The effectiveness of cetuximab is limited by the tumor mediated immune-suppression that inhibits NK and other effector cells. In the current proposal I seek to abrogate this immune-suppression by selectively targeting the inhibitory regulatory T cells (Tregs) using Cytoxan (cyclophosphamide) to assess the potentiation of immunological and clinical response to cetuximab. METHODS/STUDY POPULATION: Eligible patients will receive daily oral cyclophosphamide and weekly cetuximab. Patients will have blood collected for immune monitoring at weeks 0,3,6,12 and biopsies at week 0,6. Restaging CT scans will be done pretreatment and after weeks 6,12. RESULTS/ANTICIPATED RESULTS: The proposed hypothesis will test if Cytoxan can be used to deplete the inhibitory Tregs and improve efficacy of cetuximab, thereby offering an effective and well-tolerated option for patients with locally advanced metastatic HNSCC. The 3 patients enrolled to-date have tolerated Cytoxan without any serious adverse events. DISCUSSION/SIGNIFICANCE OF IMPACT: This will be the first study to demonstrate Treg depletion as a strategy to harness immune response to tumor and potentiate the effects of cetuximab. Information gained from this pilot trial will enable designing a more definitive phase II study. Because immune-deregulation applies to virtually all malignancies, this strategy targeting immune-deregulation will lead to a paradigm shift in how we manage broad range of malignancies.
PREVALENCE OF ASYMPTOMATIC DEEP VENOUS THROMBOSIS AMONG BARIATRIC SURGICAL PATIENTS RECEIVING PERIOPERATIVE ENOXAPARIN VERSUS FONDAPARINUX
Steele KE1, Schweitzer MA1 1The Johns Hopkins University School of Medicine and The Johns Hopkins Center for Bariatric Surgery, Baltimore, MD, USA
OBJECTIVES/SPECIFIC AIMS: To conduct a pilot study using magnetic resonance venography (MRV) to estimate the prevalence of asymptomatic deep venous thrombosis (DVT) in bariatric surgical patients receiving enoxaparin or fondaparinux sodium during their perioperative hospitalization. METHODS/STUDY POPULATION: From July 2010 to July 2012, one hundred twenty-one consecutive bariatric surgical patients from an academic institution were randomized in a double blinded manner to receive either enoxaparin 40 mg twice daily or fondaparinux sodium 5 mg once daily. Two weeks following surgery the patients returned to clinic to undergo magnetic resonance venography to detect the presence or absence of asymptomatic deep venous thrombosis. Main Outcome Measures: The primary outcome was asymptomatic DVT. The secondary outcome was attainment of therapeutic anticoagulation. Safety outcomes were bleeding, symptomatic DVT, or death. RESULTS/ANTICIPATED RESULTS: Two of the 106 patients evaluated developed DVT, one in each arm of the study. 25.5% (p = 0.003) of patients were sub-therapeutic on anticoagulation. Therapeutic levels were associated with fondaparinux use, gastric bypass (vs. sleeve gastrectomy), and elevated preoperative D-dimer levels. No major adverse events occurred in either arm. DISCUSSION/SIGNIFICANCE OF IMPACT: This is an interim analysis of a randomized study for prevention of DVT after bariatric surgery. Both regimens appear to be equivalently effective at reducing the risk of DVT. Further prospective studies are needed to determine the optimal DVT prophylaxis regimen in the bariatric surgical population.
SAFETY OF NIGHTTIME 2-HOUR SUSPENSION OF BASAL INSULIN IN PUMP-TREATED TYPE 1 DIABETES (T1D) EVEN IN THE ABSENCE OF LOW GLUCOSE
Sherr J1, Palau Collazo M1, Carria L1, Steffen A1, Weyman K1, Zgorski M1, Tichy E1, Cengiz E1, Tamborlane W1, Weinzimer S1 1Yale University, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Low glucose suspend feature of the Veo sensor-augmented pump interrupts basal insulin infusion for 2-hrs if patients fail to respond to low alarms. Approval in the U.S. has been delayed given concerns that falsely low glucose levels could lead to ketosis due to suspensions when actual blood glucose (BG) is elevated. METHODS/STUDY POPULATION: We enrolled 17 T1D subjects (age 24 ± 9 yr, duration 14 ± 11 yr, A1c 7.3 ± 0.5) to examine the safety of nighttime 2-hr basal suspension over a range of starting BG levels. Subjects measured blood β-hydroxybutrate (BHB) and BG each night at 9PM and fasting the next morning. On control nights, usual basal rates were continued; on experimental nights, the basal insulin infusion was re-programmed for a 2-hr zero basal rate at random times after 11:30PM (e.g. 1:30–3:30AM or 2–4AM). RESULTS/ANTICIPATED RESULTS: 131 non-suspend nights were compared to 118 suspend nights. BG and BHB were similar at 9 PM in either condition (BG: suspend 144 ± 63 mg/dL vs. non suspend 151 ± 65 mg/dL; p = 0.39 and BHB:suspend 0.09 ± 0.07 mmol/L vs. non-suspend 0.08 ± 0.06 mmol/L; p = 0.47). BG increased by ∼50 mg/dL in the fasting period (suspend 191 ± 68 mg/dL vs. non-suspend 141 ± 75 mg/dL; p < 0.0001) and the frequency of fasting hypoglycemia decreased the morning following suspend nights (suspend 4% vs. non-supsend 21%; p < 0.0001); morning BHB were slightly higher with suspension but clinically equivalent (suspend 0.13 ± 0.14 mmol/L vs. non-suspend 0.09 ± 0.11 mmol/L; p = 0.053). DISCUSSION/SIGNIFICANCE OF IMPACT: Thus, systems that suspend basal insulin for 2-hrs are safe, should not lead to clinically significant ketonemia even if BG is elevated at the time of the suspension, and reduce the incidence of fasting hypoglycemia.
SMOKING DIMINISHES MOTIVATION-REWARD NEURAL RESPONSES TO FOOD CUES INDEPENDENT OF BODY WEIGHT
Jastreboff AM1, Sinha R1, Lacadie C1, Sherwin R1, Potenza MN1 1Yale University, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Co-occurrence of smoking and obesity is synergistically associated with increased risk of cardiovascular disease and cancer. Smoking status is associated with relatively lower body weight, yet why this occurs has not been fully elucidated. Both smoking and obesity have been shown to affect neural responses in reward-motivation pathways. METHODS/STUDY POPULATION: We matched smokers and non-smokers based on BMI, age, and gender and examined whether smoking status altered neural activation in motivation-reward and emotion-regulating brain regions in response to brief guided-imagery of favorite foods. 23 smokers and 23 non-smokers were exposed to trials of personalized food-cue and neutral-relaxing situations using a validated, functional MRI paradigm. RESULTS/ANTICIPATED RESULTS: During exposure to the food-cue condition, smokers as compared to non-smokers exhibited decreased activation in corticolimbic-striatal regions including the caudate, putamen, insula, and thalamus. In condition contrasts of food-cues vs. neutral-relaxing condition, non-smokers exhibited increased activation in the inferior frontal gyrus (IFG), middle temporal gyrus (MTG), putamen, caudate, insula, amygdala, and hippocampus; smokers demonstrated increased activation in the IFG and MTG but not in striatal or limbic regions (p < .05 whole brain corrected). No statistically significant differences were found between smokers and non-smokers with regard to insulin resistance (HOMA-IR), fasting insulin, glucose, leptin, and cortisol levels, or measures of subjective food craving. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings suggest that smokers demonstrate diminished neural responses to food cues irrespective of insulin resistance or BMI status. Differences in neural response to food cues in smokers may contribute to their relatively lower BMI and perhaps to the weight gain seen with attempts to quit smoking.
YOGA AND VIOLENCE SURVIVOR MENTAL HEALTH: A FEASIBILITY STUDY AMONG GROUP-THERAPY CLIENTS
Clark C1, Lewis-Dmello A2, Anders D2 1University of Minnesota Medical School, Minneapolis, MN, USA; 2Domestic Abuse Project, Minneapolis, MN, USA
OBJECTIVES/SPECIFIC AIMS: This study tests the feasibility of a 12-week yoga intervention designed to improve the mental health and coping skills of women receiving group therapy for intimate partner violence (IPV). Specific components tested included recruitment, safety, acceptability, and participant reaction. METHODS/STUDY POPULATION: A modified Hatha yoga routine was offered weekly for approximately 40 minutes at the end of the group session. A control group received typical group therapy. Questionnaires were administered at baseline and weekly throughout the course of the two groups. The questionnaires contained validated scales to measure the participants' mental health (depression, anxiety, PTSD, distress, perceived stress, hopelessness, and emotion regulation), exposure to IPV, social support, coping self-efficacy, self-care, and reaction to study participation. RESULTS/ANTICIPATED RESULTS: The study successfully recruited 85% (17/20) of those screened eligible (9 control, 8 intervention). Loss to follow up was 30% (5/17). No one reported emotional or physical harm. All intervention participants reported that the yoga was an appropriate level of difficulty, and that it was a meaningful contribution to group therapy. Both groups felt that their study-related rights were respected. 17% (2/12) thought the study questionnaire raised intense emotional issues for them; 92% (11/12) would participate again if they knew in advance what the study would be like. All participants thought the study was personally and socially beneficial (12/12). DISCUSSION/SIGNIFICANCE OF IMPACT: This study is a first step toward the development of a novel community-based intervention to mitigate the health effects of exposure to violence [Grant support includes 1UL1RR033183-01, 8UL1TR000114-02, and funds from the Program in Health Disparities Research].
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A LONGITUDINAL ANALYSIS OF QUALITY OF LIFE IN LONG-TERM CARE: IDENTIFYING POINTS FOR INTERVENTION
Shippee T1, Hong H1, Henning-Smith C1, Kane R1, Lewis T2 1University of MN, Minneapolis, MN, USA; 2Department of Human Services, Minneapolis, MN, USA
OBJECTIVES/SPECIFIC AIMS: This study examines resident-related and facility-level predictors of quality of life (QOL) in long-term care facilities in Minnesota from 2005 to 2010. The findings are modeled separately for low-, middle-, and high-performing facilities. METHODS/STUDY POPULATION: Data include self-reported QOL surveys from all NH facilities in Minnesota from 2005–2010 (N = 376); residents' clinical and payer data from the Minimum Data Set; and facility characteristics from the Department of Human Services. A total of 65,088 resident-level data are included. Analyses include factor analyses to identify QOL domains and two-level mixed effects linear models with random intercepts, modeling QOL domains over 5 years. RESULTS/ANTICIPATED RESULTS: Eight distinct domains of QOL were identified. These include: comfort, environment, autonomy, activity, dignity, meal enjoyment, relationships, and total score. These domains remained stable from 2005–2010, indicating reliability of the QOL measure. Findings show improvement in quality of care (QOC) but little change in QOL over time. Only “activity” domain showed consistent improvement over time. The scores for “autonomy”, “dignity”, and “relationships” domains tended to decrease from 2005–2010. Facility QOC score, size, ownership, and resident acuity were consistently significant predictors of QOL. Also, resident clinical conditions had a negative effect on QOL. DISCUSSION/SIGNIFICANCE OF IMPACT: Results indicate considerable variability in nursing home performance across different QOL domains, which needs to be taken into account when designing interventions. Results may be used to inform interventions to improve NH residents' satisfaction with QOL and to guide policy improvement efforts. Interventions must move beyond health-related QOC to more holistic programs designed to improve QOL.
CHARACTERIZING THE SELF-CARE “WORK” STRUCTURES AND PROCESSES OF CHRONIC HEART FAILURE PATIENTS
Holden R1 1Vanderbilt University School of Medicine, Nashville, TN, USA
OBJECTIVES/SPECIFIC AIMS: A tenet of the “human factors” discipline is that interventions must be informed by an evidence-based understanding of the to-be-improved work. Our objective was to analyze the structures and processes shaping the self-care “work” of chronic heart failure (CHF) patients. METHODS/STUDY POPULATION: Standardized survey, outpatient clinic and in-home observation, and semi-structured interview data from 27 CHF patients were analyzed using an adapted human factors work system model. RESULTS/ANTICIPATED RESULTS: Work system structure was decomposed into 5 interacting elements: person, tasks, tools/technology, organization, and environment. Person factors were related to both patients and informal caregivers, including abilities, limitations, and needs such as motivation and self-care knowledge. Task factors included the difficulty of self-care and established daily routines. Tool factors included pill boxes, diaries, and Internet. Key organization factors included social support and resource availability. The physical environment yielded barriers (stairs) and facilitators (desk) for self-care. Self-care processes included core care activities such as medication taking and nutrition but were inseparable from activities such as travel or chores. The processes were shaped by dynamic combinations of structural factors that made it easier or harder to perform the “work” needed to manage disease/health. I will present findings using summary diagrams and quotes/photos. DISCUSSION/SIGNIFICANCE OF IMPACT: Human factors offers a systematic way to depict complex work from multiple sources of data. Our results were as complex as the work described; such richness is needed to inform interventions so they properly fit or transform current work structures and processes. I will discuss how our findings are now being used to develop an IT intervention to support patients' self-care work.
EFFECTS OF DECISION SUPPORT ON THE PRESCRIPTION OF STATINS FOR THE PRIMARY PREVENTION OF CARDIOVASCULAR DISEASE (CVD)
Jean-Jacques M1, Long T2, Kaleba EO2, Dellahousaye J2, Lee J1, Persell SD1, Baker DW1 1Northwestern University Feinberg School of Medicine, Chicago, IL, USA; 2Alliance of Chicago Community Health Services, Chicago, IL, USA
OBJECTIVES/SPECIFIC AIMS: CVD is a leading cause of disparities in life expectancy. Statins are effective for the primary prevention of CVD among high risk patients, but less than half of eligible patients use statins, and rates of statin use are particularly low among minorities, the poor, and the uninsured. We evaluated the effects of decision support tools to promote the use of statins by patients at high risk for CVD who receive care within a network of community health centers (CHCs). METHODS/STUDY POPULATION: Decision support tools that facilitated the calculation of a patient's Framingham Risk Score (FRS), the prescription of generic statins, and the provision of low health literacy appropriate patient educational materials were implemented at 5 CHCs. Patients were eligible for the decision support tools if they were men ≥35 years or women ≥ 45 years, had no known coronary heart disease (CHD) or CHD equivalent, and if they did not have a statin on their active medication list. We compared the rate of statin prescriptions for eligible patients in the 6 months prior to and 6 months after the decision support implementation. RESULTS/ANTICIPATED RESULTS: The rate of new statin prescriptions did not change with implementation of the decision support tools:19% in the pre and post periods. The rate of statin prescriptions also did not differ when stratified by cardiovascular disease risk: 13% vs 14% for low risk, 21% vs. 22% for intermediate risk, and 31% vs. 32% for high risk patients in the pre vs post implementation periods. DISCUSSION/SIGNIFICANCE OF IMPACT: Provider-directed, point-of-care decision support tools did not improve the prescription of statins for primary CVD prevention among high risk patients.
EMERGENCY DEPARTMENT CROWING AND SHORT-TERM OUTCOMES FOLLOWING DISCHARGE
Gabayan GZ1, Derose SF2, Sun BC3 1UCLA, Los Angeles, CA, USA; 2Kaiser Permanente Southern California, Pasadena, CA, USA; 3Oregon Health and Science University, Portland, OR, USA
OBJECTIVES/SPECIFIC AIMS: Although Medicare will soon begin reporting Emergency Department (ED) crowding metrics, the impact of ED crowding is unclear. We hypothesized that a set of system and visit level measures of ED crowding are associated with bounce-back admission within 7 days of ED discharge. METHODS/STUDY POPULATION: We conducted a retrospective cohort study using data from a regional integrated health system, Kaiser Permanente Southern California for years 2008–2010 of members age 18 and over. We assessed system and visit level measures of ED crowding. System level metrics included exposures to ED occupancy, boarding time, and external length-of-stay (LOS). Visit metrics included waiting and evaluation time, as well as total LOS experienced by an index visitor. Covariates included demographic characteristics, comorbidities, Emergency Severity Index level, vital signs, ED discharge diagnosis, time variables, and ED site. For each crowding measure we fit linear and non-linear multivariable logistic regression models using quadratic cubic terms. RESULTS/ANTICIPATED RESULTS: The study cohort contained a total of 625,096 ED visits among 625,096 patients. There were 16,957 (2.7%) patients with a 7-day bounce-back admission. Compared to a median evaluation time of 2.2 hrs, an evaluation time of 10.8 hrs was associated with a relative risk of 3.9 (95% CI 3.7–4.1). Compared with a median ED LOS of 2.8 hrs, ED LOS of 11.6 hrs, was associated with a relative risk of 3.5 (95% CI 3.3–3.7). None of the other ED measures were associated with the outcome. DISCUSSION/SIGNIFICANCE OF IMPACT: Evaluation time and ED LOS are associated with increased 7 day bounce-back admissions. Our findings suggest that the Medicare measure of ED LOS in discharged patients is confounded by illness severity and is an unreliable measure of ED quality.
ESTABLISHING ATTENDING PHYSICIAN SAFETY AND QUALITY METRICS: SELECTION AND PILOTING OF NATIONAL PERFORMANCE MEASURES FOR INDIVIDUAL HOSPITALISTS
Michtalik HJ1, Pronovost PJ1, Miller RG1, Marsteller JA1, Brotman DJ1, Ford DE1 1Johns Hopkins University, Baltimore, MD, USA
OBJECTIVES/SPECIFIC AIMS: To determine criteria for selecting individual hospitalist metrics and to subsequently apply these criteria to currently collected and reported safety and quality measures to generate attending-physician-level metrics. METHODS/STUDY POPULATION: First, we used a series of interviews to perform a stakeholder analysis with 2 hospitalists, 3 hospital administrators, and 2 quality improvement experts to determine qualitative criteria for selecting appropriate attending-physician-level metrics. Next, we applied these criteria to publicly reported Maryland Potentially Preventable Complications (PPCs) and Joint Commission Core measures. Finally, we examined both the distribution and variability in these measures for regional hospitalist programs and Maryland hospitals. RESULTS/ANTICIPATED RESULTS: Stakeholders reached consensus that measures should be actionable, attributable, and accountable. Actionable was defined as a measure which could be actively intervened upon during a single hospital admission or a preventable adverse event. Attributable was defined as measures associated with, or under the supervision of, an attending physician. Accountability was defined as being within the responsibility of an attending physician. 34 of the 64 PPCs and 19 of the 50 Core measures reviewed met all three criteria. Regional review of PPCs over time showed variation both within and between hospitalist programs; Core measures at the statewide level showed a near normal distribution, indicating good discrimination amongst hospitals. DISCUSSION/SIGNIFICANCE OF IMPACT: Our study identified criteria to assign safety and quality measures to hospitalists. Regional and statewide analysis of these measures suggests adequate variation to compare individual hospitalists and programs both within and between institutions.
HOSPITAL READMISSIONS OF PEDIATRIC PATIENTS IN SOUTH CAROLINA: AN INITIAL ALL-HOSPITAL ANALYSIS
Scheurer M1, Robertson D1, Rees C1 1Medical University of South Carolina, Charleston, SC, USA
OBJECTIVES/SPECIFIC AIMS: 1) To determine statewide readmission rates for pediatric patients in South Carolina 2) To determine patient characteristics associated with pediatric readmission 3) To determine facility characteristics associated with pediatric readmission METHODS/STUDY POPULATION: State level data was obtained from the South Carolina Office of Research and Statistics for state fiscal years 2010 and 2011. The database is inclusive of all pediatric admissions to all 61 hospitals in South Carolina. Inclusions and Exclusions Index admissions of patients 18 years or less in state fiscal year 2010 were included. Normal neonatal admission and admissions for scheduled chemotherapy were identified by APR-DRG coding and excluded from analysis, Index admissions form 2010 were followed for additional hospital admissions in South Carolina through state fiscal year 2012. RESULTS/ANTICIPATED RESULTS: In South Carolina, there is significant geographic variability in readmission rates for children. Additionally, there is significant discrepancy in one year pediatric readmission rates between dedicated Children's and non-Children's Hospitals. Global one year readmission rates are significantly higher than readmission rates back to the same discharging hospital of the index admission. DISCUSSION/SIGNIFICANCE OF IMPACT: These analyses will be shared with the South Carolina Children's Hospital Collaborative with the intent of formulating actionable local and statewide QI projects to positively impact readmission rates.
LGBT APPALACHIANS AND TOBACCO USE: THE ROLE OF SOCIAL CONNECTEDNESS
Bennett K1 1University of Kentucky, Lexington, KY, USA
OBJECTIVES/SPECIFIC AIMS: 1. Understand themes related to lesbian, gay, bisexual, and transgender (LGBT) identity and how social contacts influence smoking and tobacco use behavior in an Appalachian population. 2. Identify modifiable barriers to tobacco prevention and cessation amongst rural LGBT persons. METHODS/STUDY POPULATION: Individual interviews with qualitative analysis and an online survey with descriptive and comparative quantitative analysis. Inclusion criteria are self-identified LGBT adults, both smokers and nonsmokers, who live in an Appalachian Kentucky county. RESULTS/ANTICIPATED RESULTS: Themes relating identity as an LGBT Appalachian to smoking and other health behaviors. The initial interviews will inform a quantitative, widely distributed survey targeted toward the same demographic for which data on tobacco use, health care access, and social interactions that influence health will be assessed. DISCUSSION/SIGNIFICANCE OF IMPACT: Research identifies health disparities that negatively impact lesbian, gay, bisexual, and transgender (LGBT) populations, including substance use. Additionally, LGBT people often face barriers to accessing care due to lack of health insurance, discrimination, and poor cultural competence of providers. LGBT persons residing in rural regions are likely to be at even higher risk. Elucidating the roles of peer pressure, social isolation, and group belonging is key to designing interventions that target the roots of risk-taking behavior. None of these have been explored in a rural LGBT population with regard to tobacco use. This study provides important information about a population for which health-specific information is almost non-existent. It prepares for more extensive research that will lead to targeted smoking cessation and prevention in this unique population.
LIVER TOXICITY MONITORING AND ITS IMPACT ON METHOTREXATE DISCONTINUATION IN A NATIONAL COHORT OF VETERANS
Schmajuk G1,2, Miao Y2, Yazdany J1, Steinman M1,2 1UCSF, San Francisco, CA, USA; 2San Francisco VA Medical Center, San Francisco, CA, USA
OBJECTIVES/SPECIFIC AIMS: Although the 2008 ACR guidelines advocate liver toxicity monitoring for patients receiving stable doses of oral methotrexate (MTX) every 8–12 weeks with an AST or ALT test, these guidelines are based on limited data from historical cohorts. METHODS/STUDY POPULATION: Using national data from the Veterans Health Administration (VHA), we assessed patterns, predictors, and consequences of liver function testing in a real-world cohort of patients taking oral MTX. Incident MTX users over 65 yo were linked to medical, pharmacy, and laboratory databases from 2007–2008. Inclusion criteria included ≥ 28-day supply of MTX. Exclusion criteria included evidence of having obtained care outside of the VHA (i.e. Medicare claims). Discontinuation was defined as lack of MTX dispensed for ≥ 90 days after the anticipated refill date. RESULTS/ANTICIPATED RESULTS: 717 MTX users were included, most were men (97%), mean age was 71 years (SD 6), mean MTX dose was 12.3 mg per week (SD 4.3). Only 28% of patients received monitoring consistent with ACR guidelines; most patients received monitoring < 3 times per year. Differences in adherence were related to health care utilization but not to other risk factors for MTX liver toxicity. Higher adherence to monitoring was associated with shorter time to the detection of an abnormal LFT (p < 0.001). More than 30% of patients with mild LFT elevations discontinued MTX, and many discontinuers went on to be treated with no DMARD (glucocorticoids or no treatment). DISCUSSION/SIGNIFICANCE OF IMPACT: Frequent monitoring of MTX users and the detection of potentially insignificant LFT elevations may have unintended consequences. A rational approach to monitoring targeted to patients at high risk of clinically meaningful liver toxicity should be developed.
PHYSICIAN SUPPLY AND HEALTH OUTCOMES: DO HAWAII COUNTIES WITH LOWER PHYSICIAN SUPPLY HAVE HIGHER READMISSION RATES?
Nagasako E1, Chen L1 1Washington University School of Medicine, St. Louis, MO, USA
OBJECTIVES/SPECIFIC AIMS: Hospital readmission is a health outcome relevant to patients, health care providers, health care purchasers, and policy makers alike; however the effect of different levels of physician supply on hospital readmission rates is unclear. This study compares readmission rates in Hawaii counties with varying levels of physician supply using administrative inpatient discharge data. METHODS/STUDY POPULATION: Population Studied: Adult Hawaii state residents with an inpatient discharge in 2007. This study uses data from the Healthcare Cost and Utilization Project (HCUP) State Inpatient Database (SID) for Hawaii to analyze relationships between physician supply and 30-day all-cause readmission rates. The Washington University School of Medicine Human Research Protection Office reviewed this study and determined that it did not contain activities subject to Institutional Review Board oversight. RESULTS/ANTICIPATED RESULTS: Counties in Hawaii with lower physician supply had lower, not higher, 30-day all-cause readmission rates compared to the county with the highest physician supply (City and County of Honolulu). Work is in progress to determine whether this difference is persistent even when factors such as age and number of comorbidities are taken into account. DISCUSSION/SIGNIFICANCE OF IMPACT: Lower readmission rates in counties in Hawaii with lower physician supply may reflect differences between patient populations, beneficial processes such as increased social support in these communities or may reflect unmeasured financial or nonfinancial barriers to accessing hospital care. Clarifying these mechanisms is important for reducing preventable readmissions. In addition to further quantiative research, qualitative methods which obtain patient and caregiver input such as key informant interviews and focus groups may be helpful in clarifying these processes.
RACIAL DISPARITIES IN MENTAL AND PHYSICAL HEALTH- RELATED QUALITY OF LIFE AMONG MEDICARE MANAGED CARE CANCER PATIENTS
Pandya C1, Li Y1, Fung C2, Mohile S2 1UofR, Rochester, NY, USA; 2Wilmot Cancer Center, Rochester, NY, USA
OBJECTIVES/SPECIFIC AIMS: To examine racial differences in health-related quality of life (HRQOL) outcomes and assess the association of socio-demographic, cancer-specific and general health characteristics with these outcomes among Black and White Medicare managed care patients diagnosed with cancer. METHODS/STUDY POPULATION: We used the Surveillance, Epidemiology, and End Results-Medicare Health Outcomes Survey (SEER-MHOS) linkage population-based database. The MHOS assesses measures of HRQOL through SF-36, which has physical component summary (PCS) score and mental component summary (MCS) score. Racial difference in median HRQOL scores was tested using Wilcoxon rank-sum test. Multivariable linear regression analysis was done to find the association between race and HRQOL after controlling for socio-demographic, cancer-specific and general health variables. 13,091 (9.1% Black vs. 90.9% White) patients diagnosed with breast, colorectal or prostate cancer are used. RESULTS/ANTICIPATED RESULTS: The median PCS and MCS scores for all cancer patients were 39 and 54 respectively. Black cancer patients had lower median PCS (36.7 vs 38.9 p < 0.00) and MCS (48.7 vs 51.3 p < 0.00) scores compared to White patients. In final regression model, PCS was lower (least-square mean: 38 vs. 40; p = 0.003) for Black compared to White patients. Interaction term between race and non-cancer comorbidities categories was significant (F-ratio = 3.09, p = 0.04). Lower PCS is associated with being female, older age, low income, low education, advanced cancer stage, recent cancer diagnosis, comorbidities, and having ADL deficits (all p < 0.05). DISCUSSION/SIGNIFICANCE OF IMPACT: Overall HRQOL for older Medicare cancer patients is very low in this study population. PCS scores are lower for Black compared to White patients. The racial disparities in HRQOL are likely to be moderated by number of non-cancer comorbidities.
SUPPORT SERVICE USE AND INTEREST IN SUPPORT SERVICES AMONG LUNG CANCER PATIENTS
Mosher C1, Winger JG1 1Indiana University-Purdue University Indianapolis, Indianapolis, IN, USA
OBJECTIVES/SPECIFIC AIMS: Lung cancer patients have high rates of clinically elevated distress and low rates of mental health service use. Little research, however, has examined correlates of mental health service use and interest in services in this population. Thus, we examined demographic and medical correlates of mental health service use and interest in services among lung cancer patients. METHODS/STUDY POPULATION: Lung cancer patients (N = 165) were recruited from two medical centers at least 3 weeks after their first oncology clinic visit. A total of 165 patients (51% male, 80% Caucasian, mean age = 63 years) participated in this phone survey. Fifty-seven percent had late-stage lung cancer, and the average time since diagnosis was 18 months. Patients completed the Hospital Anxiety and Depression Scale (HADS) and reported current mental health service use and interest in speaking with a staff member about their feelings. RESULTS/ANTICIPATED RESULTS: Twenty-eight percent of patients (47/165) reported current mental health service use. Furthermore, 40% of patients (16/40) with clinically elevated anxiety or depressive symptoms used these services. Women were more likely than men to use mental health services (odds ratio [OR] = 2.6, 95% CI = 1.3 to 5.3), and younger age was related to higher rates of service use (OR = 1.0, 95% CI = 0.9 to 1.0). Greater anxiety also was related to a higher likelihood of service use (OR = 1.1, 95% CI = 1.0 to 1.2). Only 9% of patients (11/118) who were not receiving mental health services were interested in speaking with a staff member. This interest was associated with being an ethnic minority group member (OR = 0.2, 95% CI = 0.1 to 0.8) and showing greater depressive symptoms (OR = 1.2, 95% CI = 1.0 to 1.4). DISCUSSION/SIGNIFICANCE OF IMPACT: Although lung cancer patients underused mental health services, those with higher anxiety were more likely to receive them.
THE RELATIONSHIP BETWEEN ELECTRONIC HEALTH RECORD USE AND OPIOID PRESCRIBING IN U.S. PRIMARY CARE OFFICE VISITS
Harle CA1, Cook RL1, Kinsell HS1, Harman JS1 1University of Florida, Gainesville, FL, USA
OBJECTIVES/SPECIFIC AIMS: Policymakers tout electronic health records (EHRs) as tools to help health care providers make appropriate treatment decisions. For example, EHRs may help primary care providers with the difficult decision of when to prescribe opioids for patients with chronic noncancer pain. This study assessed whether primary care visits to practices with EHRs were more likely to result in opioid prescriptions compared to visits to non-EHR practices. METHODS/STUDY POPULATION: We conducted a cross-sectional analysis of primary care visits in the 2007–2010 National Ambulatory Medical Care Surveys (NAMCS). We included all visits to an internal medicine or family/general practice provider by patients eighteen years or older who did not have cancer (N = 33,090). We used logistic regression to compare the odds of an opioid prescription among visits occurring in EHR practices versus non-EHR practice visits. RESULTS/ANTICIPATED RESULTS: Patients visiting practices with EHRs had significantly higher odds of receiving an opioid prescription compared to patients visiting non-EHR practices (OR = 1.31, p < .0001). Moreover, among the subset of patients with a chronic and common pain condition, EHR use was positively associated with opioid prescribing (OR = 1.47, p = .038). DISCUSSION/SIGNIFICANCE OF IMPACT: Primary care providers with EHRs were more likely to prescribe opioids. On one hand, EHRs could help providers confidently prescribe opioids by supporting tasks such as monitoring previous prescriptions and identifying side effects. On the other hand, physicians who are time-constrained by their EHRs could resort to opioid medications inappropriately because they have less time to address non-opioid approaches to pain management. The findings call for an increased understanding of how providers use EHRs to assist with pain management.
USING SIMULATION TO ASSESS THE INFLUENCE OF RACE AND INSURER ON PERIVIABLE COUNSELING: A WORK IN PROGRESS
Tucker Edmonds B1, McKenzie F1, Barnato A2, Frankel R1 1Indiana University School of Medicine, Indianapolis, IN, USA; 2University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
OBJECTIVES/SPECIFIC AIMS: To assess the effect of varying maternal sociodemographic characteristics on the content and quality of periviable counseling METHODS/STUDY POPULATION: We randomly assign obstetricians and neonatologists to encounter 2 comparable clinical scenarios of simulated patients diagnosed with ruptured membranes at 23 weeks, systematically varying patient race (black/white), insurer (private/Medicaid), and the order/combination in which the cases are presented. Counseling interactions are observed, video-, and audio-recorded. We will assess communication content and quality using content analysis; then compare these across race, insurer, and specialty. To-date, we have completed case development with a multidisciplinary team; conducted extensive standardized patient training; and performed three pilot sessions to test the cases. Recruitment and code book development are now underway. We have currently recruited 12 of 16 obstetricians and 10 of 16 neonatologists and have conducted 19 total simulation sessions. RESULTS/ANTICIPATED RESULTS: We anticipate finding that less shared decision-making occurs among black and Medicaid-insured patients. Across specialty, we expect to see marked variation in the consistency of prognostic information delivered. DISCUSSION/SIGNIFICANCE OF IMPACT: Pregnant women making resuscitation decisions for periviable infants rely on physicians' counseling to inform those decisions. Counseling should be unbiased and patient-centered to align treatment plans with patients' goals and values. This pilot data will inform the subsequent development of counseling interventions and decision support to minimize disparities in informed decision-making and optimize shared decision-making in periviable care.
VISIT PATTERNS & CHILD OUTCOMES IN THE NURSE-FAMILY PARTNERSHIP PROGRAM
Holland ML1, Crean HF1, Olds DL2, Kitzman H1, Dozier AM3 1University of Rochester, Rochester, NY, USA; 2University of Colorado Denver, Denver, CO, USA; 3University of Rochester – Public Health Science, Rochester, NY, USA
OBJECTIVES/SPECIFIC AIMS: To identify home visit adherence patterns in an evidence-based intervention for low-income first-time mothers and examine if these patterns are associated with outcomes. METHODS/STUDY POPULATION: Data from a Nurse-Family Partnership trial conducted in Memphis, TN (1990–91). We examined visit patterns of these primiparous women on Medicaid (n = 212). Using latent class analysis, we tested associations between probability of belonging to each latent class and 12 year child outcomes: internalizing disorders (Youth Self-Report) & Peabody Individual Achievement Test scores for math and reading. RESULTS/ANTICIPATED RESULTS: Preliminary findings identified four adherence classes with good separation (entropy = .82). Members of Class 1 (23% of the participants) attended an average of 104% of the recommended visits; Class 2 (28%): 71% of visits; Class 3 (29%): 55% of visits; Class 4 (20%): 28% of visits. Children of mothers in Class 2 had the lowest likelihood of internalizing disorders (Class 1: Relative Risk (RR) = 3.7, p < .01; Class 3: RR = 2.4, p < .08; Class 4: RR = 2.7, p < .04). Children of low-psychological-resource mothers in Class 1 had higher math scores than those in Class 3 (standardized beta [sb] = .56, p < .06) and Class 4 (sb = .63, p < .09) and higher reading scores than those in Class 3 (sb = .63, p < .05). DISCUSSION/SIGNIFICANCE OF IMPACT: Distinct visit patterns were associated with different outcomes. Retention is an important issue for many interventions translated from research settings. Next, we will examine patterns in dissemination sites where attrition rates are up to 60%. Understanding visit patterns may help programs focus early retention efforts on mothers at risk of falling into Classes 3 and 4, whose children may have worse outcomes.
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A CUBIIC MODEL FOR CORRECTING GENE EXPRESSION LEVELS FOR PATIENT AGE IN CONTROL TREATMENT STUDIES
Fongang B1, Finnerty CC1,2, Li X1, Tompkins RG3, Herndon DN1,2, Kudlicki AS1,2 1University of Texas medical Branch, Galveston TX, TX, USA; 2Shriners Hospitals for Children, Galveston, TX, USA; 3Massachussets General Hospital, Boston, MA, USA
OBJECTIVES/SPECIFIC AIMS: An important component in clinical and translational studies is whole-transcriptome expression profiling of tissue samples. In rare diseases or injuries, as severe burns, it is impossible to maintain a homogeneous group of subjects, which may result in increased inherent variance due to factors as age or sex, which will cause artificially increased p-values for detection of differential expression between the control and treatment groups. However, a model of the age-dependence for every transcript can help overcoming the problem by comparing the expression profile in a treatment subject to the expected profile for a person of the same age, interpolated from the control data. METHODS/STUDY POPULATION: We analyzed 61 whole-genome profiles of white blood cells in healthy volunteers between the ages of 8 and 63, including 34 females and 27 males. The profiles were collected following an internally consistent protocol using the Affymetrix GeneChip (R) Human Genome U133 Plus 2.0 microarray platform. RESULTS/ANTICIPATED RESULTS: We observe that the cubic root of age is most suitable for linear modeling and interpolation, and we show that the Pearson correlation coefficient between cubic root of age and transcript concentration is significant for 9,848 out of the 54,675 probesets on the U133 Plus 2.0 array, (0.05 confidence after FDR correction). DISCUSSION/SIGNIFICANCE OF IMPACT: Applying our cubic correction reduces the control variance by up to 70%.
A MULTI-OMICS STRATEGY FOR IDENTIFYING NOVEL BIOMARKERS AND DISEASE MECHANISMS OF RECURRENT CLOSTRIDIUM DIFFICILE INFECTION
Dann SM1, Hollister E2, Oezguen N2, Urvil P2, Eaves-Pyles T1, Loeffelholz M1, Luna R2, Jiang Z3, DuPont H3, Versalovic J2, Savidge T2 1University of Texas Medical Branch, Galveston, TX, USA; 2Texas Children's Microbiome Center and Baylor College of Medicine, Houston, TX, USA; 3University of Texas School of Public Health, Houston, TX, USA
OBJECTIVES/SPECIFIC AIMS: The incidence of Clostridium difficile infection (CDI) has increased dramatically. Although vancomycin and metronidazole are preferred treatment options, neither is fully effective and development of disease recurrence is common and costs the U.S. healthcare system over $3.8 billion annually. Reduction in recurrence, therefore, is a priority clinical need. METHODS/STUDY POPULATION: To identify biomarkers associated with CDI pathogenesis, we profiled stool microbial composition and the metabolome from 55 patients with and without CDI and performed cross-omics analysis. RESULTS/ANTICIPATED RESULTS: Patients that develop recurrent disease have significant shifts in microbial populations and biochemical composition. We found decreased abundances of taxa belonging to the families Bacteroidaceae and Prevotellaceae, and increased abundances of taxa belonging to the families Peptostreptococcaceae and Clostridiaceae. Concurrent with changes in microbial composition, significantly elevated levels of the neurotransmitter γ-aminobutyric acid (GABA) were measured. Using agonists and antagonists of GABA, we demonstrate that GABAergic signals directly regulate C. difficile neurotoxin activity and cytokine secretion in experimental CDI. DISCUSSION/SIGNIFICANCE OF IMPACT: We demonstrate that integration of global metabolomics and metagenomics into network models provides framework for the discovery of new diagnosis and treatment in CDI. The significance of this multi-omics approach is our identification of a novel microbe-neuroimmune signaling mechanism in patients with recurrent disease.
A NOVEL STATISTICAL MODEL FOR FAMILY-BASED CASE-CONTROL STUDIES OF GENETIC IMPRINTING
Wang Z1 1Yale University, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Genetic imprinting arises from a gene when either its maternally- or paternally-derived copy is expressed while the other copy is silenced. In this study, we present a statistical model for testing the effect of genetic imprinting (or parent-of-origin) on a human disease in case-control studies with family structure. METHODS/STUDY POPULATION: For each subject sampled from a case and control population, we not only genotype its own single nucleotide polymorphisms (SNPs), but also collect its parents' genotypes. By tracing the transmission pattern of alleles from parental to offspring generation, the model allows the characterization of genetic imprinting effects based on chi-squared tests of a contingency table. The model is expanded to test the interactions between imprinting effects and additive, dominant, and epistatic effects in a complex web of genetic interactions. RESULTS/ANTICIPATED RESULTS: We investigated the statistical properties of the model and its practical usefulness is validated by a real data analysis. DISCUSSION/SIGNIFICANCE OF IMPACT: Genetic imprinting has been recognized to play an important role in the formation and pathogenesis of human diseases. The model proposed here provides a powerful statistical and computational tool for detecting genes that determine complex diseases in an imprinting way. It will help to elucidate a detailed picture of the genetic architecture for developmental disorders.
A WIRELESS HEAD-MOTION MONITORING AND TELEREHABILITATION SYSTEM FOR IMPROVING OUTCOMES IN VESTIBULAR REHABILITATION
Bhatti P1,2, Datta Roy S1, Heusel-Gillig L2, Tusa R2, Herdman S2 1Sch. of Elec. & Comp. Engin., Georgia Tech., Atlanta, GA, USA; 2Dept. of Rehab. Med., Emory Univ., Atlanta, GA, USA
OBJECTIVES/SPECIFIC AIMS: This work reports a wireless head-motion monitoring system (HAMMS) to record and transmit patient head movements while completing exercises for vestibular rehabilitation. The aim is to assess the quality of exercise (amplitude, frequency, velocity) during in-clinic and in-home exercise sessions. We hypothesize that patients underperform exercise routines and therefore experience poor outcomes. METHODS/STUDY POPULATION: We focus on critical gaze stabilization exercises for patients with unilateral (UVH) or bilateral vestibular (BVH) hypofunction: adaptation and substitution exercises. The HAMMS captures 3D head rotation and provides both a qualitative and quantitative means to assess exercise. The HAMMS can also wirelessly transmit data to a patient's tablet or smartphone during in-home exercises. These data can be conveniently uploaded to a database for a clinician's review. The clinical can provide nearly realtime feedback via the tablet/smartphone thereby remotely adjusting the exercise plan. RESULTS/ANTICIPATED RESULTS: Movements were completed in the yaw and pitch planes for one minute each while fixating on a target at 8-ft. To observe group differences, a power spectral density was calculated for 5 UVH and 5 normals. In the yaw plane UVH patients demonstrated an avg. frequency of 0.94 Hz (sd. 0.23) and 1.52 Hz (sd. 0.25) for normals. In the pitch plane UVH patients demonstrated an avg. frequency of 1.01 Hz (sd. 0.46), and 1.54 Hz (sd. 0.43) for normals. DISCUSSION/SIGNIFICANCE OF IMPACT: We have developed a system that can be used to monitor head motion for patients performing vestibular rehabilitation exercises. The HAMMS as an intervention may improve exercise quality, adherence, and the ability to increase clinician interaction through a tele-rehabilitation interface.
AN INTERACTIVE VOICE RESPONSE DIARY FOR STIFFNESS IN NON-DYSTROPHIC MYOTONIA
Barohn R3,9, Statland J1, Bundy B2, Wang Y3, Raja Rayan D4, Trivedi J5, Sansone V6, Salajegheh M7, Venance S8, Ciafaloni E1, Matthews E4, Meola G6, Herbelin L3, Griggs R1, Hanna M4 1University of Rochester Medical Center, Rochester, NY, USA; 2University of South Florida, Tampa, FL, USA; 3University of Kansas Medical Center, Kansas City, KS, USA; 4UCL Institiute of Neurology, London, United Kingdom; 5University of Texas Southwestern, Dallas, TX, USA; 6University of Milan, Milan, Italy; 7Brigham and Women's Hospital, Boston, MA, USA; 8London Health Sciences Centre, London, ON, Canada; 9for the Consortium for Clinical Investigation of Neurologic Channelopathies, Rochester, NY, USA
OBJECTIVES/SPECIFIC AIMS: To determine the minimal clinically important change (MCIC) of an automated telephone-based interactive voice response diary (IVR) of patient-reported stiffness for non-dystrophic myotonia (NDM) using data from a recently completed randomized controlled trial of mexiletine in NDM. METHODS/STUDY POPULATION: Stiffness was rated by each participant on a 1–9 scale, 1 being minimal, and 9 the worst ever experienced. Responsiveness was defined as the within subject change in an outcome divided by the standard deviation. The SF36 preference-based health utility index (question 2) was used as an anchor to determine the MCIC of stiffness on the IVR. An increase in 1 corresponds with rating health 'somewhat better now than one year ago.' RESULTS/ANTICIPATED RESULTS: Stiffness on the IVR was the most responsive outcome in the clinical trial, with an effect size of 1.36. The MCIC of stiffness on the IVR was 2.51 (95% confidence interval [CI] 1.22, 3.83). A decrease of 1 on graded myotonia by electromyography corresponded with a decrease of 2.12 on the IVR stiffness scale (95% CI 1.24, 3.00). For every 1 second decrease in eye closure myotonia on bedside exam stiffness on the IVR decreased by 0.34 (95% CI 0.03, 0.65). DISCUSSION/SIGNIFICANCE OF IMPACT: Stiffness on the IVR was responsive in NDM, correlated with other measures of myotonia, and 1.22 to 3.83 were the lower and upper limits of MCIC.
COMPARISON OF SCREENING METHODS FOR CARBAPENEM RESISTANT ENTEROBACTERIACEAE
Ryder A1, Alhazmi A2, Somily A2, Peaper D1,3, Murray T1 1Yale University School of Medicine, New Haven, CT, USA; 2King Saud University and King Khalid University Hospital, Riyadh, Saudi Arabia; 3VA Connecticut Healthcare System, West Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Severe infections caused by multi-drug resistant gram negative rods (MDR-GNR) including carbapenem resistant Enterobacteriaceae (CRE) are a source of significant healthcare costs and mortality. CRE infections are becoming increasingly prevalent, prompting many institutions with at-risk patients to consider implementing routine screening of certain patient populations. A laboratory-based surveillance program to identify hospitalized patients colonized by CRE represents an important component of an overall strategy to minimize the spread of resistant bacteria. METHODS/STUDY POPULATION: Several CRE screening methods have been described, and their performance has been evaluated primarily in the context of CRE outbreaks, or in populations of high CRE colonization prevalence; however, there is no consensus as to which method is most appropriate for identifying colonized patients in low-prevalence settings. The purpose of this study was to directly compare the performance characteristics of available methods of detecting gastrointestinal colonization by CRE in a non-outbreak setting. RESULTS/ANTICIPATED RESULTS: We found that chromogenic agar plates formulated to detect bacteria expressing extended spectrum β-lactamases were the least time consuming and most sensitive procedure for identifying colonization by CRE in this hospitalized patient population. The recommended Centers for Disease Control screening method demonstrated comparatively poor sensitivity and specificity. DISCUSSION/SIGNIFICANCE OF IMPACT: Understanding the performance of available screening methods, as well as the limitations associated with each of these methods in settings of both high and low CRE prevalence, is essential when considering implementing a large-scale CRE surveillance program.
COPDFLIX: A SOCIAL MEDIA RESOURCE CENTER FOR PATIENT-CENTERED SELF-MANAGEMENT TRAINING
Stellefson M1, Chaney B1, Walsh-Childers K1, Sriram P1, Chaney D1 1University of Florida, Gainesville, FL, USA
OBJECTIVES/SPECIFIC AIMS: The objective of this ongoing research is to conceptualize the design for a patient-centered social media resource center that can be used by individuals with COPD. METHODS/STUDY POPULATION: A multidisciplinary group of researchers from multiple fields (Health and Human Performance, Journalism and Communications, and Medicine) have collaborated to conduct key informant interviews with healthcare experts and individuals with COPD to provide insight for social media development. The next step in our research will be to test the COPDFLIX Social Media Resource Center (CSMRC) for acceptability, appropriateness, and usability among a sample of 15–25 individuals with COPD. Eligible participants will be asked to participate in 30- to 45-minute usability testing sessions that will be recorded using audio-video and Camtasia screen capture technology. Participants will also be asked to provide responses to a 16-item acceptability and usability scale, which will evaluate users' perceptions of all social media features included on the website. RESULTS/ANTICIPATED RESULTS: Semi-structured interviews with expert consultants and individuals with COPD have revealed numerous insights for strengthening the utility of the CSMRC. We anticipate the forthcoming usability testing sessions will provide the research team with rich data on user performance (i.e., ease of use, efficiency, errors) and satisfaction. This data will subsequently be used to make further design refinements to CSMRC. DISCUSSION/SIGNIFICANCE OF IMPACT: Our current research will likely lead to novel approaches for empowering, engaging, and educating older adults with COPD. This type of formative health communication research is necessary before future efficacy trials investigate the comparative effectiveness of online pulmonary rehabilitation training versus standard of care programs.
DETECTION OF ISLET BETA CELL-SPECIFIC DNA USING MULTIPLEX PCR ANALYSIS IN SERUM SAMPLES TO PREDICT DIABETES PROGRESSIOND
Tersey S1, Fisher M1, Mather K1, Mirmira R1 1Indiana University School of Medicine, Indianapolis, IN, USA
OBJECTIVES/SPECIFIC AIMS: Non-invasive detection of early islet β cell death in type 1 and type 2 diabetes might identify individuals in whom interventions would preserve β cell mass and prevent hyperglycemia. Recent studies have shown that dying β cells in mice are correlated with an increase in the appearance of unmethylated preproinsulin (PPI) DNA in the serum. METHODS/STUDY POPULATION: We developed a dual fluorescent probe-based multiplex PCR assay (TaqMan) to detect changes in differentially methylated PPI DNA in cell lines, tissues, and serum. The “methylation index” was defined as a parameter that reflects the relative predominance of methylated versus unmethylated PPI DNA. The methylation index was measured in the serum of several diabetic mouse models, including high dose and multiple low dose streptozotocin-induced diabetes, the NOD mouse model of type 1 diabetes, and the db/db mouse model of type 2 diabetes. RESULTS/ANTICIPATED RESULTS: Key assay features included zero to low background signals, assay output that is linear, and simultaneous detection of methylated and unmethylated PPI DNA species in a single reaction. Our data show that the methylation index decreases significantly in the days or weeks prior to the development of hyperglycemia, and increases to baseline levels or higher following the development of frank diabetes, when no β cells are detectable in pancreatic sections. We anticipate that the assay with slight modifications will work to detect progression to human disease. DISCUSSION/SIGNIFICANCE OF IMPACT: In conclusion, the methylation index in serum is a sensitive indicator of dying β cells, and may be useful to track diabetes progression and response to therapeutic intervention.
DEVELOPMENT OF PERSONALIZED MICROBIOME CULTURE COLLECTIONS
Goodman AL1 1Yale University, West Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Advances in DNA sequencing technology have shed new light on the range of variation that exists in the human gut microbiome. However, distinguishing causation from correlation remains a central challenge in the field. METHODS/STUDY POPULATION: To address this, we have developed personalized, arrayed gut microbiome culture collections from individual human donors. RESULTS/ANTICIPATED RESULTS: These collections include phylum-, class- and order-level representatives for 99% of the donor gut microbial community, and genus-level representatives for >70% of this community. Shotgun metagenomic sequencing of these collections reveals that 90–95% of annotated functions (e.g., KEGG orthology and pathway annotations, Enzyme Commission assignments) identified in a complete, uncultured microbiome are also observed in a personalized culture collection derived from that microbial community. DISCUSSION/SIGNIFICANCE OF IMPACT: Because each isolate in the collection is individually archived, this approach allows re-assembly of a gut microbiome in a manner that key microbes (or groups of microbes) can be omitted. This should allow direct testing of connections between the presence or abundance of key microbial phylotypes and associated host phenotypes.
DIMENSIONAL CLASSIFICATION OF SCHIZOPHRENIA (D.C.).THE NEUROCOGNITIVE ENDOPHENOTYPES (N.E.)
Estani P1 1Professor Dr. Leon S. Morra Medical Hospital., Córdoba, Argentina
OBJECTIVES/SPECIFIC AIMS: The actual classifications of mental disorders by multiple categories, such as the DSM IV TR, (APA, 1994), have stopped being effective, specially in the field of the new psychiatric genetics, neurobiological investigations of schizophrenia and mental disorders in general, because they do not fit in our actual neurobiological data in field. At the moment we are looking for simple, dimensional classifications (D.C.) cradles in the concept of endophenotype, as this concept was defined by Dr. Gottesman in its application to psychiatric research. (Gottesman, 2003). METHODS/STUDY POPULATION: A total of 1200 participants (patients with a diagnosis of schizophrenia and normal controls) were the subjects. Neuropsychological tests were applied. We operationalized two neurocognitive endophenotypes (N.E.) : sensorial gating and cognitive performance RESULTS/ANTICIPATED RESULTS: The dimensional classification permits the inclusion of the mechanism of neurocognitive endophenotypes. DISCUSSION/SIGNIFICANCE OF IMPACT: The dimensional classifications of mental disorders allow full inclusion of the concept of endophenotypes,defining the genotypes to benefit the understanding and treatment of schizophrenia. The full explanation of this categorization will be presented along with the results of this pilot sample.
IDENTIFYING FALLS IN AN EHR-BASED COHORT
Womack JA1, Garla V3, Brandt CA2,3 1Yale School of Nursing, New Haven, CT, USA; 2Veterans Administration Connecticut Health Services, West Haven, CT, USA; 3Yale Center for Medical Informatics, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: External Cause of Injury (E)codes identify injuries such as falls in administrative datasets. However, falls are undercoded. We will explore whether Information about falls is documented in progress notes or radiology reports. METHODS/STUDY POPULATION: We identified individuals with falls Ecodes in 2009 as well as radiology reports and progress notes for these individuals that occurred within 1 month of the E-code date. We also identified individuals at risk for a fall but without Ecodes in 2009. We identified individuals with ICD9 codes for conditions that might place them at risk for a fall. We then selected 20% of the radiology reports and 20% of the progress notes from these individuals. All notes were reviewed and classified as fall/not fall. A randomly selected subset was reviewed by another clinician to confirm categorization. These notes were the reference standard. Notes were pre-processed to identify individual words, parts of speech, negation, etc. A bag of words model was used to identify notes that addressed falls vs those that did not. This model represents texts as an unordered collection of words. The frequency of each word is used as a feature for training a classifier. RESULTS/ANTICIPATED RESULTS: Radiology reports for 276 individuals were reviewed: 1200 notes from those with a falls Ecode, and 7000 notes for those without. Progress notes for 81 individuals were reviewed: 2892 notes for those with an E-code and 1015 notes for those without. For radiology reports, sensitivity of the algorithm was 0.86; positive predictive value 0.91; F score 0.88; Kappa 0.87. For progress notes, sensitivity of the algorithm was 0.93; PPV 0.98, F score 0.95; and Kappa 0.86. DISCUSSION/SIGNIFICANCE OF IMPACT: Radiology reports and progress notes may provide information about falls in the VHA electronic medical record.
INCREASING THE EVIDENCE BASE FOR USING COMMON FACTORS IN PEDIATRIC PRIMARY CARE
Polk S1, Longway S2, Fothergill K2, Wissow L2 1Johns Hopkins School of Medicine, Baltimore, MD, USA; 2Johns Hopkins School of Public Health, Baltimore, MD, USA
OBJECTIVES/SPECIFIC AIMS: To better understand how to engage pediatric patients during primary care by examining; 1) children's perspectives on physician communication; 2) children's perspectives on physician/parent communication; and 3) if and how these perspectives vary when there is language discordance between provider and parent. METHODS/STUDY POPULATION: This observational pilot study included 32 8–11 year-old patients at an urban, general pediatrics practice serving primarily publicly-insured children. Well-child visits were video-taped and used in a stimulated-recall interview with the patient to solicit opinions regarding patient-provider engagement. Interview transcripts were analyzed for major themes related to common factors. RESULTS/ANTICIPATED RESULTS: 32 children were enrolled, 16 Spanish-speaking Latinos, 8 English-speaking Whites and 8 English-speaking Blacks. Participants reported enjoying being asked questions by their clinician, particularly regarding school. Participants expressed interest in advice about health. Fear of vaccines and negative anticipation of the physical exam, especially among males, distracted participants during their visit. Opinions varied regarding how the clinician should divide their attention between the patient and their parent. Children in Latino families reported that their parents could understand the clinician, but described triangulation i.e. the provider and patient understood each other in English only and the provider and parent understood each other in Spanish only. DISCUSSION/SIGNIFICANCE OF IMPACT: Participants were interested in discussing their health with and learning about health from their clinician. An improved understanding of how to engage school-aged pediatric patients is a step towards the expansion of mental health care in pediatric primary care practice.
INFLUENZA FORECASTING WITH GOOGLE FLU TRENDS
Dugas AF1, Jalalpour M1, Gel Y1,2, Levin S1, Torcaso F1, Igusa T1, Rothman R1 1Johns Hopkins University, Baltimore, MD, USA; 2University of Waterloo, Waterloo, ON, Canada
OBJECTIVES/SPECIFIC AIMS: We sought to develop a practical influenza forecast model, based on real-time, geographically focused, and easy to access data, to provide individual medical centers with advanced warning of the number of influenza cases, thus allowing sufficient time to implement an intervention. METHODS/STUDY POPULATION: Forecasting models designed to predict one week in advance were developed from weekly counts of confirmed influenza cases over seven seasons (2004 – 2011) divided into training and out-of-sample verification sets. Forecasting procedures using classical Box-Jenkins, generalized linear, and autoregressive methods were employed to develop the final model and assess the relative contribution of external variables such as, Google Flu Trends, meteorological data, and temporal information. Models were developed and evaluated through global forecast deviance. RESULTS/ANTICIPATED RESULTS: A generalized autoregressive Poisson (GARMA) forecast model integrating previous influenza cases with Google Flu Trends information provided the most accurate influenza case predictions. Google Flu Trend data was the only source of external information providing significant forecast improvements (p = 0.00002). DISCUSSION/SIGNIFICANCE OF IMPACT: Integer-valued autoregression of influenza cases provides a strong base forecast model, which is enhanced by the addition of Google Flu Trends confirming the predictive capabilities of search query based syndromic surveillance. This accessible and flexible forecast model can be used by individual medical centers to provide advanced warning of future influenza cases.
LESSONS LEARNED: RECRUITMENT OF OLDER AFRICAN AMERICAN MALES FOR DEPRESSION RESEARCH
Bryant K1, Newsome Wicks M2, Willis N1 1University of Arkansas for Medical Sciences, Little Rock, AR, USA; 2University of Tennessee Health Science Center, Memphis, TN, USA
OBJECTIVES/SPECIFIC AIMS: Higher levels of depression have been linked to psychosocial stressors and negative life events, and thus African American men in particular may have a greater propensity for depression than the majority population. The aim was to describe lessons learned from the recruitment efforts of older African American men in depression research. METHODS/STUDY POPULATION: Participation was sought by targeting community clinics that served African American men in a Mid-South metropolitan area. In addition, flyers were distributed and posted in community locations including churches and businesses, such as barbershops, that catered to predominately African American male clientele. Despite these recruitment strategies the study yielded no participants after nine months of recruitment efforts. RESULTS/ANTICIPATED RESULTS: The research team consulted with university community liaisons and a local community group to discuss the study and explore recruitment strategies. Meeting attendees acknowledged the need for and importance of the study and ascribed the lack of recruitment success as likely due to multiple barriers. Some of the meeting attendees' comments related to the stigma associated with depression. With regard to the protocol, meeting attendees took issue with the requirement for a formal diagnosis of depression because they believed that many eligible men had not been formally diagnosed with depression by a healthcare provider. They also provided valuable feedback on the recruitment flyer. The attendees recommended word of mouth as the best recruitment approach because of the stigma associated with depression and the lack of trust in the university. DISCUSSION/SIGNIFICANCE OF IMPACT: These recruitment strategies may the key to a successful mental health study among African American males.
MEASUREMENT BIAS IN PEDIATRIC BEHAVIORAL SCREENING TOOLS
Studts CR1 1University of Kentucky, Lexington, KY, USA
OBJECTIVES/SPECIFIC AIMS: Early identification of child disruptive behavior problems may help reduce the serious long-term consequences of this public health problem. Screening tools like the PSC-17 and the BPI may be valuable for prevention, but disparities in results among female, minority, and low SES children have been reported. Elimination of measurement bias could lead to improved behavioral screening, reducing disparities in identification rates among diverse groups of children. Previous psychometric studies have relied on traditional methods which cannot adequately ascertain bias. METHODS/STUDY POPULATION: Parents (N = 900) of 3–5 year old children (53% male, 50% white, 43% low SES) from 4 diverse primary care settings completed the PSC-17 and BPI. Item response theory (IRT) analyses tested items for differential item functioning (DIF) by child sex, race, and SES. Two approaches to DIF-detection were used. Both methods' results were compared to reveal items with high levels of bias. RESULTS/ANTICIPATED RESULTS: Of 18 items, 8 exhibited statistically significant DIF (p < .0027, corrected for multiple comparisons). Bias by child sex was found in 2 items (refuses to share, breaks/destroys things); by child race, in 4 (blames others, teases others, high strung, argues too much); and by child SES (low versus medium/high), in 3 (high strung, cheats/lies, stubborn/sullen/irritable). DIF by race and SES significantly impacted overall behavioral scores (p < .001). DISCUSSION/SIGNIFICANCE OF IMPACT: Equitable early identification in diverse populations of young children requires screening questions to be unbiased. The 8 identified PSC-17 and BPI items may be inappropriate for use in behavioral screening of young children. Follow-up studies should (a) identify the sources of DIF (e.g., differences in parent perceptions vs. actual disparities in child behavior between groups), and (b) develop DIF-free behavioral screening tools for early identification.
MOLECULAR MRI OF MULTIPLE SCLEROSIS USING 7 TESLA
Dula AN1, Smith SA1, Sriram SM1, Gore JC1 1Vanderbilt University, Nashville, TN, USA
OBJECTIVES/SPECIFIC AIMS: Current radiological markers remain insufficient at capturing pathology of multiple sclerosis (MS): noninvasive methods lack pathologic specificity. While imaging studies typically concentrate on focal lesions, we will quantitatively measure tissue properties reflecting the dynamic and heterogeneous tissue microenvironment in normal appearing white matter (NAWM). METHODS/STUDY POPULATION: CEST MRI was optimized for brain and spinal cord (SC) application to MS patients at 7T through tissue-specific simulation of the expected amide proton CEST effect. Protocols were further optimized empirically and applied to the brain and SC in 10 healthy and relapsing-remitting (RR) MS patients. White matter (WM) was segmented using fully automated, topology-preserving approach. Histograms of segmented APT metrics were compared between healthy and MS subjects. RESULTS/ANTICIPATED RESULTS: We have evaluated the entire central axis of the nervous system in the context of the molecular manifestation of MS. Data suggest that the median APT metrics of NAWM in the brain and cervical spine are higher in MS patients relative to healthy participants. Additionally, in ongoing studies, we anticipate close correlation of the APT measure, particularly in the spinal cord, with clinical measures of disability. DISCUSSION/SIGNIFICANCE OF IMPACT: Chemical Exchange Saturation Transfer (CEST) MRI is a molecular imaging method sensitive to metabolites, reflecting tissue composition based on distinct solute/water interactions. CEST MRI is sensitive to protein content (amide proton transfer) with potential to elucidate the largely undetected pathology of NAWM. Thus 7T CEST MRI reports on subtle molecular pathology of NAWM potentially clarifying clinical manifestations poorly explained by macroscopic lesions.
PARKINSON DISEASE BIOMARKERS ASSAY DEVELOPMENT AND PATIENT ENROLLMENT
Rosenthal L1, Jelinek C1, Salnikova Y1, Nichols P1, Dorsey R1, Bakker C1, Bassett S1, Dawson T1 1Johns Hopkins, Baltimore, MD, USA
OBJECTIVES/SPECIFIC AIMS: Post-translational modifications at six sites within alpha-synuclein have been identified in the post-mortem brain tissue of Parkinson disease (PD) patients. This investigation seeks to determine the value of these modifications as biomarkers in the cerebrospinal fluid (CSF). METHODS/STUDY POPULATION: First, we used CSF from standardized healthy control patients to create a selected reaction monitoring assay and quantify the relative abundance of these six posttranslational modifications in these controls. Second, we are enrolling healthy controls and individuals with PD in a prospective, observational study. Each participant undergoes a comprehensive history, physical and neurological examination, and biological specimens collection via venipuncture and spinal tab. RESULTS/ANTICIPATED RESULTS: We have successfully developed the assay to assess the value of these posttranslational modifications as biomarkers. Our multiplexed selected reaction monitoring assay consists of six scheduled targeting experiments and takes 32 minutes to complete. Sample preparation is performed using the Perfinity Workstation, an automated sample preparation platform. Using this assay, we confirmed our ability to detect these six posttranslational modifications in the CSF of our standardized healthy control patients. Eight participants are currently enrolled in our prospective, observational study and three additional individuals are scheduled for their first visit. DISCUSSION/SIGNIFICANCE OF IMPACT: Modifications to alpha-synuclein can be identified in the CSF using selected reaction monitoring mass spectrometry. More data are needed to determine if the modified peptides can differentiate between PD patients and controls.
PLASTICITY OF CORTICAL ACTIVITY AND CONNECTIVITY DURING BRAIN-COMPUTER INTERFACE LEARNING AND CLINICAL IMPLICATIONS
Wang W1, Degenhart AD1, Wang D2,1, Tyler-Kabara EC1, Collinger JL1,3, Boninger ML1,3 1University of Pittsburgh, Pittsburgh, PA, USA; 2Zhejiang University, Hangzhou, China; 3Human Engineering Research Laboratories, Department of Veterans Affairs, Pittsburgh, PA, USA
OBJECTIVES/SPECIFIC AIMS: Brain-computer interface (BCI) technology aims to enable individuals with limb paralysis to control assistive/robotic devices with only brain signals. Previous basic BCI studies done in animals suggested that cortical plasticity might play a critical role in learning to operate BCI devices. METHODS/STUDY POPULATION: We are uniquely positioned to study such cortical plasticity in a clinically relevant context. Previously, we have conducted a clinical BCI study in an individual with tetraplegia caused by a complete cervical spinal cord injury. This participant was trained to control movement of a computer cursor and a robotic arm using only brain activities captured by an implanted ECoG grid. The ECoG grid offered high spatial and temporal resolution cortical recording, as well as broad coverage over the sensorimotor cortical network. Therefore, we systematically examined the multi-channel ECoG signals recorded during both real-time BCI training and baseline resting period. RESULTS/ANTICIPATED RESULTS: We for the first time identified significant functional connection among cortical patches (<4 mm in diameter) underneath the high-density ECoG grid during BCI learning, which we termed “mesoscale connectivity”. We also identified significant enhancement in cortical activity modulation as the participant improved his performance in BCI control. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings will guide the development of effective BCI training paradigms, and they may also provide additional insights into cortical plasticity for neurorehabilitation purposes.
PROTOTYPING A SENSOR TO MEASURE THE PHYSIOLOGICAL STRESSORS OF AIR AND GROUND MEDICAL TRANSPORT
Reimer AP1,2 1Case Western Reserve University, Cleveland, OH, USA; 2Cleveland Clinic, Cleveland, OH, USA
OBJECTIVES/SPECIFIC AIMS: Air and ground medical transport play a critical role in transferring patients from smaller referring hospitals to large academic medical centers. However, patients who are transferred from one hospital to another hospital experience increased mortality. To better understand how transport affects patient outcomes, it is necessary to develop an in-depth understanding of the extreme environments that patients are subjected to while undergoing medical transport. There is limited knowledge on the affects that the physical stressors of transport have on patients. Therefore, the purpose of this pilot study is to design, build and demonstrate feasibility of a sensor that measures the environmental stressors of medical transport. METHODS/STUDY POPULATION: The sensor unit contains the following sensor elements: a triaxial accelerometer, three gyroscopic sensors, a thermometer, magnetometer, and microphone. Data were recorded from arrival of the transport team to transfer of the patient to the receiving hospital bed. Pilot data were collected on 10 adult patients that underwent either ambulance or helicopter transport. RESULTS/ANTICIPATED RESULTS: The sensor was implemented on 5 ground and 5 helicopter transports. Graphical and analytical analyses of the raw data confirmed reliable measurement across transports. Comparison to other published vibration and noise studies yields similar results supporting measurement validity. Vibration measurements exceeded recommended exposure values. DISCUSSION/SIGNIFICANCE OF IMPACT: Results from this pilot study demonstrate the feasibility of building a new multi-sensor unit that accurately measures the environmental stressors of medical transport, moving beyond commercially available individual sensors. This multi-sensor unit will provide a platform to develop real-time measuring and reporting of the effect of transport on patients.
PUERTO RICAN SPANISH ADAPTATION AND VALIDATION OF THE HEARING HANDICAP INVENTORY FOR THE ELDERLY – SCREENING VERSION
Pintado L3,1, Weinstein BE2, Rodríguez R1, Rivero M4 1Universidad de Puerto Rico, School of Health Professions, San Juan, USA; 2The City University of New York, Graduate Center, New York, NY, USA; 3University of Turabo, School of Health Sciences, Gurabo, USA; 4Universidad de Puerto Rico, School of Nursing, San Juan, USA
OBJECTIVES/SPECIFIC AIMS: Age related hearing loss is the most common type of hearing loss in older persons and it is a disabling problem leading to communication disorders and social dysfunction. The Hearing Handicap Inventory for the Elderly – Screening version (HHIE-S) is designed to be used as a screening tool for hearing loss in older persons. It is well established the need to eliminate the hearing loss health disparity within this population. To the best of our knowledge, no such validated culturally adapted instrument is available for hearing screening in Puerto Rico. This study's objective is to adapt and validate a culturally and linguistically Puerto Rican Spanish version of the HHIE-S. METHODS/STUDY POPULATION: A cultural adaptation of a Spanish version of the HHIE-S will be performed using previously established guidelines to address cultural equivalence. We will perform a pilot test of the final version of the HHIE-S developed for this study. A convenience sample will be obtained from community dwelling older persons. An audiometric hearing screening will be performed for validation purposes. RESULTS/ANTICIPATED RESULTS: N/A DISCUSSION/SIGNIFICANCE OF IMPACT: This project will provide health professionals with an invaluable culturally reliable and valid hearing screening tool for early identification of older persons with unrecognized hearing loss. This work was partially supported by the National Institute on Minority Health and Health Disparities of the NIH under Award Number R25MD007607. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.
QUANTIFICATION OF CERVICAL SPINE MUSCLE FAT: A COMPARISON BETWEEN T1-WEIGHTED AND DIXON BASED MRI
Elliott JM1, Parrish TB2 1Northwestern University, Chicago, IL, USA; 2Northwestern University; Department of Radiology, Feinberg School of Medicine, Department of Radiology, Chicago, IL, USA
OBJECTIVES/SPECIFIC AIMS: To quantify and compare muscle fat percentages (MFP) for the cervical multifidus muscles with T1-weighted imaging and a 3D multi-echo gradient echo (GRE) Dixon based method. METHODS/STUDY POPULATION: 5 asymptomatic participants underwent cervical spine MRI with a Siemens 3 Tesla system. The muscle and fat signal intensities on axial spin-echo T1-weighted images were quantitatively classified for the cervical multifidii from C3-C7, bilaterally. Additional axial GRE Dixon based data for fat and water quantification were used for comparison via paired t-tests. Inter-tester reliability for fat and water measures with GRE images were examined using 1) Pearson's Intra-class correlation coefficient 2) Bland-Altman Plots and 3) Lin's-Concordance Coefficient. P < 0.05 was used to indicate significance. RESULTS/ANTICIPATED RESULTS: Total mean (SD) MFP's (C3-C7) for the multifidii on T1-weighted imaging and GRE were 18.4% (3.3) (range 14–22%) and 18.8% (2.9) (range 15–22%), respectively. The Pearson correlation coefficients for inter-tester reliability on the GRE sequences for the multifidii ranged from .83 – .99, indicating high levels of agreement with segmental MFP measures. Bland-Altman Plots revealed all data points were within 2 SDs and concordance was established between 2 raters, suggesting good agreement for measuring MFP of the healthy cervical multifidii with GRE imaging. DISCUSSION/SIGNIFICANCE OF IMPACT: Results demonstrate reliability between 2 raters of varying experience for MRI analysis of MFP with 3D GRE MRI. The quantification of MFP for healthy cervical musculature is comparable between T1-weighted imaging and 3D GRE. Inclusion of larger samples of symptomatic data (e.g. whiplash) and histological comparison with the reference standard biopsy is warranted.
REGION OF INTEREST VERSUS TRACTOGRAPHY ANALYSIS OF THE CORTICOSPINAL TRACT USING DIFFUSION TENSOR IMAGING
Ontaneda D1, Atthe B1, Sakaie K1, Lin J1, Lowe M1, Phillips M1, Fox R1 1Cleveland Clinic, Cleveland, OH, USA
OBJECTIVES/SPECIFIC AIMS: Several methods exist for analysis of DTI images. We examined the reproducibility and variability of tractography and region of interest (ROI) analysis. METHODS/STUDY POPULATION: Seventeen patients with MS starting natalizumab were imaged monthly for a total of 3 time points. Co-registration of images was conducted with FSL. Regions of interest from normal-appearing white matter in the corticospinal tract were identified and followed longitudinally. Probabilistic tractography was conducted in the corticospinal tract using the ipsilateral cerebral peduncle and hand knob region as target and seed. Average values within the CST region of interest and tract masks were derived for fractional anisotropy (FA), mean diffusivity (MD) longitudinal diffusivity (LD) and transverse diffusivity (TD) using AFNI. Coefficients of variation (CV) were calculated between subjects and between acquisition time points. An F-test was used to test the variance across all time points and subjects. RESULTS/ANTICIPATED RESULTS: Significant differences in variances were seen between ROI and tractography analysis for FA (p = 0.001), MD (p < 0.001), LD (p = 0.001) and TD (p < 0.001) based on F-test statistics. Inter-subject variability ranged between 5.0 and 19.1% and was higher with ROI methodology both in a single acquisition as well as over the different time points for all measures. Inter-acquisition variability was below 1% for both methods and tractography showed a slight advantage over ROI analysis. DISCUSSION/SIGNIFICANCE OF IMPACT: Tractography methods appear to decrease variability and improve reproducibility in DTI measurements. Variability between acquisition time points was minimal (<1%). Variability between subjects was substantial (up to 19.1%) but improved with tractography methods (up to 9.5%).
RIGHT VENTRICULAR STRAIN IN SICKLE CELL-RELATED PULMONARY HYPERTENSION
Desai AA1, Kansal M1, Abbasi T1, Warda A1, Duarte J1, Garcia J1, Machado RF1 1University of Illinois at Chicago, Chicago, IL, USA
OBJECTIVES/SPECIFIC AIMS: An elevated tricuspid valve regurgitation velocity (TRV) on echo and right heart catheterization (RHC)-confirmed pulmonary hypertension (PH) are both independent predictors of mortality in sickle cell disease (SCD). Although 30% have an elevated TRV on echo, only 10% of patients in SCD have RHC-defined PH. We hypothesized that myocardial deformation by speckle-tracking echocardiography (STE) may add to PH detection beyond conventional echocardiography in patients with SCD METHODS/STUDY POPULATION: Standard, STE and RHC data was collected from 25 SCD patients: 8 with no evidence of PH and 17 with PH (mean pulmonary artery pressures, mPAP > = 25 mmHg). RESULTS/ANTICIPATED RESULTS: TRV was only moderately correlated with mPAP (r = 0.38, p < 0.01). In contrast, STE-generated indices such as longitudinal RV peak velocities (cm/sec), strain (%), strain rate (1/sec) of global, lateral and septal walls RV during systole were strongly correlated with mPAP (all r > 0.5, p < 0.001). RV longitudinal global systolic strain rate (LGSR) demonstrated strong correlation with mPAP and an LGSR >-1.22/sec had a 100% sensitivity (55% specificity) to detect PH. RV LGSR had a significant correlation with 6-minute walk distance (r = 0.55, p < 0.001), suggesting prognostic implications for functional status. Over 60 months, 5 patients died all with PH. RV LGSR >-0.87/sec had a 100% sensitivity (95% CI 47–100%) (90% specificity) to predict all-cause mortality. DISCUSSION/SIGNIFICANCE OF IMPACT: Unlike conventional echo, myocardial deformation imaging demonstrates subtle RV systolic dysfunction in SCD. Longitudinal global RV systolic strain rate may be a novel diagnostic test enhancing non-invasive early detection of RHC-defined PH, requiring prospective validation in larger cohorts.
TO WHAT DEGREE DO SUBSTANCE-USE DISORDER COUNSELORS EMPLOY INTERVENTIONS WHICH PROMOTE WELL-BEING?
Krentzman AR1,3, Hassett A2, Mannella K1, Robionson E1, Cranford JA1, Mowbray O1 1University of Michigan Addiction Research Center, Ann Arbor, MI, USA; 2University of Michigan Chronic Pain & Fatigue Research Center, Ann Arbor, MI, USA; 3University of Minnesota School of Social Work, St Paul, MN, USA
OBJECTIVES/SPECIFIC AIMS: Improving the well-being of individuals in treatment for substance use disorders is essential to optimal recovery. A new sober life must be able to compete with the powerful effects of alcohol and other drugs in order to offer a compelling alternative. Substance-use disorder counselors actively work to decrease pathology; however, little is known about the ways in which counselors promote well-being. METHODS/STUDY POPULATION: This pilot study assessed the practice habits of 7 counselors including the frequency with which 24 interventions were employed in the previous week. Twelve interventions decreased pathology (e.g., cravings, relapse, and trauma) and 12 increased well-being (e.g., hobbies, pleasant activities, and gratitude). The percentage of interventions used to increase well-being (PIIWB) was calculated by dividing the frequency of interventions to increase well-being by the frequency of all 24 interventions. Partial correlations (controlling for age and gender) determined associations between PIIWB and other occupational variables. RESULTS/ANTICIPATED RESULTS: PIIWB in this pilot sample ranged from 46% to 64% (M = 53%, SD = 6%). PIIWB was negatively related to the number of years of practice experience (r = –.40) and positively related to the number of hours spent counseling in the previous week (r = .79). DISCUSSION/SIGNIFICANCE OF IMPACT: PIIWB varies among counselors and may be related to other key variables. Future research should determine whether PIIWB is related to job satisfaction and burnout and whether increasing PIIWBs helps alleviate counselor work stress with positive benefits for clients and overall treatment outcomes.
VIRTUAL REALITY GAMES WITH CONTRALATERALLY- CONTROLLED FUNCTIONAL ELECTRICAL STIMULATION FOR MOTOR TRAINING IN HEMIPLEGIA
Fu MJ1, Knutson JS1, Chae J1 1Departments of Electrical Engineering & Computer Science and Physical Medicine & Rehabilitation, Case Western Reserve University, Cleveland, OH, USA
OBJECTIVES/SPECIFIC AIMS: Develop a new intervention for restoring stroke-impaired hand function that uses virtual reality (VR) to motivate motor skill practice and contralaterally-controlled functional electrical stimulation (CCFES) to assist paretic hand movement. Although VR and CCFES therapy have independently shown promise for motor relearning and are complimentary, they have not yet been combined. The first objective is to design VR games with therapists, physiatrists, engineers, and game designers. Each game will then be integrated with robust computer interfaces that translate physical movements into virtual actions. METHODS/STUDY POPULATION: VR+CCFES will be iteratively designed to maximize its clinical relevance and probability for success in future clinical trials. Stroke survivors with hand impairments due to hemiplegia and clinicians will evaluate prototypes of each game and their opinions will be adopted to refine VR+CCFES. RESULTS/ANTICIPATED RESULTS: VR+CCFES will be used in future clinical trials. If improved motor function recovery is demonstrated, then it will be adapted onto portable computers for stroke rehabilitation treatments at home. This will increase motivation and motor practice frequency, which are important for motor relearning. DISCUSSION/SIGNIFICANCE OF IMPACT: Stroke is a leading cause of disability in the US, with incidence of 795,000/yr. – 75% of whom suffer hand function impairments. At 6 mo. post-stroke with conventional care, 65% of survivors are not able to utilize the paretic hand for activities of daily living. Nearly 30% suffer from chronic hemiparesis and are dependent upon caregivers. Thus, novel methods such as this work are needed to improve the standard of care for restoring motor function to stroke survivors suffering from hemiplegia.
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ANALYSIS OF UTILIZATION OF ABLATION PROCEDURES FOR CARDIAC ARRHYTHMIAS IN THE UNITED STATES FROM 2000–2008
Pant S1, Deshmukh A1, Mehta K4, Badheka AO2, Kumar G3, Mehta J1 1UAMS, Little Rock, AR, USA; 2Miller School of Medicine-University of Miami, Miami, FL, USA; 3Medical College of Wisconsin, Milwaukee, WI, USA; 4Drexel University, Philadelphia, PA, USA
OBJECTIVES/SPECIFIC AIMS: To evaluate the utilization of radiofrequency ablation for various arrhythmias throughout the United States from 2000–2008. METHODS/STUDY POPULATION: Using the Nationwide Inpatient Sample database, we identified atrial fibrillation (AFIB), atrial flutter (AFL), PSVT and VT patients treated with ablation. We investigated the trends, in-hospital mortality and total cost of care for each hospitalization. RESULTS/ANTICIPATED RESULTS: A total of 47,259 of AFIB, 38,881 AFL, 37,227 PSVT, 13,942 VT ablations were performed from 2000 to 2008. We noted an overall increase in the frequency of these procedures performed over this period (Figure 1). The average increase for AFIB, AFL, PSVT and VT was 46%, 12.7%, 4.6% and 5.5% respectively. The overall in-hospital mortality over this period remained nearly stable for AFIB, AFL and PSVT ablation. Although, relative increase for AFIB, AFL and PSVT ablation was 0.2%, 0.1%, and 0.1%, respectively (all p-NS), the in-hospital mortality for VT ablation increased substantially and significantly over the study period (1.6%, p < 0.01) (Figure 2). The total cost of care for ablation procedures increased likewise for all arrhythmias in a parallel fashion. Total hospitalization cost was highest for the AFIB for most of the years, followed by VT, AFL and PSVT (Figure 3) except for 2008 where VT ablation cost superseded the rest. DISCUSSION/SIGNIFICANCE OF IMPACT: The utilization of ablation for various arrhythmias has significantly increased from 2000–2008, most remarkably for AFIB. The in-hospital mortality has not changed over the years except for VT ablation where it increased. Cost of care increased for all arrhythmias in parallel, AFIB ablation cost being higher in general followed by VT, AFL and PSVT.
ASSESSING THE LITERATURE EVIDENCE FOR LABELED ADVERSE DRUG REACTIONS
Duke J1, Cadwallader J1 1Indiana University, Indianapolis, IN, USA
OBJECTIVES/SPECIFIC AIMS: A well-recognized issue in drug product labeling (i.e., package inserts) is the problem of overwarning, in which long lists of potential adverse reactions lead to cognitive overload for providers. One potential approach to addressing overwarning is to use various filters to highlight or de-emphasize warnings based on clinical importance. One such marker of clinical importance may be the supporting literature. In this study, we perform a large scale assessment of the available literature for all labeled adverse drug reactions (ADRs). METHODS/STUDY POPULATION: Using a database of 131,857 drug-adverse reaction pairs, we performed a series of queries using the PubMed Entrez Programming Utilities. For each drug-reaction pair, we performed the following searches: 1) “[drug name] [ADR name]”; 2) “[drug name] [ADR name] /adverse effect”; and 3) “[drug name]/adverse effect [ADR name]”. We then performed descriptive statistics regarding the number of citations for each drug-reaction pair by individual drugs, across drug classes, and across all drug labels. RESULTS/ANTICIPATED RESULTS: Using the most liberal search strategy, 53% of labeled ADRs showed at least one publication associating the drug and adverse reaction. The most conservative query (requiring MeSH reference to an adverse effect) resulted in just 41% of ADRs meeting this criterion. Relatively few labeled ADRs has 'strong' literature support, with 21% having ≥3 citations and 12% having ≥10 citations. DISCUSSION/SIGNIFICANCE OF IMPACT: A substantial proportion of adverse reactions appearing on drug labels have no corroborating evidence in the medical literature. Filtering labels to highlight highly studied reactions while de-emphasizing those with no evidence outside the labeling my help reduce information overload and increase the clinical utility of this important information resource.
BASELINE CHARACTERISTICS OF TRANSGENDER YOUTH NAÏVE TO CROSS SEX HORMONE THERAPY
Olson J1,2, Schrager S1, Belzer M1,2, Simons L1, Clark L1,2 1Children's Hospital Los Angeles, Los Angeles, CA, USA; 2Keck School of Medicine at USC, Los Angeles, CA, USA
OBJECTIVES/SPECIFIC AIMS: Standard of care for transgender youth includes cross sex hormone therapy, however evidence for efficacy is lacking. This abstract presents baseline data from an ongoing observational study examining the impact of hormone treatment on transgender youth who are naïve to cross sex hormone therapy. METHODS/STUDY POPULATION: Transgender youth (aged 12 to 24) completed a computer assisted survey that assessed gender dysphoria, depression, sexual behavior and suicidality. Physiologic data collected prior to starting hormones was abstracted from medical charts. RESULTS/ANTICIPATED RESULTS: 70 participants were evaluated for baseline physiologic, 66 for psychosocial parameters. 50% were assigned a female gender at birth. 34 (51.5%) were Caucasian, 28.8% Latino/a, 10.6% African American, and 9.1% other. Blood pressure, liver function tests, lipid and hormone levels were within normal clinical range for most of the participants. Youth discovered their internal gender was different from their assigned gender at ages ranging from 2 to 22 years (mean 7.9 y; SD 4.4), and disclosed to their family at ages ranging from 3 to 23 years (mean 16.8; SD 4.1). Gender dysphoria was high overall, but was significantly higher among transgender males. Five youth (8.1%) had Beck Depression scores in the severe or extreme range. 30% had made at least one suicide attempt. Of the sexually active youth (6) 18.8% had traded sexual activity for survival. DISCUSSION/SIGNIFICANCE OF IMPACT: Transgender youth are aware of the incongruence between their internal gender identity and their assigned gender at early ages. Levels of depression and suicidality demonstrate that youth need timely and appropriate intervention. Evaluation of these youth over time will help determine the impact of cross sex hormones and mental health therapy.
BETTER IN-HOSPITAL SURVIVAL WITH PERCUTANEOUS ENDOSCOPIC GASTROSTOMY (PEG) THAN WITHOUT
Arora G1, Rockey D1, Gupta S1 1UT Southwestern, Dallas, TX, USA
OBJECTIVES/SPECIFIC AIMS: Though PEG is associated with a high short-term mortality, that attributable to the procedure itself is estimated at 0.5%. However, this is difficult to accurately assess as no study reporting outcomes after PEG has ever included a control group. We hypothesized that PEG-attributable mortality is higher and performed propensity-score matching (PSM) to derive controls to test this hypothesis. METHODS/STUDY POPULATION: We identified all PEGs in the nationally representative Nationwide Inpatient Sample (NIS) dataset. For PSM, we included common indications for PEG (malnutrition, dysphagia, stroke, dementia, head trauma, head and neck cancer); factors associated with PEG: age, gender, length of hospital stay, admission source, discharge disposition; and hospital characteristics (teaching status, location, control and bed size). Unique controls (1:1) that were equally likely to get a PEG but did not, were derived. In-hospital mortality was compared between the 2 groups using conditional logistic regression. Independent validation of results was done using 2007 NIS data. Sensitivity analysis was performed by incorporating known predictors of mortality (KPM) after PEG such as weight loss, COPD, CHF, etc. within PSM (Am J Gastroenterol. 2010; 105 (S1): S402) and by adjusting for Charlson index. RESULTS/ANTICIPATED RESULTS: 143,082 of 181,196 patients who underwent PEG were matched. Both cohorts were well-matched on variables used for PSM and KPM. Contrary to our hypothesis, we found that the in-hospital mortality was lower in those who underwent PEG (10.9% vs. 11.7%; OR 0.92 [95% CI 0.87–0.97]; NNT = 125). Sensitivity analyses and independent validation confirmed the robustness of these results. DISCUSSION/SIGNIFICANCE OF IMPACT: In-hospital mortality after PEG is high. However, compared to a control group matched on indications for PEG and important comorbidities, PEG was associated with improved in-hospital survival.
EFFECTIVENESS OF COUPLE THERAPY: SITUATIONAL COUPLE VIOLENCE & NEUROENDOCRINOLOGY
Karakurt G1 1Case Western Reserve University, Cleveland, OH, USA
OBJECTIVES/SPECIFIC AIMS: This project aims to explain whether couple therapy can be effective in preventing the escalation and recurrence of situational couple violence. METHODS/STUDY POPULATION: Participants of this study will consist of 30 couples who are currently in a relationship. Among these couples, 15 will be placed in a control group, and 15 will be placed in an intervention group. The intervention group will be entered into 8 to 12 weeks of couple therapy. Therapy sessions will last approximately 60 minutes and will occur on a weekly basis. Data will be collected at three time points: pre-test, post-test, and follow-up. Both behavioral and physiological measurements will be used to evaluate the effectiveness of conjoint therapy. The change in violent behaviors will be examined along with changes in HPA axis and PNS activity, as well as questionnaires measuring attachment patterns, affect regulation, dyadic adjustment, emotional abuse, and intimate partner violence. RESULTS/ANTICIPATED RESULTS: We are expecting to find that couple therapy will gradually promote affect regulation of both partners in orchestration; i.e., improvement in one partners' ability to regulate emotions will have a positive effect in the other partner's ability to regulate emotions. DISCUSSION/SIGNIFICANCE OF IMPACT: Findings of this study will provide biologically interpretable and quantitative grounds for the effectiveness of couple therapy and pave the way for more comprehensive and large-scale effectiveness studies, which, in turn, may establish couple therapy as a strategy that will complement 'parallel track' (offering therapy and educational programs to male offenders and female victims separately) in preventing and diminishing IPV.
EFFECTS OF DEEP BRAIN STIMULATION ON GAIT AND BALANCE IN PARKINSON'S DISEASE
Wagle Shukla A1, Defranco M1, Okun M1 1University of Florida, Gainesville, FL, USA
OBJECTIVES/SPECIFIC AIMS: To determine the effects of Deep Brain Stimulation (DBS) on gait and balance in Parkinson's Disease (PD) measured quantitatively and longitudinally. METHODS/STUDY POPULATION: PD subjects undergoing bilateral DBS surgery either in subthalamic nucleus (STN) or globus pallidus (GPi) were enrolled. Demographics and Unified Parkinson's disease rating Scale (UPDRS) scores were recorded. Gait and balance was recorded using 10 m walk test, Berg Balance scale and Timed up and Go test before and two+ months after surgery. All recordings were ON medication and postoperatively ON medication and ON stimulation. Wilcoxon-signed-rank test was used for pre and postoperative comparisons. RESULTS/ANTICIPATED RESULTS: 15 patients were enrolled. Eight underwent bilateral STN DBS (7 males, 1 female; mean age 65 years; mean UPDRS score 26). Seven underwent GPi DBS (5 males, 2 females; mean age 64.5 years; mean UPDRS score 25). Statistical testing was negative however on individual analyses, one patient revealed significant postoperative deterioration in the Berg Balance scale for both STN (score preoperative 50 and postoperative 38) and GPi stimulation (score preoperative 50 and postoperative 38). For Timed Up and Go test one patient with STN stimulation demonstrated worsening (10 seconds preoperatively to 18 seconds postoperatively). 10 m walk results were unremarkable. DISCUSSION/SIGNIFICANCE OF IMPACT: DBS is a well-established treatment for PD. However there are increasing concerns over its adverse effects on gait and balance. It is not clear if these effects differ by the target of stimulation (subthalamic nucleus or globus pallidus). In this small cohort, the effects didnot differ based on the target of stimulation. A larger sample with longer follow-up is planned for further elucidation.
IMPACT OF TRANSCUTANEOUS CARBON DIOXIDE MONITORING ON NEONATAL BLOOD GAS MEASUREMENT
Mukhopadhyay S1,2, Puopolo KM2,3 1Childrens Hospital Boston, Boston, MA, USA; 2Brigham and Womens Hospital, Boston, MA, USA; 3Harvard Medical School, Boston, MA, USA
OBJECTIVES/SPECIFIC AIMS: (1) To compare total number of blood gases (BG) obtained between comparable intubated infants in the Neonatal Intensive Care Unit (NICU) managed with and without TCOM (2) To measure correlation between TCOM and BG pCO2 measurements. METHODS/STUDY POPULATION: TCOM technology was introduced for clinical use in our NICU in 10/2010. This is a retrospective time series study. Infants intubated for >48 hours in a 'preTCOM' period (11/2009–9/2010) were compared with infants matched by gestational age (GA) and birth weight (BW) in a 'postTCOM' period (11/2010–9/2011), who had TCOM measurements for ≥ 50% of their ventilated time. Outcome measures: (a) frequency of BG/days of intubation, and (b) mean difference of simultaneous BG and TCOM measurements. RESULTS/ANTICIPATED RESULTS: 34 infants met study criteria. There were no significant differences in GA, BW, use of antenatal steroids, duration of intubation and hospitalization. Mean number of BG/day of intubation was 4.1 (pre-TCOM) and 3.2 (post-TCOM) (p = 0.1). For infants in the post-TCOM period, the mean difference (pCO2-TCOM measure) was 6.9 (± 10.1) with 50.3% measures varying ≤7.5 mm Hg. DISCUSSION/SIGNIFICANCE OF IMPACT: Optimal care of infants requiring mechanical ventilation in the NICU requires monitoring of oxygenation and ventilation. Unlike non-invasive oximetry for oxygenation, assessment of ventilation requires direct pCO2 measurement from blood. TCOM offers a non-invasive, continuous assessment of ventilation. The impact of this technology on the care of NICU infants remains unclear due to concern about TCOM measurement accuracy. In the preliminary results of our study TCOM pCO2 correlated 50% of the time with BG pCO2. We found no significant difference in the frequency of BG measurement among ventilated infants monitored with TCOM.
INTENSITY MODULATED RADIATION THERAPY RESULTS IN SIMILAR SURVIVAL TO 3D-CONFORMAL RADIATION FOR STAGE III NON-SMALL-CELL LUNG CANCER
Harris JP1, Loo BW1,2, Diehn M1,2 1Stanford University - Department of Radiation Oncology, Stanford, CA, USA; 2Stanford Cancer Institute, Stanford, CA, USA
OBJECTIVES/SPECIFIC AIMS: For stage III non-small-cell lung cancer (NSCLC) the most commonly used radiation technique is 3-dimensional conformal radiation therapy (3D-CRT), although intensity modulated radiation therapy (IMRT) is being increasingly employed. There are concerns that breathing-associated tumor motion may decrease the radiation dose to portions of a tumor when using IMRT, which could potentially lead to inferior clinical outcomes. No randomized studies have compared these treatments. Thus, we performed a population-based comparative effectiveness analysis of radiation treatment strategies for stage III NSCLC. METHODS/STUDY POPULATION: We used Surveillance, Epidemiology, and End Results (SEER)-Medicare data to identify 5,846 patients aged 65 or older with stage III NSCLC from 2002–2007, whose treatment included radiotherapy. Using Cox regression and propensity score matching, we compared IMRT, 3D-CRT, and 2D based planning and simulation (2D-CRT) with regards to overall and cancer specific survival (OS, CSS). RESULTS/ANTICIPATED RESULTS: The proportion of patients treated with IMRT increased from 1.8% in 2002 to 13% in 2007, while use of 2D-CRT decreased from 33% to 6%. In multivariate analysis, IMRT was associated with similar OS (hazard ratio [HR] 0.96, 95% confidence interval [CI] 0.85–1.09) and CSS (HR 0.92, 95% CI 0.80–1.06) compared to 3D-CRT, and results were similar in propensity score matched analysis. Superior OS and CSS were associated with IMRT (OS HR 0.72, 95% CI 0.60–0.86; CSS HR 0.68, 95% CI 0.56–0.82) and 3D-CRT (OS HR 0.86, 95% CI 0.79–0.95; CSS HR 0.86, 95% CI 0.78–0.95) compared to propensity score matched 2D-CRT cohorts. DISCUSSION/SIGNIFICANCE OF IMPACT: For stage III NSCLC, IMRT results in similar overall and cancer-specific survival to 3D-CRT, and both are superior to 2D-CRT.
LONG-TERM DIET AND EXERCISE OUTCOMES AFTER CORONARY ARTERY BYPASS SURGERY: A REVIEW OF RECENT LITERATURE
Coyan G1, Reeder KM2, Vacek JL3 1University of Kansas Medical Center, Kansas City, KS, USA; 2Goldfarb School of Nursing at Barnes-Jewish College, St. Louis, MO, USA; 3Mid-America Cardiology, Kansas City, KS, USA
OBJECTIVES/SPECIFIC AIMS: The purpose of this review of recent extant literature was to explore and evaluate evidence on long-term CABG diet and exercise outcomes. METHODS/STUDY POPULATION: Literature search methods were conducted using the Medline computerized data base. Search terms included diet, exercise, and coronary artery bypass surgery. The search was limited to English language articles published between 1995 and 2013. Reference lists from relevant articles were culled. Of over 400 abstracts reviewed, only studies that addressed long-term outcomes were retained for this review. RESULTS/ANTICIPATED RESULTS: The limited available literature indicates exercise positively impacted psychosocial well-being and physical fitness. Current evidence indicates diet and exercise interventions are effective short-term, but intervention effects fade over time. Potential age and gender differences were found across studies reviewed; however, further research is needed with more rigorous designs to replicate and confirm findings. DISCUSSION/SIGNIFICANCE OF IMPACT: Additional research is warranted on long-term outcomes of lifestyle changes and self-management of chronic coronary heart disease. Studying long-term outcomes of CABG surgery will contribute evidence to clinical practice guidelines.
LONG-TERM OUTCOME OF DAILY NASAL IRRIGATION FOR THE TREATMENT OF PEDIATRIC CHRONIC RHINOSINUSITIS
Pham V1, Sykes K2, Wei J2 1University of Kansas School of Medicine, Kansas City, KS, USA; 2Department of Otolayrngology-Head and Neck Surgery, University of Kansas School of Medicine and Medical Center, Kansas City, KS, USA
OBJECTIVES/SPECIFIC AIMS: Chronic rhinosinusitis (CRS) results in significant morbidity and healthcare expenditure. This study reviewed characteristics of a cohort of children with confirmed diagnosis of CRS based on history and computed tomography who have failed medical therapies, utilization of nasal irrigation for subsequent symptoms after initial experience using nasal irrigation, and long-term outcome using nasal irrigation. METHODS/STUDY POPULATION: Review and survey of 144 pediatric patients diagnosed with CRS between July 2003 and January 2012. RESULTS/ANTICIPATED RESULTS: One hundred five patients were included in our study. Mean age was 8.0 years, and 65.7% were male. Presenting symptoms included congestion (95%), cough (80%), rhinorrhea (61%), headache (49%), and fatigue (40%). Comorbid conditions include positive allergy test (66%), asthma (57%), and GERD (28%). After 6 weeks of irrigation, 58.8% of patients reported resolution of symptoms. Lund-Mackay scores averaged reductions of 4.14 and 4.38 on the left and right sides, respectively. Patients requiring FESS were, on average, 3.6 years of age older than those that did not receive FESS (p = 0.0005). Of the 34 patients contacted, 50.0% have used irrigated in the last 12 months, and 7 received sinus surgery. Median length of follow up was 46 months (range 15–104). There were no significant differences in age, Lund-MacKay score changes, and symptom resolution proportions between those contacted and those that were not contacted. DISCUSSION/SIGNIFICANCE OF IMPACT: Nasal irrigation is effective as a first line treatment for pediatric CRS as well as for recurring symptoms. Children with symptoms and diagnosis of CRS should always be encouraged to use once daily nasal irrigation and may avoid elective endoscopic sinus surgery.
NEONATAL WHITE MATTER DEVELOPMENT IN VERY PRETERM INFANTS AND DEVELOPMENTAL OUTCOME AT AGE 2
Rogers C1, Smyser T1, Inder T1, Neil J1 1Washington University School of Medicine, Saint Louis, MO, USA
OBJECTIVES/SPECIFIC AIMS: Preterm infants are at greater risk of poor developmental outcomes. This study aims to assess if alterations in regional white matter development measured with Diffusion Tensor MRI (DTI) at term-equivalent are related developmental outcomes at age 2 in a cohort of very preterm infants. METHODS/STUDY POPULATION: Infants born <30 weeks gestational age (GA) had DTI performed at term-equivalent (36–42 weeks GA). Regions of interest were manually placed for DTI measures of fractional anisotropy (FA) and mean dIffusivity (MD). Children were assessed at age 2 years with the Modified-Checklist for Autism in Toddlers (M-CHAT), the Infant Toddler Social Emotional Assessment (ITSEA) and the Bayley Scales of Infant Development III (Bayley III). RESULTS/ANTICIPATED RESULTS: Preliminary results from 62 VPT children with DTI and age 2 outcomes indicated that altered FA in the right anterior limb of the internal capsule (ALIC) and left ALIC MD is associated with worse Bayley III Cognitive (p = .04), Language (p = .045) and Fine Motor (p = .01) scores. Alterd right ALIC FA is also related to positive autism screening (p = .01). Lower Bayley III Gross Motor scores were associated with lower FA in the right centrum semiovale (p = .002) and posterior limb of the internal capsule (PLIC) (p = .03). Lower FA in the right PLIC was also related to lower Fine Motor scores (p = .04). Lower ITSEA Competence scores were associated with altered left ALIC FA (p = .006) and left cingulum MD (p = .02). DISCUSSION/SIGNIFICANCE OF IMPACT: Alterations in regional white matter detectable during the neonatal period are related to cognitive, language, motor, and social-emotional development at age 2 in VPT infants. These findings suggest that VPT children at-risk for poor development may be identified prior to discharge for targeted surveillance of specific deficits potentially leading to more effective early intervention.
NURSE WORKLOAD, MISSED CARE, AND INFANT OUTCOMES IN THE NEONATAL INTENSIVE CARE UNIT
Tubbs Cooley HL1,3, Pickler RH1,2, Mark BA4, Kotagal U1,3, Carle AC1,3, Solti I1,3 1Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA; 2The Ohio State University, Columbus, OH, USA; 3University of Cincinnati, Cincinnati, OH, USA; 4University of North Carolina at Chapel Hill, Chapel Hill, NC, USA
OBJECTIVES/SPECIFIC AIMS: Missed nursing care is an often theorized mediator of the effect of nurse workload on patient outcomes in the hospital setting, though this relationship has not yet been explicitly tested. The primary aim of this study is to examine the role of missed nursing care as a contributor to infant outcomes both independently and as a mediator of the effect of nurse workload on outcomes. METHODS/STUDY POPULATION: This is a prospective observational study occurring in a single NICU over 12 months. We are actively enrolling NICU nurses, who will provide information during data collection cycles on workloads and missed care specific to each baby cared for during a shift. Infants will be linked to nurse reports and infant outcome data will be extracted from the electronic health record. We will construct a large, de-identified database from these data sources and will construct path analysis and structural equation models. RESULTS/ANTICIPATED RESULTS: Based on our pilot work, we expect to enroll approximately 80% (n = 100) of nurses working in the NICU. We expect to confirm our primary hypothesis that instances of missed care partially explain the effects of nurse workload on patient outcomes. DISCUSSION/SIGNIFICANCE OF IMPACT: The proposed work is significant because the outcomes under investigation are largely preventable when evidence-based care processes, such as bundles or feeding protocols, are reliably implemented by front-line nurses. Our innovative approach will refine existing knowledge about the relationships between nursing care and patient outcomes, resulting in a broadened conceptualization of systems contributors to outcomes and identification of areas for future intervention work.
NURSE-LED GROUP CLINIC INTERVENTION REDUCED DEPRESSION AMONG AFRICAN AMERICAN PATIENTS WITH HEART FAILURE
Piamjariyakul U1, Smith CE1, Wick J1, Fitzgerald S1 1University of Kansas Medical Center, Kansas City, KS, USA
OBJECTIVES/SPECIFIC AIMS: To examine the effect of nurse-led group clinic appointment discussion on depression of patients with heart failure (HF) by race. The Chronic Care Model was used to guide the study. METHODS/STUDY POPULATION: The sample was comprised of 155 patients (44.5% were African American) with HF who were in a randomized clinical trial; 44% (n = 68) attended a nurse-led group discussion (intervention group) and 56% (n = 87) were standard care (routine HF education). The CESD (>20 cut-of score) questionnaires were used to screen for depression. RESULTS/ANTICIPATED RESULTS: There was no significant difference on depression scores between racial or treatment groups at baseline. A two-way ANOVA was conducted to test the effect of race (African American vs White) and treatment group (intervention vs control) on the change in depression score from baseline to 6-month post intervention. Levene's test indicated homogeneity of variance between groups. There was a significant interaction between the race and treatment group on the change of depression score, F (1, 151) = 4.68, p = .032. Fisher LSD post-hoc analysis indicated the improvement on depression on all patients, but significant greater improvement on depression score was found among African American patients in the intervention group (Mean difference = 3.2, p < .05). In the group discussion, patients readily shared emotions and encouraged each other to share their feelings and support needs with their family members and friends as a way of managing stress. DISCUSSION/SIGNIFICANCE OF IMPACT: Patients with HF, who have clinical depression, are at greater risk for re-hospitalization and death. Patients benefit from nurse-led multidisciplinary group discussions. The findings will guide the development of intervention program to improve depression among patients with HF and other chronic illnesses.
OUTCOMES OF ELDERLY STAGE I LUNG CANCER PATIENTS TREATED WITH SEGMENTECTOMY VIA VIDEO ASSISTED THORACOSCOPIC SURGERY OR OPEN RESECTION
Smith C1, Wisnivesky JP1 1Mount Sinai School of Medicine, NY, NY, USA
OBJECTIVES/SPECIFIC AIMS: Video-assisted thoracic surgery (VATS) is considered an alternative to open lobectomy for the treatment of non-small cell lung cancer (NSCLC). Limited data is available however, regarding the equivalence of open vs. VATS segmental resections, particularly among elderly patients. In this study, we used population-based data to compare postoperative and oncologic outcomes following open vs. VATS segmentectomy for early NSCLC. METHODS/STUDY POPULATION: We identified all stage I NSCLC patients >65 year treated with VATS or open segmentectomy from the Surveillance, Epidemiology, and End Results registry linked to Medicare claims. We used propensity score methods to control for differences in the baseline characteristics of patients treated with VATS vs. open segmentectomy. Study outcomes included perioperative complications, need for intensive care unit (ICU) admission, extended length of stay, perioperative mortality as well as overall and lung cancer-specific survival. RESULTS/ANTICIPATED RESULTS: Of the 577 study patients, 27% underwent VATS resection. VATS-treated patients had lower rates of postoperative complications (odds ratio [OR]: 0.55, 95% confidence interval [CI]: 0.37–0.83), need for ICU admission (OR: 0.18, 95% CI: 0.12–0.28), and decreased length of stay (OR: 0.41, 95% CI: 0.41–0.81) after adjusting for propensity scores. Postoperative outcomes were not significantly different across groups after adjusting for surgeon characteristics. Overall (hazard ratio [HR]: 0.80, 95% CI: 0.60–1.06) and lung cancer-specific (HR: 0.71, 95% CI: 0.45–1.12) survival was similar among patients treated with VATS vs. open segmentectomy. DISCUSSION/SIGNIFICANCE OF IMPACT: VATS segmentectomy can be safely performed among elderly patients with early stage NSCLC and is associated with equivalent postoperative and long-term oncologic outcomes.
PLATELET-DERIVED MICROPARTICLE GENERATION OF ECMO SYSTEMS AT SIMULATED FLOW RATES FOR NEONATES
Meyer AD1,3, Wiles AA2, Rivera O3, Rais-Bahrami K3, Freishtat RJ2 1U.T. Health Science Center at San Antonio, San Antonio, TX, USA; 2Children's National Research Institute, Washington, DC, USA; 3Children's National Medical Center, Washington, DC, USA
OBJECTIVES/SPECIFIC AIMS: Platelets are the most commonly transfused blood product during extracorporeal membrane oxygenation (ECMO). ECMO is a miniature form of cardiopulmonary bypass (CPB) that can be a life-saving treatment for infants with heart or lung failure. Research has shown that CPB activates platelets to release platelet-derived microparticles (PMP), a mediator for increasing thrombogenicity and inflammation. Few if any studies have examined the generation of PMPs in a neonatal ECMO system. The objective of this study was to examine the generation of PMPs in a variety of pediatric ECMO systems. METHODS/STUDY POPULATION: Three newer components (Jostra Rotaflow centrifugal pump, Quadrox-D and Quadrox-iD Pediatric Oxygenator, Maquet) were compared to the standard roller-head pump (Jostra, Maquet) and silicone membrane oxygenator (0800, Medtronic) in a simulated in vitro ECMO circuit circulating whole swine blood at 300 ml/min. Five circuit combinations were examined for hemolysis and microparticle generation during 6 hrs of continuous use. Hemolysis was measured by plasma free hemoglobin (fPH) using a spectrophotometer. Microparticle generation was measured using flow cytometry. RESULTS/ANTICIPATED RESULTS: All of the ECMO systems created fPH at a similar rate as compared to the static. The circuits with a centrifugal pump had a significantly increased microparticle event rate compared to the roller pump; 5.64 events/min vs. 0.88 events/min (p < 0.001), respectively. DISCUSSION/SIGNIFICANCE OF IMPACT: In a low-flow neonatal environment, centrifugal pumps generate more microparticles when compared to roller-head pumps. Further study examining if the generation of microparticles during ECMO will predict patient outcomes is needed.
PNEUMOCOCCAL CONJUGATE VACCINE-13 IMPACT ON OTITIS MEDIA IN YOUNG CHILDREN
Marom T1, Tan A1, Pierson KS1, Wilkinson GS1, Freeman JL1, Chonmaitree T1 1University of Texas Medical Center, Galveston, TX, USA
OBJECTIVES/SPECIFIC AIMS: Streptococcus pneumoniae (Sp) is the predominant otitis media (OM) pathogen. An effective vaccination against Sp can substantially reduce OM incidence. The 13-valent pneumococcal conjugate vaccine (PCV13) offers a broader protection than the previously used PCV7. We sought to determine the impact of PCV13 on OM visit rates in young children. METHODS/STUDY POPULATION: We identified OM cases in children aged≤6 years during 2005–2011 in the ClinInformatics database, which consists claims from a large private managed U.S. health care plan. OM visit rates in the years before (2005–2009), during (2010) and after (2011) PCV13 licensure were compared for children <2 years (PCV13 eligible) and children aged 2–6 years (not routinely given PCV13).OM visits rate ratios for the two age groups were calculated. The effect of PCV13 was estimated by comparing differences in the rate ratios for these two groups during pre- and post- PCV13 introduction periods. RESULTS/ANTICIPATED RESULTS: The number of children aged ≤ 6 years ranged between 610,860 and 713,937 per year during 2005–2011; 26.8% of the subjects were younger than 2 years. Documented immunization rates for PCV7 and/or PCV13 vaccines in children ≤2 years were 86% – 88% (2005–2009); 88% (2010) and 89% (2011). During pre-PCV13 years (2005–2009), the PCV13 introduction year (2010) and the post-market year (2011), OM visit rates per child-year in children aged <2 were 1.92, 1.65 and 1.12, compared with rates of 1.36, 1.22 and 1.08 in children aged 2–6 years, respectively. The rate ratios between children aged <2 and 2–6 years declined by 4.7% in 2010 and 26.6% in 2011 from pre-PCV13 years. There were no appreciable changes in OM visit rates in children aged 2–6 years during the studied years. DISCUSSION/SIGNIFICANCE OF IMPACT: PCV13 introduction is associated with a sharp decline in OM visits rates in young children.
PREGNANCY OUTCOMES IN RENAL TRANSPLANTS WITH AND WITHOUT LUPUS
Broder A1 1Albert Einstein College of Medicine/Montefiore Medical Center, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: The majority of individuals with lupus (SLE) are young women of child-bearing age. Ten to 30% of individuals with lupus nephritis progress to end stage renal failure (ESRF) within 10 years of diagnosis. Pregnancy outcomes in renal transplants with SLE have not been studies previously. The objective of this study was to compare pregnancy outcomes and complications in SLE and non-SLE using the Unites States Renal Data System (USRDS), an NIH-sponsored data collection system that collects information about ESRF incidence, prevalence, patient characteristics, and treatment modalities. METHODS/STUDY POPULATION: Women who received renal transplant between 20 and 40 years of age were identified from the USRDS 2005–20011. Pregnancy complications and outcomes that occurred after renal transplant were identified from the claims files using ICD9 diagnostic codes (631–674). RESULTS/ANTICIPATED RESULTS: There were 4265 women with SLE and 52279 women without SLE who met inclusion criteria. Pregnancy related ICD9 codes were reported in 107 (2.5%) SLE and 797 (1.5%) non-SLE individuals. The rates of normal full-term pregnancies were 1.3% in non-SLE and 1.2% in SLE, p = 0.044. The rates of mild hypertension/pre-eclampsia were 10.1% in non-SLE and 8.8% in SLE (p = 0.051). The rates of severe pre-eclampsia were 3.7% in non-SLE and 4.4% in SLE (p = 0.57). There were no statistical differences in the rates of ectopic pregnancies, spontaneous abortions, premature labor, fetal malformations, and thombotic complications post-partum. DISCUSSION/SIGNIFICANCE OF IMPACT: In this bivariate analysis pregnancy and complication rates were similar between renal transplants with and without SLE. Future analysis will be conducted to determine variables associated with unfavorable pregnancy outcomes in these individuals.
PREOPERATIVE BRAIN MRI IN CONGENITAL HEART DISEASE INFANTS IS ASSOCIATED WITH OUTCOME
Ortinau C1, Beca J2, Neil J1, Shimony J1, Alexopoulos D1, Inder T1 1Washington University in St. Louis School of Medicine, St. Louis, MO, USA; 2Starship Children's Hosital, Auckland, New Zealand
OBJECTIVES/SPECIFIC AIMS: Determine the association of preoperative brain size and microstructure with outcome at two years of age. METHODS/STUDY POPULATION: Sixty-seven infants with complex congenital heart disease (CHD) were enrolled in a prospective longitudinal study and underwent preoperative brain MRI. Brain metrics, a simple measurement technique of brain size, was applied across multiple regions of the brain. Diffusion tensor imaging, a technique used to quantify microstructural development, was applied in 19 infants. The apparent diffusion coefficient (ADC) and fractional anisotropy were measured in 14 regions. Infants were followed-up at two years of age and underwent the Bayley Scales of Infant Development-Third Edition (BSID-III) to assess motor outcome with the Psychomotor Developmental Index (PDI) and cognitive outcome with the Mental Developmental Index (MDI). RESULTS/ANTICIPATED RESULTS: Nine infants died and three were lost to follow-up, leaving 55 infants who underwent the BSID-III. There was an association between smaller brainstem size and lower PDI, even after controlling for postmenstrual age at MRI, head circumference, and sex (r = 0.337, p < 0.05). There was no association for other brain metrics. Controlling for the same factors, there was an association between higher ADC values in the left (r = –0.60, p < 0.05) and right (r = -0.58, p < 0.05) posterior limb of the internal capsule and lower MDI. DISCUSSION/SIGNIFICANCE OF IMPACT: Infants with CHD have altered brain growth and microstructure preoperatively. Our data demonstrate an association of brainstem size with motor outcome and microstructural development with cognitive outcome. These data support the need for further studies evaluating the impact of brain development on outcome to guide the utility of preoperative MRI in the clinical setting.
PROGNOSIS IN HIV INFECTED PATIENTS WITH NON-SMALL CELL LUNG CANCER
Sigel KM1, Crothers K2, Dubrow R3, Sigel C4, Wisnivesky J1 1Mount Sinai School of Medicine, New York, NY, USA; 2University of Washington School of Medicine, Seattle, WA, USA; 3Yale School of Public Health, New Haven, CT, USA; 4Memorial Sloan-Kettering Hospital, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: HIV+ individuals are at greater risk of developing lung cancer than the general population. It is unclear whether lung cancer prognosis is altered in the setting of HIV infection. Using population-based data, we evaluated whether HIV infection was associated with worse prognosis in patients with non-small cell lung cancer (NSCLC). METHODS/STUDY POPULATION: We performed a cohort study using the SEER registry linked to Medicare claims from 1996–2007. Using these data we identified 267 HIV+ patients with NSCLC. These cases were matched by age, gender, race, and reason for Medicare entitlement to 1208 patients with no evidence of HIV infection. Cause of death was classified as lung cancer related or non-cancer related using SEER death data. The association between HIV status and lung cancer survival was evaluated using Kaplan-Meier curves, conditional probability function (CPF) methods to account for competing risks of death. RESULTS/ANTICIPATED RESULTS: Cancer stage at presentation (p = 0.6) and histology (p = 0.6) did not differ by HIV status. Overall survival was worse among HIV+ NSCLC patients compared to patients without evidence of HIV (7 mo. vs. 25 mo.; p < .001). Using competing risks methods (CPF), we found that HIV+ patients had poorer NSCLC prognosis (p < .001). In a multivariable CPF regression, HIV was associated with a greater risk of lung cancer death (odds ratio: 1.7, p < .01) after adjustment for confounders. DISCUSSION/SIGNIFICANCE OF IMPACT: In cART-era data, HIV+ NSCLC patients had a poorer prognosis than those without HIV after accounting for competing risks of death. Further research is needed to evaluate the effect of HIV specific factors, such as immunosuppression and anti-retroviral use, on lung cancer prognosis.
RACE AND CONGESTIVE HEART FAILURE: EQUAL TREATMENT, UNEQUAL OUTCOMES
Oderinde AE1, Rust G1, Zhang S1, Ofili E1, Onwuanyi A1, Lapu R1 1Morehouse School of Medicine, Atlanta, GA, USA
OBJECTIVES/SPECIFIC AIMS: We undertook this study to determine if racial disparities exist in guideline-concordant treatment and outcomes among low-income CHF patients enrolled in Medicaid; and to examine the extent to which such racial disparities if any, mediate racial disparities in clinical adverse event outcomes. METHODS/STUDY POPULATION: Retrospective cohort analysis of claims data for 10,177 Medicaid beneficiaries aged 40–64 years in fourteen southern states with CHF (2005–2007). We examined demographic characteristics, comorbidity status, and receipt of CHF-related medications. Outcomes of interest were CHF-related 90-day readmission, emergency department visits and total inpatient bed days. RESULTS/ANTICIPATED RESULTS: African American (AA) patients were actually more likely than whites to have received a guideline concordant treatment (80% versus 74%). AA were more likely than whites to receive: ACE-I or ARB (58% and 51% respectively); beta blocker (63% and 59% respectively); and both beta blocker and ACE-I or ARB (42% and 36% respectively). Although AA were more likely to receive guideline concordant treatment, their health outcomes were not better. Compared to whites, AA had significantly higher (p < 0.01) readmission rates (17.9% vs. 13.2%), total inpatient bed days (6.5 vs. 6.2), and number of emergency department visits (1.6 vs. 1.3) during the 90 days following their initial inpatient admission. In multivariate models, comorbidity status was the most significant predictor of CHF outcomes. DISCUSSION/SIGNIFICANCE OF IMPACT: Racial disparities in CHF outcomes persist even among equally-insured low-income Medicaid enrollees, despite elimination of disparities in guideline-concordant therapy for CHF. Further research is needed to determine the optimal management of clinically complex CHF patients with multimorbidity and to eliminate racial disparities in their clinical outcomes.
ROBOTIC ENDOSCOPIC ATRAUMATIC CORONARY ARTERY BYPASS GRAFTING DECREASE 30 DAYS COMPLICATION RATE, LENGTH OF STAY AND ACUTE CARE FACILITY DISCHARGE RATE COMPARED TO CONVENTIONAL CORONARY ARTERY BYPASS GRAFTING
Leyvi G1, DeRose JJ2 1Montefiore MC / AECOM, Bronx, NY, USA; 2Montefiore Medical Center /AECOM, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: It has been hypothesized that robotic endoscopic atraumatic coronary artery bypass grafting (endo-ACAB) approach can confer the survival advantages of conventional CABG while decreasing the complication rate and improving rapid recovery. METHODS/STUDY POPULATION: We retrospectively analyzed 2088 consecutive patients who underwent CABG from 2007 to March 2012. In order to compare the incidence of rapid recovery between conventional CABG and robotic endo-ACAB, the surrogate variables of early discharge (LOS < 6 days) and discharge to home (vs. rehab or acute care facility) were evaluated. A multivariate logistic regression analysis was utilized and included the variables of robotic endo-ACAB, NY State Postoperative Complication Composite, and Predicted Perioperative Morbidity and Mortality (PROMM) score in 4 quartiles. A comparison of 30 day complication rate was also performed. RESULTS/ANTICIPATED RESULTS: 150 robotic endo-ACAB cases and 1,938 conventional CABG cases were analyzed. Mutlivariate logistic regression analysis demonstrated that robotic endo-ACAB was a strong predictor of lower 30 day complications (OR = 0.3, p = 0.026), short LOS (OR 2.5, p < 0.001), and decreased need for an acute care facility (OR 0.6, p = 0.046) (table 1) adjusted for Predicted Perioperative Morbidity and Mortality (PROMM) score in four quartiles. In the presence of complications robotic endo-ACAB was not associated with a change in discharge status or short LOS. DISCUSSION/SIGNIFICANCE OF IMPACT: Robotic endo-ACAB is associated with a lower 30 day complication rate than conventional CABG. A higher incidence of short LOS and a lower incidence of acute care facility discharge may suggest a more rapid recovery to pre-operative status in these patients.
SLEEP QUALITY OF MECHANICALLY VENTILATED CHILDREN IN THE PEDIATRIC INTENSIVE CARE UNIT
Kudchadkar SR1, Yaster M1, Punjabi NM2 1Johns Hopkins, Baltimore, MD, USA; 2Johns Hopkins, Pulmonary and Critical Care Medicine, Baltimore, MD, USA
OBJECTIVES/SPECIFIC AIMS: The overall objective of this proposal is to (SA1) characterize the sleep structure of mechanically ventilated children in the PICU receiving sedative medications with polysomnography (PSG) and actigraphy, (SA2) evaluate the effect of sedation management on the sleep EEG and (SA3) investigate the effect of a sleep behavioral protocol on length of mechanical ventilation, sedative use, and hospital length of stay. METHODS/STUDY POPULATION: A prospective observational cohort design is being used to evaluate children requiring mechanical ventilation and intravenous sedative/analgesic medications in a pediatric ICU. The primary outcome variables used are total sleep time (TST) and percent-time spent in restorative sleep (REM or SWS) within the cohort compared with age-matched normative PSG data. RESULTS/ANTICIPATED RESULTS: Previously healthy children requiring mechanical ventilation and receiving opioid and/or sedative medications have a severe reduction of REM sleep (1.2% of TST vs. 20% in normal age matched controls) and significantly increased sleep fragmentation (>3 arousals/hour vs. 1–2 arousals/8 hour sleep period). Opioid and benzodiazepine dose/day correlated with an increase in sleep fragmentation. We anticipate that a sleep behavioral protocol using noise reduction and light optimization will result in improved sleep quality, decreased sedative needs and shortened hospital length of stay. DISCUSSION/SIGNIFICANCE OF IMPACT: Mechanically ventilated children suffer a severe disruption of restorative sleep. Further data will provide us the first hypothesis driven evaluation of sleep in critically ill children and a quantifiable metric that we will use in developing strategies to improve sleep and sedation management in mechanically ventilated children in the future.
SOCIAL DETERMINATS OF ORAL HEALTH IN PREGNANT WOMEN IN PUERTO RICO
Medina-Paneto JA1, López L1, Elias A1, Russell S2 1University of Puerto Rico-Medical Sciences Campus, San Juan, USA; 2New York University, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: It has been established that poor oral hygiene is the precursor of poor oral health. Poor oral health includes periodontal diseases. Although is actually controversial, periodontal disease has been related to complications in pregnancy and preterm birth. Also, low-income families and ethnic minorities' women have a higher risk of low-birth weight babies. Access to dental care utilization is low among pregnant women, even though they are often eligible for higher levels of dental insurance coverage. There is limited evidence in the relationship between oral health and complications during pregnancy and childbirth in Puerto Rico, even though that 20% of all live births are premature. METHODS/STUDY POPULATION: A correlation study will be performed to assess the relationship of social determinants of health, such as, socioeconomic status, educational level and geographical residence location, and the knowledge of oral health status during pregnancy in Puerto Rican women. The sample will be obtained from the OB/Gyn Clinic at the UPR-Hospital in Carolina, PR and the Intramural OB/Gyn Clinic of the UPR-School of Medicine. The information will be obtained from a questionnaire designed to collect the knowledge about oral health during pregnancy and demographic information, such as level of education, location of residence, health care insurance and age. RESULTS/ANTICIPATED RESULTS: We expect to identify which social determinants are involved in the adequate access to oral health care during pregnancy. DISCUSSION/SIGNIFICANCE OF IMPACT: Therefore, be able to develop pre-natal care protocols that contribute to the decrease of pregnancy complications and pre-term births in Puerto Rico. This project is partially supported by The NIH-AN R25MD007607 NIMHHD.
SURVIVAL FOLLOWING A RANDOMIZED TRIAL OF LIGHT VS DEEP SEDATION IN PATIENTS UNDERGOING SURGICAL REPAIR OF A HIP FRACTURE UNDER SPINAL ANESTHESIA
Brown C1, Gottschalk A1, Sieber F1 1Johns Hopkins, Baltimore, MD, USA
OBJECTIVES/SPECIFIC AIMS: Low intraoperative Bispectral Index (BIS) values (range 0–100) have been associated with decreased survival. Previously, we randomized 114 patients undergoing hip fracture surgery to deep sedation (BIS∼50) or light sedation (BIS > 80). The aim of this study was to determine whether light sedation would improve 1-year survival in the patients from this trial. METHODS/STUDY POPULATION: Patients older than 65 with a traumatic hip fracture were eligible to be enrolled in a randomized controlled trial, which assessed whether decreasing the depth of intraoperative levels of sedation during a spinal anesthetic could prevent delirium. Patients were randomized to deep sedation (BIS∼50) or light sedation (BIS > 80). The primary outcome of this study was survival at 1 year post-surgery. Kaplan-Meier curves were calculated based on sedation level, and Cox proportional hazard models were used to compare survival functions. Significant co-morbidities were those associated with overall mortality in univariable analysis. RESULTS/ANTICIPATED RESULTS: At one-year follow-up among all patients, we found no statistically significant difference in survival in the light sedation group compared to the deep sedation group (HR 0.59, 95% CI 0.28–1.26, P = 0.18). However, among patients with at least one severe co-morbidity, we found a significant reduction in mortality in the light sedation group compared to the deep sedation group (HR 0.42, 95% CI 0.18–0.98, P-0.046). DISCUSSION/SIGNIFICANCE OF IMPACT: Among patients with at least one significant co-morbidity undergoing surgical repair of a hip fracture under spinal anesthesia, light sedation resulted in improved one-year survival.
THE FILIPINO FAMILY INITIATIVE: PRELIMINARY EFFECTS OF AN EVIDENCE-BASED PARENTING PROGRAM OFFERED IN CHURCHES ON PARENT OUTCOMES
Javier JR1, Coffey DM1, Schrager SM1, Palinkas LA1 1Southern California Clinical and Translational Science Institute, Los Angeles, CA, USA
OBJECTIVES/SPECIFIC AIMS: Filipinos, the second largest U.S. immigrant population, have significant disparities in youth behavioral health outcomes compared to non-Hispanic whites. Parenting programs provided in childhood are effective in preventing the onset and escalation of behavioral health problems. The purpose of this study was to pilot-test an evidence-based parenting program offered in churches as a behavioral health prevention intervention for Filipino youth. METHODS/STUDY POPULATION: Filipino parents of children ages 6–12 years participated in a 12-week parenting intervention using the Incredible Years (IY) Program. Parents completed measures of parenting practices (positive verbal discipline and use of physical punishment) and parenting-related stress before and after the intervention, and satisfaction was assessed after the program. Paired t-tests compared mean parenting and stress scores; satisfaction was assessed descriptively. RESULTS/ANTICIPATED RESULTS: 22/28 (78%) parents completed pre- and post- intervention assessments and the 12-week IY program. Parents reported significant improvements in positive verbal discipline (mean (SD) difference = 0.46(0.52), p < 0.05), a marginal decrease in physical punishment (mean (SD) difference = –0.25(0.66), p < .10), and significant reductions in stress (mean (SD) difference = –9.23(20.26), p < 0.05) following the IY program. Families reported very high satisfaction with the content and format of IY (means ranged from 5.73 to 6.95 out of 7). 81% of parents wanted to keep meeting as a group after the intervention ended. DISCUSSION/SIGNIFICANCE OF IMPACT: Results support the benefits and feasibility of providing an evidence-based parenting program to Filipino parents of school-age children in faith-based settings in order to prevent future behavioral health problems.
THE IMPACT OF SOCIOECONOMIC STATUS ON SURVIVAL IN CYSTIC FIBROSIS LUNG TRANSPLANTATION
Braun AT1, Merlo CA1 1Johns Hopkins University, Baltimore, MD, USA
OBJECTIVES/SPECIFIC AIMS: To evaluate the impact of educational attainment and health insurance status on survival in cystic fibrosis (CF) lung transplant (LTx) recipients. METHODS/STUDY POPULATION: Retrospective review of United Network of Organ Sharing (UNOS) data for 2,085 adult CF LTx recipients from 2000 through 2010. All-cause mortality was examined with multivariable Cox proportional hazard regression, and the Kaplan-Meier method was used to model survival after lung transplantation. RESULTS/ANTICIPATED RESULTS: Of 2,085 CF LTx recipients, 1,323 (63.5%) had private insurance; 762 (36.5%) had non-private insurance. In regard to education, 132 (9.9%) patients had not completed high school, 762 (36.6%) had a high school degree, and 1,028 (53.5%) had any college. During the study period, 778 patients (37.3%) died. Patients with non-private insurance had 15% lower 5-year survival than patients with private insurance. A survival difference was also noted at 1-year (4%), 2-years (6%), and 3-years (9%). An educational attainment of high school or less was associated with a 17% lower survival at 5-years post transplantation when compared to individuals with any college education. This survival difference was also noted at 2-years (7%) and 3-years (11%), but not at 1-year. In multivariable analyses, non-private insurance status increased the risk of death (hazard ratio, 1.33; 95% confidence interval, 1.14 to 1.55). Similarly, educational attainment of high school or less was associated with an increased risk of death (hazard ratio, 1.31; 95% confidence interval 1.10 to 1.54). DISCUSSION/SIGNIFICANCE OF IMPACT: Improved survival is seen in patients with private insurance and higher levels of educational attainment. Further investigation of health care delivery mechanisms and patient-specific behaviors may aid in improving the utilization and outcomes of the limited and lifesaving resource of lung transplantation.
THERAPEUTIC DELAYS LEAD TO WORSE SURVIVAL AMONG PATIENTS WITH HEPATOCELLULAR CARCINOMA
Singal A1, Patel N1, Tiro J1, Marrero J1, Yopp A1 1University of Texas Southwestern, Dallas, TX, USA
OBJECTIVES/SPECIFIC AIMS: Although prior studies have demonstrated underutilization of appropriate therapy in patients with hepatocellular carcinoma (HCC), no studies to date have assessed the prevalence and clinical impact of therapeutic delays among HCC patients. The aim of our study was to characterize and identify factors associated with underutilization and delays in treatment among patients with HCC. METHODS/STUDY POPULATION: We conducted a retrospective cohort study of cirrhotic patients diagnosed with HCC at a large urban safety-net hospital between January 2005 and June 2012. Dates for HCC diagnosis and any treatments were recorded. Univariate and multivariate analysis was used to determine factors associated with treatment underutilization and therapeutic delays, which was defined as time from diagnosis to treatment exceeding three months. RESULTS/ANTICIPATED RESULTS: We identified 267 treatment-eligible patients with HCC, of whom only 62% received HCC therapy. On multivariate analysis, tumor stage (OR 0.48, 95%CI 0.36–0.65), Child Pugh class (OR 0.49, 95%CI 0.28–0.84), and Black race (OR 0.55, 95%CI 0.31–0.99) were associated with lower rates of treatment utilization. The median time to treatment was 1.7 months, with 31% of patients experiencing delayed treatment. Delayed treatment was associated with the presence of ascites (HR 2.8, 95%CI 1.3–6.1) and undergoing transarterial chemoembolization (HR 4.8, 95%CI 1.8–12.5). After adjusting for tumor stage and Child Pugh class, treatment underutilization (HR 0.33, 95%CI 0.24–0.46) and therapeutic delays (HR 0.50 95%CI 0.30–0.84) were both associated with significantly worse survival. DISCUSSION/SIGNIFICANCE OF IMPACT: In addition to one-third of patients not receiving HCC-directed therapy, another 20% experience significant therapeutic delays, leading to worse survival.
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- SCHOLAR/TRAINEE ABSTRACTS
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
- Research Professionals Abstracts Best Practices
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
DESCRIPTION AND PRODUCTIVITY OF EMERGENCY MEDICINE RESEARCHERS RECEIVING K23 OR K08 MENTORED RESEARCH CAREER DEVELOPMENT AWARDS
Nishijima DK1 1UC Davis School of Medicine, Sacramento, CA, USA
OBJECTIVES/SPECIFIC AIMS: The objective of this study was to describe emergency medicine (EM) researchers who have received a K08 or K23 award and to evaluate their post-award federal funding and publications. METHODS/STUDY POPULATION: We conducted a cross-sectional study during Jan 2012 of clinician scientists who previously completed an EM residency or fellowship and have received a K08 or K23 from NIH or AHRQ between fiscal years 2000–2011. Awardees were identified and post-award funding were abstracted from NIH RePORTER. Publications and impact factor were calculated from PubMed and Journal Citation Reports. We also conducted an email survey of all EM K awardees during Sept and Oct 2012 to describe awardee characteristics. Simple descriptive statistics were reported. RESULTS/ANTICIPATED RESULTS: 63 EM awardees were identified; 24 (38%) were K08 awardees and 39 (62%) were K23 awardees. Of the 38 (60%) awardees who completed their training, 16 (42%) obtained subsequent federal funding with 6 (16%) obtaining R01 funding (median time from end of K to R01 of 4.5 years). The mean number of publications per awardee year was 4.0 with a mean five-year impact factor score of 4.5 per publication. Forty-five (71%) of EM K awardees responded to the survey. Respondents had a median age of 36 (IQR 25–38) and 33 were male (73%). The median time from completion of residency to K award was 6 years (IQR 4–7 years) with 27 (60%) completing a research fellowship prior to K award. DISCUSSION/SIGNIFICANCE OF IMPACT: Approximately one in six EM K awardees who completed their K08 or K23 training subsequently obtained R01 equivalent funding. Awardees published a median of 4.0 manuscripts per year in journals with a mean impact factor of 4.5. Given the numerous barriers facing EM researchers, EM K awardees demonstrate good post-award productivity.
EDUCATING TRANSLATIONAL PHYSICIAN-SCIENTISTS: THE NYU SCHOOL OF MEDICINE MD/MS DUAL DEGREE IN TRANSLATIONAL INVESTIGATION
Maddalo S1, Fierman A1, Schwartz M1, Pillinger M1, Gold-Von Simson G1 1New York University School of Medicine CTSI, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: There is a growing need for interdisciplinary, multi-institutional, international collaboration in medical research. The NIH encourages educational institutions to provide students with training to integrate vital knowledge of bench, bedside, and community research. The NYUSOM Masters of Science in Clinical Investigation originated in 2005 as a post-doctoral NIH K-30 grant-supported program designed to impart these skills and has proven to be immensely successful as measured by grant generation and healthcare impact. As a result the CTSI, in concert with the SOM, created a dual-degree track for medical students within the overarching program. METHODS/STUDY POPULATION: Medical students are chosen by a competitive application process. Applicants must have identified a well-established research mentor and clinically relevant translational project. A rigorous curriculum in the first year is well developed and includes research design, biostatistics, and drug development among other core courses. In the second year, students focus on their mentored-research and meet with MSCI directors/meta-mentors each semester for dynamic interface. RESULTS/ANTICIPATED RESULTS: Program requirements and research benchmarks are assessed on a semester basis. A research thesis is defended at the end of year 2. To date, 9 abstracts and 2 papers have been published by medical students in this program. An additional 5 abstracts and 6 papers have been submitted by students. DISCUSSION/SIGNIFICANCE OF IMPACT: As has been the case with the MSCI program, medical students will develop essential skills that will enable them to engage in the meaningful translational research. The dual degree program at the NYUSOM will provide the foundation for these future physician researchers to design, develop, and implement projects that will improve patient care.
STANFORD UNIVERSITY NETWORK FOR DIAGNOSIS OF RETINOPATHY OF PREMATURITY (SUNDROP): SEVEN YEARS OF SCREENING
Fijalkowski N1, Wang SK1, Henderson MT1, Leng T1, Moshfeghi AA2, Moshfeghi DM1 1Stanford School of Medicine Byers Eye Institute, Palo Alto, CA, USA; 2University of Miami School of Medicine Bascom Palmer Eye Institute, Miami, FL, USA
OBJECTIVES/SPECIFIC AIMS: To report the seven-year experience of the SUNDROP telemedicine initiative, which was developed to reduce the risk of blindness from retinopathy of prematurity (ROP). METHODS/STUDY POPULATION: A retrospective analysis of the SUNDROP archival data. Nurses screened 608 infants at four community NICUs with RetCam II. An ROP specialist evaluated these images remotely at Stanford. Every infant then received a bedside examination within one week of discharge. All infants were screened with telemedicine according to recommended timelines. Primary outcomes were treatment-warranted ROP (TW-ROP) and anatomical outcomes. RESULTS/ANTICIPATED RESULTS: The SUNDROP telemedicine screening initiative did not miss any TW-ROP at seven-years. A total of 608 infants were imaged in 2,169 exams. 22 infants had TW-ROP on image review, 21 of these were TW-ROP after bedside exam. Infants with TW-ROP had lower gestational age (24.8 wks), lower birth weight (819 g), and were more likely male than the no TW-ROP cohort (p-values < 0.00001). The sensitivity of telemedicine's ability to detect TW-ROP was 100%, specificity of 99.8%, positive predicative value of 95.5% and negative predictive value of 100%. No patient progressed to retinal detachment or any adverse anatomical outcome. DISCUSSION/SIGNIFICANCE OF IMPACT: The SUNDROP telemedicine initiative demonstrated a high degree of diagnostic reliability and was able to capture all infants with TW-ROP. Telemedicine offers a reliable, cost-effective and accurate method of identifying TW-ROP without sacrificing quality of care or outcomes.
- Top of page
- SCHOLAR/TRAINEE ABSTRACTS
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
- Research Professionals Abstracts Best Practices
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
DETAILED LONGITUDINAL SAMPLING OF THE WOUND MICROBIOME REVEALS INCREASED BACTERIAL DIVERSITY WITH HEALING
Tuttle MS1, Sprockett DD1, Ghannoum MA1 1Case Western Reserve University, Cleveland, OH, USA
OBJECTIVES/SPECIFIC AIMS: A striking clinical feature of chronic wounds is their high level of microbial bio-burden. However, prior culture-based studies on the association between specific microbes and wound healing have been inconclusive. We used culture-independent methods to prospectively characterize changes in wound bacterial communities in a patient with a chronic venous leg ulcer. METHODS/STUDY POPULATION: The patient was sampled bi-weekly over a healing course of 26 weeks while receiving standard therapy including debridement, compression bandages, and wound dressings containing topical silver as deemed necessary by the clinician. RESULTS/ANTICIPATED RESULTS: Next generation sequencing of the 16S ribosomal RNA bacterial gene revealed high concordance in bacterial community structure regardless of location sampled within the wound (distal, central, and proximal wound bed). However, superficial bacterial communities were distinct from deeper tissue samples, with the former being less diverse than the latter (Shannon diversity 2.07 vs. 2.76, p = 0.02). These results suggest that consistent sampling depth is more critical than sample location within the wound. Notably, we found that deep wound tissues experienced two distinct shifts in bacterial community structure: from a community dominated by Acinetobacter in the first 6 weeks (average 60.96%) to a community dominated by Staphylococcus until healing (average 48.03%), as well as a marked increase in bacterial diversity over the final 8 weeks of healing (Shannon diversity 3.86 vs. 2.23, p = 0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: These results suggest that wound microbial dynamics shift throughout the course of healing, with a trend toward increased taxonomic diversity with healing, underscoring the need for further longitudinal study designs when determining the microbial influence on healing outcomes in chronic wounds.
A PILOT STUDY OF EVALUATION OF A PHOSPHORYLATION SIGNATURE FROM PANCREATIC CANCERS
Wang-Gillam A1, Langley E2, Philip K2, Gao F1, Azar R1 1Washington University in St. Louis, St. Louis, MO, USA; 2Prometheus, San Diego, CA, USA
OBJECTIVES/SPECIFIC AIMS: The development of targeted therapy in pancreatic cancer has been hindered by minimal material available for molecular analysis from diagnostic endoscopic ultrasound-guided fine-needle aspirations (EUS/FNA). Revealing the phosphorylation status of key pathway mediators would allow optimal selection of patients for kinase inhibitor clinical trials. We launched a pilot study to detect phosphorylation signatures from FNA samples using an ultrasensitive multiplexed protein microarray platform (CEER, Prometheus). METHODS/STUDY POPULATION: Patients who underwent diagnostic EUS/FNA for a suspicious lesion in the pancreas underwent two additional EUS passes. Mediators of multiple signaling pathways including HER1/EGFR, HER2, HER3, IGF-IR, and their downstream mediators, AKT, MEK, ERK, et al were evaluated by CEER. RESULTS/ANTICIPATED RESULTS: The study has 57 patients enrolled with a planned final enrollment of 100. This is an interim analysis. Thirty-nine (68.4%) had carcinoma cytology, 9 had atypical and 9 had negative cytology. HER2 (median value of 18 CU vs. 2 CU, P = 0.021) and HER3 (median value of 547 CU vs. 168 CU, P = 0.008) showed significantly higher levels of phosphorylation in malignant FNAs over their counterparts with negative cytology. Robust levels of CK expression were also observed in FNAs with carcinoma cytology (median CK level 1030 CU vs. 106 CU, P = 0.007). The wide range of phosphorylation signatures detected indicates this is a feasible clinical approach. DISCUSSION/SIGNIFICANCE OF IMPACT: Profiling the phosphorylation status of pancreatic cancer pathways from FNA samples is feasible. If further validated, patients with pancreatic cancer can be screened for targeted therapy by their phosphorylation signature.
A POTENTIAL BIOMARKER FOR FOCAL DYSTONIA: ABNORMAL DORSAL PREMOTOR-MOTOR INHIBITION IN CERVICAL DYSTONIA
Pirio Richardson S1, Radigan M1 1University of New Mexico, Albuquerque, NM, USA
OBJECTIVES/SPECIFIC AIMS: In cervical dystonia (CD), dysfunctional synaptic regulation between dorsal premotor and motor cortex could explain the abnormal motor control seen in these patients when trying to maintain normal head posture. Previous work has shown an abnormal increase in dorsal premotor-motor intracortical inhibition (dPMI) in patients with focal hand dystonia. This project aims to determine whether there is enhanced dPMI in CD patients. METHODS/STUDY POPULATION: We have studied four control subjects and four patients with CD. dPMI was tested by applying conditioning transcranial magnetic stimulation to the left dorsal premotor cortex and then a test pulse to the ipsilateral motor cortex at an interval of 6 ms. dPMI was tested at rest and with contraction of the first dorsal interosseousmuscle at 10% maximum force. We also looked at the duration of the cortical silent period (CSP)—a measure of cortical excitability—during the contraction task. RESULTS/ANTICIPATED RESULTS: Our results show that patients with CD have enhanced dPMI at rest (86.3%, the ratio of conditioned to unconditioned test pulse) in contrast to controls (130.6%). The difference between patients and controls persisted with contraction (89.9% vs. 115.2%). Patients with CD showed a shorter CSP than controls (116.7 ms vs. 189.0 ms). DISCUSSION/SIGNIFICANCE OF IMPACT: These preliminary results suggest that abnormal premotor-motor interaction may indeed be a biomarker for the identification of patients with focal dystonia. Given that the CSP is shortened in dystonia patients, this suggests an enhanced cortical excitability consistent with loss of inhibition in the pathophysiology of dystonia. In this setting, the enhanced dPMI may be a compensatory response. Successful completion of this study could lead to enhanced diagnosis of focal dystonia as well as providing a potential target for therapeutic intervention.
A POTENTIAL ROLE FOR OREXIN IN REGULATING CIRCADIAN FLUCTUATION OF INTRAOCULAR PRESSURE
Samuels BC1, Hammes NM1, Renger JJ2, Gotter AL2, Winrow CJ2, Johnson PL1, Shekhar A1 1Eugene & Marilyn Glick Eye Inst, Ind Univ, Indianapolis, IN, USA; 2Merck Research Laboratories, West Point, PA, USA
OBJECTIVES/SPECIFIC AIMS: Circadian fluctuation in intraocular pressure (IOP) has recently been identified as an independent risk factor for glaucoma progression. Neurons in the dorsomedial and perifornical hypothalamus (DMH/PeF) are known to regulate circadian rhythms, and we have recently shown that chemical stimulation of this region evokes increases in IOP. Orexins are a novel class of neuropeptides located almost exclusively in the DMH/PeF region and are known to play a key role in regulating circadian behaviors and autonomic functions. We hypothesize that fluctuations in IOP is regulated, at least in part, by these orexin containing neurons. METHODS/STUDY POPULATION: A dual orexin-1 and orexin-2 receptor antagonist (DORA; 30 mg/kg or 10 mg/kg) or vehicle control was systemically administered to isoflurane-anesthetized rats 75-min. prior to chemical stimulation of the DMH/PeF region by stereotaxically guided microinjection of a GABAA receptor antagonist bicuculline methoidide (30 pmol/75 nL). Intracranial pressure (ICP) and cardiovascular activity were continuously monitored. An iCareLab tonometer was used to record IOP every 2 min. RESULTS/ANTICIPATED RESULTS: Chemical stimulation of the DMH/PeF evoked significant increases in IOP that followed a cardioexcitatory response and increase in ICP, similar to our previous results. Prior administration of a DORA attenuated the increases in IOP in a dose dependent fashion. DISCUSSION/SIGNIFICANCE OF IMPACT: These data are the first to support the hypothesis that orexin neurons located in the DMH/PeF region play a role in mediating circadian fluctuations in IOP, a previously unrecognized function of these neurons. Further, the orexin system may provide a novel target for future glaucoma therapies aimed at reducing circadian fluctuation of IOP.
A ROLE FOR PAK-1 MEDIATED SIGNALING IN THE PREVENTION OF BETA-CELL FAILURE
Wang Z1, DiMeglio LA1, Mather K1, Thurmond D1 1Indiana University School of Medicine, Indianapolis, IN, USA
OBJECTIVES/SPECIFIC AIMS: Type 2 diabetes is ultimately a result of decreased pancreatic β-cell mass. Agents that prevent the progressive decline in β-cell mass offer the potential to slow or possibly reverse diabetes development. The p21-activated kinase 1 (Pak1) is a serine/threonine kinase that has multiple functions including cell survival; however its role in β-cell survival and replication has never been addressed. METHODS/STUDY POPULATION: Beta-cell survival and proliferation were examined using pancreatic MIN6 β-cells, Pak1+/- heterozygous mouse islets and human islets. RESULTS/ANTICIPATED RESULTS: We observed that Pak1 is critical for β-cell survival and proliferation. Pak1-depletion induced significant cell death in MIN6 cells. Decreased cell proliferation was observed in Pak1+/- mouse islets and Pak1-depleted MIN6 cells as determined by incorporation rate of [3H]methyl-thymidine into genomic DNA. We also identified that Pak1 beneficially regulates BAD and Survivin, critical proteins in β-cell apoptosis and replication, respectively. Exogenous PAK1 prevented decreased Survivin and increased BAD concentrations in MIN6 cells subjected to hyperglycemic stress. In effort to screen potential Pak1 activators from commonly-used supplements, we observed that Ginsenoside Rb1(Gs-Rb1), the main ingredient in Ginseng, significantly increased Pak1 expression and phosphorylation/activation, concurrent with increased Survivin and decreased BAD protein abundances. Gs-Rb1 also significantly counteracted the effects of high glucose to up-regulate BAD and induce apoptosis in cultured human islets. DISCUSSION/SIGNIFICANCE OF IMPACT: Our data implicate Pak1 as a potential molecular target for the restoration of β-cell mass. Moreover, commonly-used dietary supplement Ginseng may mediate β-cell survival and replication via Pak1 signaling.
A TRANSLATION MODEL OF AUTOIMMUNE PSYCHIATRIC ILLESS
Williams K1 1Yale Child Study Center, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Interest and controversy have surrounded a possible autoimmune etiology for childhood obsessive compulsive (OC) symptoms and tic disorders (TD). Termed PANDAS (Pediatric Autoimmune Neuropsychiatric Disorder Associated with Streptococcal infections), PANDAS is characterized by a rapid onset of OC and tic symptomatology following a Group A Streptococcal (GAS) infection. Evidence suggests that treatment with anti-autoimmune therapies, such as Intravenous Immunoglobulin (IVIG) rapidly improves OC and tic symptoms in PANDAS children. Evidence also suggests that intracerebral infusion of serum from patients with tic disorders into the striatum of rats is capable of acute induction of motor stereotypies. The current study seeks to develop a translational model of PANDAS utilizing serum collected from patients enrolled in a clinical trial of IVIG in the treatment of PANDAS, and infusing it into mice. METHODS/STUDY POPULATION: Patients who meet the criteria for PANDAS are currently enrolled in a study of IVIG therapy conducted by Yale and the NIMH. Mice will be injected intracerebrally with serum taken from individuals in the PANDAS IVIG trial. Four experimental groups will be used;Mice will be injected with 1) pre-IVIG treatment patient serum, 2) post-IVIG treatment serum, 3) non-PANDAS childhood onset OC/tic, and 4) normal human serum. Tests of anxiety and compulsive grooming will be assessed before and after serum injection. Behavioral testing will be conducted using automated behavioral recording systems, with experimenters blinded to the type of serum infused. Group differences in the presence of behavioral and motor abnormalities will be assessed using repeated-measures analyses. RESULTS/ANTICIPATED RESULTS: These experiments are ongoing. DISCUSSION/SIGNIFICANCE OF IMPACT: If successful, these experiments will provide a translational model of autoimmune psychiatric illness, and further our mechanistic understanding of PANDAS symptomatology.
ADIPONECTIN DECREASES THE SURVIVAL OF HUMAN MULTIPLE MYELOMA CELL LINES: PKA/AMPK-MEDIATED INDUCTION OF APOPTOSIS
Medina EA1, Oberheu K1 1University of Texas Health Science Center San Antonio, San Antonio, TX, USA
OBJECTIVES/SPECIFIC AIMS: MM is incurable and characterized by the accumulation of neoplastic plasma cells in the bone marrow. Epidemiologic studies suggest that obesity increases the risk of MM. Adiponectin is produced primarily by adipocytes but paradoxically decreased in obesity. The adipokine has been demonstrated to inhibit the proliferation of some epithelial cancer cell lines. We explored the effects of adiponectin on the survival of human MM cell lines (RPMI8226, U266, MM1S and MM1R). METHODS/STUDY POPULATION: n/a RESULTS/ANTICIPATED RESULTS: The MM cell lines expressed adiponectin receptors AdipoR1 and AdipoR2. Adiponectin decreased the survival of MM cells in a concentration- and time-dependent manner via inducing apoptosis. Adiponectin activated AMPK and inhibited basal levels of AKT activation. MM cells exposed to AICAR or metformin (well-known AMPK activators) activated AMPK, decreased basal AKT activation and induced apoptosis. Inhibiting PKA with H89 (PKA inhibitor) suppressed adiponectin-induced AMPK activation, inhibition of basal AKT activation and apoptosis. Forskolin, which increases intracellular cAMP levels and thereby activates PKA, mimicked the effects of adiponectin; pretreatment with H89 dampened forskolin's effects. In contrast, H89 did not suppress AICAR or metformin-induced AMPK activation, inhibition of basal AKT activation or apoptosis. DISCUSSION/SIGNIFICANCE OF IMPACT: Our data suggest that apoptosis of human MM cells by adiponectin is mediated via activation of PKA, which leads to AMPK activation and inhibition of basal AKT activation. Adiponectin, AICAR, metformin and other agents that activate PKA and/or AMPK may be utilized to treat MM. Moreover, decreased circulating adiponectin levels, as occurs in obesity, may promote MM growth. Future studies will target the PKA/AMPK pathway in a pre-clinical syngeneic mouse MM model.
ADMA LEVELS POST KIDNEY TRANSPLANT
Birdwell K1, Graves AJ1, Laderbush BF1, Shintani A1, Ikizler TA1 1Vanderbilt University, Nashville, TN, USA
OBJECTIVES/SPECIFIC AIMS: Asymmetrical dimethylarginine (ADMA) is positively associated with cardiovascular disease in the general population. Chronic kidney disease is associated with elevated ADMA, for a variety of proposed factors including decreased glomerular function (GFR) and enhanced vascular dysfunction. We hypothesized that ADMA levels would decrease over time post kidney transplant while accounting for change in kidney function, indicating improved vascular function in the transplanted state. METHODS/STUDY POPULATION: In a prospective cohort of 91 kidney transplant recipients, we collected medical information and plasma ADMA levels pre transplant and 1 week, 1 month, and 6 months post transplant. We used a linear mixed effect model to assess the association between ADMA and time, with and without adjustment for creatinine, and to assess the association between creatinine and time. A linear mixed effect model was also used to assess whether any factors were associated with change in ADMA from baseline to post-transplant times. Clinical covariates were selected a priori, including sex, race, diabetes, cardiovascular disease, and smoking status. RESULTS/ANTICIPATED RESULTS: The cohort was 30.8% female and 19.8% African-American. Median ADMA and creatinine levels prior to transplant were 0.41 micromolar and 6.9 mg/dL, respectively. After transplant, ADMA levels significantly decreased between baseline and post-transplant times (p = 0.049). Serum creatinine also decreased between baseline and post-transplant (p < 0.0001). There was no significant decrease in ADMA over time after adjusting for creatinine (p = 0.62). No covariates were associated with change in ADMA from baseline (all p > 0.08). DISCUSSION/SIGNIFICANCE OF IMPACT: ADMA and creatinine levels changed in the first 6 months after transplant, though no factors were associated with change in ADMA, suggesting further study to understand why ADMA levels decrease.
ADVERSE CHILDHOOD EXPERIENCES (ACE) PREDICT HEAVIER DRINKING AND GREATER ALCOHOL INTAKE DURING INTRAVENOUS (IV) ALCOHOL SELF-ADMINISTRATION IN NON-DEPENDENT DRINKERS
Stangl BL1, Schwandt ML1, Ramchandani VA1 1NIAAA, Bethesda, MD, USA
OBJECTIVES/SPECIFIC AIMS: ACEs have been associated with alcohol use disorders; however the relationship between ACEs and alcohol intake in non-dependent drinkers is less clear. This study examined the influence of ACE on drinking history and IV alcohol self-administration, using the Computer-Assisted Self-infusion of Ethanol (CASE) method that allows individuals to self-administer IV alcohol. METHODS/STUDY POPULATION: Non-dependent drinkers (N = 201) were assessed for ACE using the Childhood Trauma Questionnaire (CTQ) and for recent drinking history using the Timeline Follow Back (TLFB). Alcohol intake behavior was measured in a subset (N = 51) during a CASE session, consisting of a 25-min priming phase of 4 standardized alcohol infusions, followed by a 125-min open-bar phase where they could push a button for additional infusions. Self-administration measures included peak (PEAK) and average (AVG) BrAC and total ethanol (EtOH). RESULTS/ANTICIPATED RESULTS: At least 39% reported one form of childhood trauma. TLFB measures were linearly related to the CTQ total score (all p < 0.001) as well as the number of CTQ categories (p < 0.02). Those that reported childhood emotional abuse, emotional and physical neglect had heavier drinking histories (p < 0.05). The same relationships were seen in the CASE subset. The number of CTQ categories was associated with EtOH and AVG (p < 0.05), and showed trends for association with other intake measures. TLFB measures were associated with AVG, PEAK, and EtOH (p < 0.01) in this group. DISCUSSION/SIGNIFICANCE OF IMPACT: Childhood trauma showed an effect on recent drinking history measures in non-dependent drinkers, as well as on the amount of ethanol self-administered during the CASE session. These results imply that ACEs can impact alcohol drinking in non-dependent drinkers.
AKT AND AR SIGNALING COOPERATE TO MEDIATE CELL SURVIVAL IN ADVANCED PROSTATE CANCER
She Q1, Ye Q1, Cai W1 1University of Kentucky, Lexington, KY, USA
OBJECTIVES/SPECIFIC AIMS: The PI3K/AKT and androgen receptor (AR) signaling pathways are often concurrently activated and cooperate to promote prostate cancer progression. The objective of this study was to determine the mechanisms of AKT and AR cooperation in prostate cancer growth and whether inhibition of both AKT and AR signaling is an effective approach to therapy. METHODS/STUDY POPULATION: Selective inhibitors of AKT and AR, a variety of molecular techniques, tissue culture and xenograft tumor models were used to determine the effects of AKT and AR inhibition, alone and in combination, on AKT or AR pathway activity, cell proliferation and survival, and tumor growth in advanced prostate cancer models. RESULTS/ANTICIPATED RESULTS: We showed that prostate tumors with mutational activation of AKT signaling were sensitive to AKT inhibition. In contrast, tumors with coexistent activation of the AR signaling pathway were resistant to AKT inhibition. In such tumors, inhibition of either AKT or AR alone was insufficient to induce apoptosis and to inhibit tumor growth. However, combined inhibition of both AKT and AR synergistically induced apoptosis and effectively repressed the growth of prostate cancer xenografts including the model that is resistant to castration. Notably, we identified that AKT and AR signaling cooperated to regulate expression of the anti-apoptotic protein survivin. Reduction of survivin expression by using a clinical-grade survivin suppressant YM155 markedly caused prostate cancer cell death. DISCUSSION/SIGNIFICANCE OF IMPACT: These data demonstrate that both AKT and AR signaling pathways are required for prostate cancer cell survival and cooperate to promote tumor growth by convergent regulation of survivin expression. Therefore, inhibition of both pathways or direct inhibition of survivin expression may represent an effective therapeutic strategy for advanced prostate cancers including tumors with resistance to castration.
ALTERED HOMEOSTASIS OF CD8+ T LYMPHOCYTES IN PATIENTS PRESENTING WITH ACUTE CORONARY SYNDROMES
Zidar DA1, Sparks S2, Funderburg NT3, Lederman MM3, Weinhold KJ2 1Harrington Heart and Vascular Center; Case Western Reserve School of Medicine, Cleveland, OH, USA; 2Department of Surgery, Duke University Medical Center, Durham, NC, USA; 3Division of Infectious Diseases, Department of Medicine, Case Western School of Medicine, Cleveland, OH, USA
OBJECTIVES/SPECIFIC AIMS: Inflammation plays a key role in acute coronary syndromes (ACS), but the pattern of immune activation is incompletely characterized. We recently described a pattern of monocyte activation that correlates with ACS. Here, we sought to determine whether ACS is also accompanied by altered proportions of T lymphocyte subsets. METHODS/STUDY POPULATION: We performed a case-control pilot study of 24 patients using a nine-color flow cytometry panel to simultaneously analyze T lymphocyte lineage, maturation, and cytokine production. An additional 87 patients were studied as an independent replication cohort. RESULTS/ANTICIPATED RESULTS: Fourteen patients with ACS (cases) and ten patients with stable symptoms (controls) were analyzed in the initial experimental cohort. ACS was associated with a decrease in circulating naïve CD8+ T lymphocytes and an increase in effector and memory CD8+ T lymphocytes. The cytokine production profile of phenotypically naïve CD8+ T cells was altered in ACS patients such that these cells produced more interferon (IFN)-γ and less interleukin (IL)-2. In a replication cohort, the proportion of naïve CD8+ T cells was again diminished in patients presenting with ACS and did not depend on the burden of coronary artery disease as assessed by coronary artery calcium scoring. DISCUSSION/SIGNIFICANCE OF IMPACT: ACS is associated with altered homeostasis of circulating CD8+ T lymphocytes. Additional study is needed to determine the operant mechanisms, whether these cells contribute to the pathophysiology of atherosclerosis, and if the quantification of immune cell phenotypes can serve as a novel ACS biomarker.
ANALYSIS OF CARDIAC FUNCTION IN A PEDIATRIC MOUSE MODEL OF DOXORUBICIN-INDUCED CARDIOTOXICITY USING ECHOCARDIOGRAPHIC STRAIN IMAGING
Aune GJ1,2, Andrews T1,2, Lindsey ML1,3 1University of Texas Health Science Center San Antonio, San Antonio, TX, USA; 2Greehey Children's Cancer Research Institute, San Antonio, TX, USA; 3Barshop Institute for Longevity and Aging, San Antonio, TX, USA
OBJECTIVES/SPECIFIC AIMS: Here, we report the feasibility and reproducibility of using echocardiographic longitudinal strain imaging in a pediatric mouse model of anthracycline-induced cardiotoxicity. METHODS/STUDY POPULATION: C57/BL6J mice (2 weeks old) were injected intraperitoneally with saline as a control or three different doses of doxorubicin (1 mg/kg, 3 mg/kg, and 5 mg/kg) weekly for a total of three weeks. Serial echocardiographic analysis using the VisualSonics Vevo 2100 ultrasound was performed at 3 and 5 weeks of age. For each animal, longitudinal strain, ejection fraction, and fractional shortening was quantified. RESULTS/ANTICIPATED RESULTS: Echocardiographic analysis of longitudinal strain in pediatric mice at 3 and 5 weeks of age is both feasible and reproducible. When compared to saline control, doxorubicin exposure at all three doses resulted in significant reductions in peak systolic longitudinal strain (–26 +/– 6% for control injected animals versus -15 +/– 4% for doxorubicin treated animals) Similar reductions in ejection fraction and fractional shortening were also noted in doxorubicin-treated mice. Somewhat surprisingly, no difference in cardiac function was noted across three different doses of doxorubicin, indicating that even a very low dose of doxorubicin induced cardiac dysfunction. DISCUSSION/SIGNIFICANCE OF IMPACT: Pediatric mice tolerate intraperitoneal injection with doxorubicin and serial echocardiography requiring sedation. Ongoing studies are focused on utilizing echocardiographic longitudinal strain to quantify cardiotoxicity during five weeks of acute anthracycline exposure followed by 13 weeks of recovery after discontinuation of therapy.
ANALYSIS OF MUSCULARIZED PULMONARY ARTERIES DUE TO PULMONARY ARTERIAL HYPERTENSION IN SICKLE CELL DISEASE
Truong U1, Solovey A1, Hebbel R1 1ACTS/SCTS, Eagan, MN, USA
OBJECTIVES/SPECIFIC AIMS: One of the leading causes of morbidity and mortality in sickle cell disease (SCD) is pulmonary arterial hypertension (PAH) instigated by increased levels of pro-inflammatory mediators including TNF-alpha. PAH triggers vascular remodeling of lungs that develop atypical muscularized intra-acinary arteries and may increase thickness of the muscular layer in larger arteries. It was proposed that TNF inhibitor entanercept (Enbrel®) would reduce the inflammation in lung arteries and reverse PAH. The aim of this study was to qualitatively examine muscularized vessels, measure the thickness of the muscular layer in pulmonary arteries, and observe the potential role of Enbrel. METHODS/STUDY POPULATION: Full, partially, and non-muscularized intra-acinary arteries were tallied. Lung sections from control, sickle, and hybrid sickle mice were immunohistochemically stained for smooth muscle actin, observed under a microscope, and images of arteries were collected. The muscular layer of the arteries was measured and a percent thickness was calculated. RESULTS/ANTICIPATED RESULTS: The data showed that the number of muscularized intra-acinary arteries was elevated in mice with SCD. There was no significant difference between the thicknesses of the muscular layer between control and sickle mice but a decrease for hybrid mice. Treatment with Enbrel showed a significant decrease in the number of muscularized intra-acinary arteries. DISCUSSION/SIGNIFICANCE OF IMPACT: The overall increase in the number of muscularized arteries in SCD tissue was indication of PAH. Although the percent thickness data did not confirm PAH, this was likely due to the fact that murine models have been demonstrated not to show ideal muscularization. Further study would be needed to show that Enbrel reduces muscularization before clinical trials. Treatment of PAH would help the many detrimental complications pertaining to the lungs and heart.
ANTIPSYCHOTIC BINDING TO DOPAMINE-3 AND DOPAMINE-2 RECEPTORS IN SCHIZOPHRENIA: A PET STUDY WITH [11C]-(+)-PHNO
Girgis R1, Slifstein M1, Abi-Dargham A1 1Columbia University and New York State Psychiatric Institute, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: Investigating the dopamine D3 receptor in patients was not possible until the recent development of [11C]-(+)-PHNO, a D3 preferring Positron Emission Tomography (PET) ligand. While significant evidence from preclinical research suggests that antipsychotic medications bind to the D3 receptor, data from some human studies suggest otherwise. Our aim is to measure the binding of the antipsychotic medication risperidone to the D3 receptor. METHODS/STUDY POPULATION: We will recruit antipsychotic free, medically healthy patients with schizophrenia. All subjects will undergo PET imaging with [11C]-(+)-PHNO, including a baseline scan, a second scan after one dose with the antipsychotic risperidone (at doses of 1–4 mg as clinically determined) and a third scan after 3 weeks of stable treatment with risperidone (at doses of 2–6 mg as clinically determined). For all scans, we will measure BPND, which is proportional to the product of receptor availability and tracer affinity, and use a regression model to estimate the D3R and D2R fractions of [11C]-(+)-PHNO BPND and the occupancy at D3 and D2 receptors by risperidone. RESULTS/ANTICIPATED RESULTS: We expect that risperidone will demonstrate D3 binding and that the lack of binding found in previous studies is related to receptor upregulation. DISCUSSION/SIGNIFICANCE OF IMPACT: The findings from this study have important implications for the development of new treatments for schizophrenia that are targeted at the D3 receptor, an endeavor already well underway.
APOLIPOPROTEIN B IN LOW DENSITY LIPOPROTEINS CONTRIBUTE TO NEUTROPHIL CONTROL OF PHAGOCYTOSED STAPHYLOCOCCUS AUREUS
Femling J1, Bryan C1, Garcia M1, Gresham H3,4, Hall P2 1University of New Mexico, Albuquerque, NM, USA; 2University of New Mexico, Department of Pharmaceutical Sciences, Albuquerque, NM, USA; 3University of New Mexico Health Sciences Center Research Service, Albuquerque, NM, USA; 4New Mexico VA Health Care System, Albuquerque, NM, USA
OBJECTIVES/SPECIFIC AIMS: Staphylococcus aureus colonizes 20–30% of the population and causes invasive deadly infections. This dual nature is due in part to the differential expression of multiple virulence factors. S. aureus virulence is controlled by density dependent quorum signaling mediated by an auto-inducing thiolactone peptide pheromone (AIP) activation of the accessory gene regulator (agr). Apolipoprotein B (apoB) is a >515 kDa amphipathic lipid binding protein and major structural protein of low density lipoprotein (LDL). ApoB inhibits AIP mediated quorum signaling and disrupts the activation of agr leading to decreased production of virulence factors. Neutrophils are crucial to the control of S. aureus, but are susceptible to lysis by ingested virulent bacteria in an agr dependent manner. The purpose of this study was to determine if apoB in LDL could contribute to neutrophil control of ingested S. aureus. METHODS/STUDY POPULATION: Neutrophils were purified from healthy human donors and exposed to S. aureus in the presence and absence of purified apoB, LDL, and oxidized LDL. After incubation both bacterial and neutrophil viability were assessed. Previously described agr deletion mutants were also compared. RESULTS/ANTICIPATED RESULTS: Purified apoB enhances neutrophil killing of ingested wild type S. aureus, but not an agr deletion mutant. ApoB and Oxidized LDL both lead to decreased S. aureus mediated neutrophil lysis. DISCUSSION/SIGNIFICANCE OF IMPACT: ApoB contributes to neutrophil mediated control of ingested S. aureus. These findings reveal a novel contribution of lipoproteins to phagocyte mediated control of S. aureus.
ARTEMETHER AND STATIN ADJUNCTIVE THERAPY ELIMINATES MORTALITY ASSOCIATED WITH EXPERIMENTAL CEREBRAL MALARIA
Wilson N1, Solomon W1, Anderson L1, Patrickson J1, Pitts S1, Bond V1, Liu M1, Stiles J1 1Morehouse School of Medicine, Atlanta, GA, USA
OBJECTIVES/SPECIFIC AIMS: Despite appropriate anti-malaria treatment, cerebral malaria-associated mortalities remain as high as 30% while 25% of survivors experience neurological complications. Excess production of Interferon gamma induced protein 10 (CXCL10) has been shown to be associated with fatal CM indicating the importance of CXCL10 in the pathogenesis of CM. However, the direct effect of increased CXCL10 production on brain cells is unknown. We assessed apoptotic effects of CXCL10 on human brain microvascular endothelial cells (HBVECs) and neuroglia cells in vitro. We tested the hypothesis that reducing excess production of CXCL10 with a synthetic drug during CM pathogenesis will reduce mortality. METHODS/STUDY POPULATION: We utilized Atorvastatin (AVA) to determine the effects of AVA and artemether (ATA) combination therapy on ECM outcome. We assessed the effect of AVA treatment on immune determinants of severity, survival, and parasitemia in ECM mice receiving a combination therapy from onset of ECM (day 6 through 9 post-infection) and comparing results with controls. RESULTS/ANTICIPATED RESULTS: The results indicate that CXCL10 induces apoptosis in HBVECs and neuroglia in a dose-dependent manner suggesting that increased level of CXCL10 in CM patients leads to neuropathogenesis and brain injury associated with CM. Treatment of ECM in mice with AVA significantly reduced systemic and brain inflammation by reducing levels of the angiostatic and apoptotic factor (CXCL10) and increasing angiogenic factor (VEGF) production. Treatment with a combination of AVA and ATA improved survival (100%) when compared with ATA monotherapy (70%), p < 0.05. DISCUSSION/SIGNIFICANCE OF IMPACT: Thus, adjunctively reducing CXCL10 levels and inflammation by AVA treatment during anti-malarial therapy may represent a novel approach to treating CM patients.
AUTOPHAGY IS A KEY FEATURE IN THE PATHOGENESIS OF SYSTEMIC SCLEROSIS (SSC)
Frech T1, De Domenico I1, Murtaugh M1, Revelo MP1, Li D1, Drakos S1 1University of Utah, Salt Lake, UT, USA
OBJECTIVES/SPECIFIC AIMS: Autophagosomes are formed during autophagy, which is activated by hypoxia and starvation. Autophagy is important for mast cell degranulation. We hypothesized that autophagy is a key feature in the pathogenesis of SSc. We examined SSc clinical features and mast cell density across the presence and severity of autophagy. METHODS/STUDY POPULATION: Skin punch biopsy was performed on 33 SSc patients and 6 healthy controls (HC) at the University of Utah. Autophagy was evaluated by immunofluorescence on paraffin sections using LC3-FITC staining on all of these patients. The intensity of staining and mast cell density was examined across clinical features in 19 of the SSc patients. Presence of autophagosome formation was assessed by EM in 17 of the SSc patients and 4 HC. RESULTS/ANTICIPATED RESULTS: In our SSc study population, 29 of subjects were female and 23 were limited cutaneous. Thirty-two of 29 SSc patients had autophagy identified by LC3-FITC staining. Intensity of staining decreased with longer duration of SSc (p = 0.09) and RP (p = 0.10). Bloating and distention differed across level of intensity staining (wilcoxin signed rank test, p = 0.05), with the greatest levels among those with moderate intensity. On EM, autophagosome formation was present in 16 of 17 SSc patients and no HC. All SSc patients had perivascular mast cells. DISCUSSION/SIGNIFICANCE OF IMPACT: Autophagy was present in 29 of 33 SSc patients and none of our healthy controls suggesting importance in pathogenesis. Autophagy staining was greater among those with shorter duration of SSc. Bloating and distention were higher in patients with moderate autophagy staining. Perivascular mast cells were present in all SSc patients. The role of autophagy in vasculopathy and mast cell activation in SSc warrants further studies.
BALANCE DEFICITS IN DIABETIC PERIPHERAL NEUROPATHY THE ROLE OF THE VESTIBULAR SYSTEM
D'Silva LJ1, Kluding PM1 1University of Kansas Medical Center, Kansas City, KS, USA
OBJECTIVES/SPECIFIC AIMS: The purpose of this study is to assess the response of people with diabetic peripheral neuropathy (DPN) to clinical tests of vestibular function. We will describe differences based on complaints of dizziness. METHODS/STUDY POPULATION: 16 subjects (mean age 58 ± 4.4) with a diagnosis of DPN completed the Functional gait Assessment (FGA), Dizziness Handicap Inventory (DHI) and Activity Specific Balance Confidence scale (ABC). RESULTS/ANTICIPATED RESULTS: Of the 16 subjects, 11 were in the mild category of the Dizziness Handicap Inventory, 4 were in the moderate category and 1 in the severe category. Analysis of the subscales of the DHI showed that dizziness affected the functional component most, followed by physical and emotional components. The mean FGA score of the 5 subjects in the moderate/severe category was 20.8 ± 2.86 which is below the fall risk cutoff of 22, while the mean score on the ABC questionnaire was 45.87 ± 21.48. Of interest is that of the 11 subjects with no/mild complaints of dizziness, the mean FGA score was 21.9 ± 4.0, indicating that they are at higher risk for falls (fall risk cutoff < 22). DISCUSSION/SIGNIFICANCE OF IMPACT: People with DPN with moderate scores on the DHI perform poorly on the FGA test. In addition, people without a history of self-reported dizziness have balance deficits as noted on the FGA score. This finding could be due to subclinical vestibular dysfunction or peripheral neuropathy. Clinicians in Physical Therapy practice frequently encounter patients with diabetes who complain of balance problems and falls especially on uneven surfaces and in the dark. These are daily situations where the vestibular system contributes significantly to prevent falls. Thorough evaluation of the vestibular system and balance deficits will guide clinicians in planning treatment interventions that include training on uneven surfaces and in altered visual situations.
BASELINE NOCICEPTION IS ALTERED IN AUTISTIC MICE
Guth K2,1, Wang L2,1, Kenyon N2,1, MIddleton L2,1, Quezado Z2,1 1Children's National Medical Center, Washignton, DC, USA; 2The Sheikh Zayed Institute for Pediatric Surgical Innovation, Washington, DC, USA
OBJECTIVES/SPECIFIC AIMS: Abnormalities in pain sensitivity are widely reported in children with autism. However, the evidence of altered pain sensitivity relies primarily on parental reporting and observations of self-injurious behavior. In order to evaluate nociception in available mouse models of autism, we examined nociception in the BTBR T+tf/J (BTBR) and the Fmr1 knock-out, a model of Fragile-X Syndrome. METHODS/STUDY POPULATION: We used the hotplate test to evaluate response to heat and the cold plate to examine response to cold. We also used a nociceptive assay that interrogates specific sensory nerve fibers. Using sine-wave electrical stimuli at 2000, 250, and 5 Hz we preferentially interrogate and measure response thresholds of Aβ, Aδ, and C sensory nerve fibers respectively. Vocalization is the end-point pain-avoiding behavior used with this nociceptive assay. RESULTS/ANTICIPATED RESULTS: We found that male and female BTBR mouse had significantly higher hot plate latency (increased tolerance to heat) and higher tolerance to cold (both, p < 0.05) compared to C57Bl/6 mice. In addition, BTBR mice had significantly higher vocalization (pain-avoiding behavior) threshold with stimulation of Aβ, Aδ, and C sensory nerve fibers compared to C57Bl/6 (all p < 0.05). Fmr1 KO male mice, in contrast, had significantly higher hotplate paw withdrawal latency (p < 0.05), but similar cold and sensory fibers response to electrical stimulation compared to the FVB/NJ mice. DISCUSSION/SIGNIFICANCE OF IMPACT: Our findings suggest in mouse models of autism baseline nociception is altered. Extrapolating our findings clinically, these data support the notion that nociception might be altered in autistic individuals and that the nature of this alteration might vary according to the etiology of autism
BEHAVIORAL CHARACTERIZATION AND PHARMACOLOGICAL TREATMENT OF AN INBRED MOUSE MODEL OF RESTRICTED, REPETITIVE BEHAVIOR
Muehlmann A1, Lewis MH1 1University of Florida, Gainesville, FL, USA
OBJECTIVES/SPECIFIC AIMS: Clinical and animal studies have provided only very limited findings with respect to the genes or neuropathology that control restricted, repetitive behavior (RRB) in neurodevelopmental disorders. In order to investigate the genetics and neuropathological basis of RRB, we have further characterized the expression of the restricted, repetitive behavioral phenotype reported in the C58 inbred mouse strain. METHODS/STUDY POPULATION: We assessed repetitive motor behavior across the 12 hour dark cycle using automated apparatus and video-recording. We used a holeboard exploration task to assess restricted behavior and the marble-burying task to assess perseverative motor responding. RESULTS/ANTICIPATED RESULTS: C58 mice displayed high levels of spontaneous repetitive motor behavior (e.g. vertical jumping and backward somersaulting). No significant strain differences were noted in the holeboard exploration task. C58 mice engaged in less marble-burying than C57BL/6 mice and marble-burying was significantly inversely correlated with stereotyped motor behavior. These findings confirmed RRB as a robust, quantifiable, and reliable behavioral phenotype in this strain. Furthermore, we identified that the RRB in C58 mice is associated with hypofunctioning of the indirect basal ganglia pathway and can be reduced by a drug cocktail that targets indirect pathway neurons in the striatum. The triple combination of dopamine D2 antagonist, adenosine A2a agonist, and glutamate mGluR5 positive allosteric modulator significantly reduced RRB better than any single or double drug combination. DISCUSSION/SIGNIFICANCE OF IMPACT: These studies indicate that the C58 mouse model has important translational value in ascertaining the genetics, neuropathology, and drug treatment of RRB in autism and other neurodevelopmental disorders.
BIOMARKER IDENTIFICATION OF NUCLEAR MAGNETIC RESONANCE (NMR)-BASED EXHALED BREATH CONDENSATE USING MULTIVARIATE STATISTICAL ANALYSIS
Fulcher YG1, Fotso M1, Chang C2, Reinero C2, Van Doren SR1 1University of Missouri-Columbia, Columbia, MO, USA; 2Veterinary Medicine & Surgery Department, Univerisity of Missouri-Columbia, Columbia, MO, USA
OBJECTIVES/SPECIFIC AIMS: With the prevalence of asthma and imprecise methods of clinical diagnosis, there is a clear need for identifying accurate and significant biomarkers for asthma diagnosis. Exploration for non-invasive disease-specific biomarkers to assess the health of the lung for early detection of asthma can be realized through collecting Exhaled Breath Condensate (EBC) and Nuclear Magnetic Resonance (NMR)-derived metabonomic data analysis. METHODS/STUDY POPULATION: The population of this study included 33 healthy cats and 34 cats induced to have mild asthma. 7 of the asthmatic cats were treated with Prednisolone. The NMR spectra of the exhaled breath condensate from the cats were collected. Multivariate statistical analyses such as principle component analysis (PCA) and partial-least square discriminant analysis (PLS-DA) were examined to compare the EBC sample components of healthy, asthmatic and steroid-treated asthmatic cats. RESULTS/ANTICIPATED RESULTS: PCA and PLS-DA successfully distinguished healthy cats from asthmatic cats. Several NMR peaks that statistically distinguish EBC samples of healthy vs. asthmatic cats were identified. Two NMR Peaks matched to lactic acids, identifying its loss as a biomarker of mild asthma in cats. DISCUSSION/SIGNIFICANCE OF IMPACT: NMR-based EBC fingerprinting discriminated healthy cats from asthmatic cats and identified the metabolites that may be responsible for the difference. Identification of biomarkers of asthma in cats could place them on the “watch list” for monitoring by the most convenient method of analytical chemistry and for evaluation as potential biomarkers of human asthma.
BIOMARKERS FOR NEURONAL AND IMMUNE MECHANISMS IN COMBAT PTSD: PILOT RESULTS
Lapiz-Bluhm M1, Williamson D1, Peterson A1 1University of Texas Health Science Center at San Antonio, San Antonio, TX, USA
OBJECTIVES/SPECIFIC AIMS: Post-traumatic stress disorder (PTSD) is an anxiety disorder characterized by symptoms of re-experiencing, avoidance and increased arousal following exposure to a traumatic event. Little progress has been made towards identifying reliable diagnostic biomarkers in PTSD. This study determined the levels of proteins associated with neuronal plasticity and immune function in active Military members with or without PTSD symptoms. The proteins studied were brain-derived neurotrophic factor (BDNF), platelet-derived growth factor (PDGF) AA, PDGF AB/BB, cathepsin D, myeloperoxidase (MPO), regulated on activation normal T expressed and secreted (RANTES) interleukin, neural cell adhesion molecule (NCAM), soluble intercellular adhesion molecule 1 (sICAM-1), and plasminogen activator inhibitor 1 soluble vascular adhesion molecule (sVCAM-1) . METHODS/STUDY POPULATION: Plasma samples from 38 male Caucasian active military members with or without PTSD symptoms (based on their PTSD Checklist-Civilian (PCL-C) scores) were assayed for BDNF, PDGF AA, PDGF AB/BB, cathepsin D, MPO, RANTES, NCAM, sICAM-1 and sVCAM-1 using the MILLIPLEX® MAP Bead Panel and Luminex Flex Map3D. Protein levels were expressed as net mfi and analyzed for statistical significance (p < 0.05). RESULTS/ANTICIPATED RESULTS: Individuals with high PCL-C scores showed significantly higher (p < 0.05) RANTES levels compared to those with low PCL-C scores. No significant difference detected between groups for BDNF, PDGF AA, PDGF AB/BB, cathepsin D, MPO, NCAM, sICAM-1) and sVCAM-1 levels. DISCUSSION/SIGNIFICANCE OF IMPACT: This pilot study is first to document the differential level of RANTES, a member of the cytokine-8 family, in PTSD symptomatology. The involvement of this immune pathway may potentially be targeted in interventions for PTSD. Larger studies, possibly with a cohort of normal civilian controls, are planned.
BLOOD OXYGENATION LEVEL-DEPENDENT MAGNETIC RESONANCE IMAGING IN SKELETAL MUSCLE AT 7 TESLA
Towse TF1,2, Sabin SA1, Bush EC1, Childs BT1, Damon BM1 1Vanderbilt University, Nashville, TN, USA; 2Physical Medicine and Rehabilitation, Nashville, TN, USA
OBJECTIVES/SPECIFIC AIMS: Blood oxygenation level-dependent (BOLD) magnetic resonance imaging (MRI) may provide a non-invasive technique for measuring peripheral vascular (PV) function. BOLD based imaging, coupled with brief muscle contractions, results in an increase in the contrast of an MR image and is called a BOLD response. The biophysical basis of the BOLD response, at MR field strengths of 1.5 and 3 Tesla (T), originates from changes in blood volume and blood oxygenation in the tissue. At higher field strengths the BOLD response may also be sensitive to structural aspects of the vascular anatomy. BOLD based imaging has already been used to assess PV function in obese non-diabetic, and Type 1, and Type 2 diabetic patients but only at 1.5 and 3T. The purpose of this project is to determine the feasibility muscle BOLD imaging at 7T. METHODS/STUDY POPULATION: Two subjects (1 male) aged 30.5 ± 12.0 yrs., BMI = 28.9 ± 0.4 (mean ± SD), participated in the study. Subjects were positioned supine in a 7T Philips scanner with the right calf muscle positioned in the center of an extremity coil. During the MRI scan BOLD sensitive images were acquired continuously for 7 mins. The subject performed a single, brief maximal contraction of the plantar flexor muscles against a resistance band 30 sec into the scan. The protocol was repeated 3–4 times for each subject. RESULTS/ANTICIPATED RESULTS: There was a detectable BOLD response in the calf muscles of each subject following the contractions. The magnitude and timing of the response is consistent with the BOLD response seen at lower field strengths. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results suggest muscle BOLD imaging is feasible at 7T. Future studies will determine the biophysical basis of the BOLD response at 7T and apply this technique in clinical populations.
CALORIC RESTRICTION MODULATES CA1 PYRAMIDAL NEURON GENE EXPRESSION AND REDUCES AMYLOID BURDEN IN AN ALZHEIMER'S DISEASE MOUSE MODEL
Schafer M1,2, Ginsberg S1,2 1New York University, New York, NY, USA; 2Nathan Kline Institute, Orangeburg, NY, USA
OBJECTIVES/SPECIFIC AIMS: Alzheimer's disease (AD) is a neurodegenerative disorder characterized by age-dependent amyloid-beta (Aβ) peptide aggregation, neurofibrillary tangles deposition, cell-type specific vulnerability, and cognitive deficits. Caloric restriction (CR) is a dietary intervention that may reduce AD pathology. Previous studies have demonstrated CR-induced pathology reduction through regional, low-resolution analysis. CR benefits in vulnerable hippocampal CA1 pyramidal neurons have not been characterized. METHODS/STUDY POPULATION: Using laser capture microdissection and microarray analysis, we are investigating gene expression profiles of CA1 neurons isolated at time points corresponding to pathology onset and later progression (5 and 15 mos.) from the Tg2576 model of cerebral amyloidosis and nontransgenic (ntg) littermates maintained on 30% CR versus ad libitum (AL) feeding. RESULTS/ANTICIPATED RESULTS: Preliminary results of 5 mo. old mice indicate that CR decreases levels of several classes of relevant transcripts, including genes implicated in transcription, autophagy, and metabolism, while selectively increasing synaptic plasticity genes. 5 and 15 mo. old transcription profile comparisons indicate that CR attenuates the number of genes that are downregulated throughout aging, demonstrating that CR may blunt aging mechanisms. Consistent with previously published results, quantitative assessments indicate that CR-Tg2576 mice contain significantly less Aβ40 and Aβ42 protein than AL-Tg2576 mice, demonstrating functional pathology reduction. DISCUSSION/SIGNIFICANCE OF IMPACT: We expect additional analyses to confirm that CR-induced expression level changes are associated with upregulation of neuroprotective pathways, contributing to reduced pathology that may have translational potential for AD remediation in at risk human populations.
CD90+ COLONIC STROMAL CELLS: ROLE IN THE IMMUNOPATHOGENESIS OF ULCERATIVE COLITIS
Pinchuk I1, Beswick EJ2, Morris K2, Saada JI1, Warwick J1, Powell DW1, Reyes V1 1UTMB, Galveston, TX, USA; 2University of New Mexico, Albuquerque, NM, USA
OBJECTIVES/SPECIFIC AIMS: Underlying mechanisms of dysregulation of CD4+ T cell responses resulting in pathological Th2/Th17 responses during ulcerative colitis (UC) are unclear. Colonic mucosal CD90+ stromal cells (a.k.a myofibroblasts/fibroblasts, CMFs) isolated from normal mucosa (N-CMFs) suppress IFN-γ, a Th1 cytokine, via PD-L1-dependent mechanism. Increase in expression of PD-L1 and a Th17 promoting cytokine IL-6 was noted in UC-CMFs when compared to N-CMFs. We hypothesized that upregulation of PD-L1 and IL-6 expression by UC-CMFs contributes to the Th2/Th17 type inflammatory responses involved in UC immunopathogenesis. METHODS/STUDY POPULATION: Flow cytometry and Luminex arrays were used to analyze phenotype of UC-CMF and regulation CD4+ T cell responses by CMFs. RESULTS/ANTICIPATED RESULTS: Flow cytometry analysis of colonic mucosal single cell (CM) preparations demonstrated that PD-L1 is upregulated in colonic mucosa. CD90+ CMFs were the major cell phenotype expressing PD-L1 in CM preparations. UC-CMF maintained elevated level of PD-L1 expression in culture and produced increased levels of IL-6 when compared to N-CMFs and caused stronger suppression of IFN-γ+Tbet+ Th1 type cells. This suppression was reversed in presence of anti-PD-L1 blocking antibodies. In contrast to N-CMFs, priming of CD2/CD3/CD28-activated naïve Th cells with UC-CMFs enhanced generation of the Th cells expressing the Th17 transcriptional factor RORγτ in IL-6 dependent manner. DISCUSSION/SIGNIFICANCE OF IMPACT: Our data suggest that during chronic inflammation UC-CMFs have stable alteration of their phenotype. Further, via upregulation of PD-L1, which inhibit Th1 and, thus, shift balance to Th2 type responses and increase in IL-6, a Th17 promoting cytokine, UC-CMFs may be involved in the Th2/Th17 chronic inflammatory responses during immunopathogenesis of UC.
CHANGES IN THE IGF AXIS AMONG ELDERLY DIABETICS AND NON-DIABETICS
Aneke C1, Parrinello C2, Rajpathak S3,4, Rohan T1, Strotmeyer E4, Kritchevsky S5, Psaty B6, Buzkova P5, Kizer J7, Newman A4, Strickler H1, Kaplan R1 1Albert Einstein College of Medicine, Bronx, NY, USA; 2Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA; 3Perelman School of Medicine at University of Pennsylvania, Philadelphia, PA, USA; 4University of Pittsburgh, Pittsburgh, PA, USA; 5Wake Forest University School of Medicine, Winston-Salem, NC, USA; 6University of Washington, Seattle, WA, USA; 7Weill Medical College of Cornell University, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: IGF-I is structurally similar to insulin and little is known about long term changes in IGF-I and IGFBPs in normoglycemics who later develop IGT, IFG, both IGT and IFG or diabetes. The aim is to assess whether changes in IGF-I, IGFBP-1 and IGFBP-3 protein levels are more prominent in those with worsening versus stable glycemic status over a 9-year follow up period. METHODS/STUDY POPULATION: We designed a retrospective analysis of CHS All Stars, a cohort study in older adults aged over 65 years of age. ADA criteria for IGT, IFG, and diabetes were used to classify participants. RESULTS/ANTICIPATED RESULTS: Mean IGF-I and IGFBP-3 levels decreased while mean IGFBP-1 levels increased during follow up. The greatest change in all protein levels occurred in those who were diabetic at baseline and the smallest change in those who were normoglycemic at baseline and follow up. DISCUSSION/SIGNIFICANCE OF IMPACT: This suggests that changes in the IGF-axis vary for those with different glycemic profiles, which may lead to different health outcomes in diabetics compared to non-diabetics.
CHARACTERIZATION OF HANGOVER FOLLOWING INTRAVENOUS (IV) ALCOHOL IN SOCIAL DRINKERS: EFFECTS OF FAMILY HISTORY, DRINKING HISTORY AND SEX
Schmidt VY1, Vatsalya V1, Ramchandani VA1 1NIH/NIAAA, Bethesda, MD, USA
OBJECTIVES/SPECIFIC AIMS: Hangover is a common alcohol-related consequence associated with significant health and socioeconomic impact. Research on hangover is limited and it is unclear if studies using IV alcohol result in hangover symptoms. This study aimed to characterize hangover following IV alcohol, and effects of family history, drinking history and gender, in social drinkers. METHODS/STUDY POPULATION: 23 healthy social drinkers (11 F) participated in a 2-session study, and received, in separate sessions, IV alcohol to achieve 2 exposure profiles: an alcohol clamp and a biphasic profile that mimicked a standard oral alcohol exposure. Acute Hangover Scale (AHS) was used to assess hangover symptoms and post-infusion drinking from the end of the session until the next morning. Family tree questionnaire was used to assess family history of alcoholism (FHA), and lifetime and recent drinking history were also assessed. RESULTS/ANTICIPATED RESULTS: Results indicated that 77% of subjects reported hangover symptoms, with mild to moderate severity for the items “tired”, “thirsty” and “headache”. AHS scores showed a significant Sex effect and FHA X Sex interaction, with FHA females reporting the highest symptoms, after controlling for drinking history. AHS scores did not differ between clamp and biphasic profiles. The proportion of subjects reporting post-infusion drinking showed significant effects of recent drinking, (average drinks per day and heavy drinking days) during the clamp session. DISCUSSION/SIGNIFICANCE OF IMPACT: This study demonstrates, for the first time, that IV alcohol is associated with mild to moderate hangover symptoms in social drinkers. Females, particularly those with FHA, showed greater hangover symptoms than males.
CHITOTRIOSIDASE AND IMMUNOGLOBULIN E PREDICT AIRWAY OBSTRUCTION IN WORLD TRADE CENTER EXPOSED NEW YORK CITY FIREFIGHTERS
Cho S1, Echevarria G1, Kwon S1, Schenck E1, Tsukiji J1, Prezant DJ2, Rom WN1, Nolan A1, Weiden MD1,2 1NYU, New York, NY, USA; 2Fire Department of New York, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: World Trade Center (WTC) exposure caused airflow obstruction in FDNY firefighters. Serum biomarkers are risk factors for future forced expiratory volume at 1 second (FEV1) decline. Biomarkers of abnormal FEV1/Forced Vital Capacity (FVC) in this cohort are unknown. We investigated if serum chitotriosidase and IgE early after WTC exposure can predict subsequent abnormal FEV1/FVC. METHODS/STUDY POPULATION: In a nested case control study we compared chitotriosidase and IgE in serum samples drawn within 6 months of 9/11 with low FEV1/FVC defining obstruction at subspecialty pulmonary evaluation (SPE) performed years later. We studied 251 WTC-exposed never smoking firefighters without pre-9/11 lung disease. Cases (N = 125) had FEV1/FVC less than lower limit of normal (LLN) at SPE whereas controls had normal FEV1/FVC (N = 126). Immunoassay measured serum biomarkers and logistic regression modeled that association between biomarkers and disease. RESULTS/ANTICIPATED RESULTS: Cases had lower serum chitotriosidase and higher IgE than controls. Each 10ng/ml increase in chitotriosidase decreased the odds of abnormal FEV1/FVC at SPE by 12.7% (95%CI:0.71–0.95) while each 100 IU/mL increase in IgE increases the odds of airflow obstruction by 24.7% (95%CI:1.09–1.72). The multivariable model was adjusted for age, BMI, exposure group and baseline FEV1/FVC. The area under the receiver operating characteristic curve was 0.86. DISCUSSION/SIGNIFICANCE OF IMPACT: Increased serum chitotriosidase reduces the risk of developing abnormal FEV1/FVC after WTC particulate matter exposure while elevated IgE was a risk factor for airflow obstruction. Both innate and adaptive immune mediators independently predict future FEV1/FVC ratio in our FDNY cohort after WTC PM exposure.
CHRONIC MTOR ACTIVATION PROMOTES CELL SURVIVAL IN MERKEL CELL CARCINOMA
Gao L1, Lin Z1, McDermott A2, Shao L3, Kannan A1, Stack BC4, Moreno M4, Davis D1, Cornelius LA5 1Department of Dermatology,University of Arkansas for Medical Sciences (UAMS), Little Rock, AR, USA; 2College of Medicine, UAMS, Little Rock, AR, USA; 3Department of Otolaryngology-Head and Neck Surgery, UAMS, Little Rock, AR, USA; 4Division of Dermatology, Washington University School of Medicine in St. Louis, St. Louis, MO, USA; 5Division of Radiation Health, Department of Pharmaceutical Sciences, UAMS, Little Rock, AR, USA
OBJECTIVES/SPECIFIC AIMS: Merkel cell carcinoma (MCC) is an aggressive skin cancer with rising incidence. So far no evidence-based chemotherapy exists. Accumulating evidence indicates aberrant mTOR pathway and autophagy in many human cancers. However, no detailed investigation of mTOR pathway in MCC has been performed. METHODS/STUDY POPULATION: To evaluate mTOR pathway and autophagy, we employed tissue microarray consisting of 65 MCC samples. Our findings were then confirmed using 9 fresh MCC tumors. We further assessed the biologic effects of mTOR inhibition in a primary human Merkel cell carcinoma cell line. RESULTS/ANTICIPATED RESULTS: We have demonstrated that mTOR activation and suppressed autophagy is common in MCCs regardless of the presence Merkel cell carcinoma virus (MCV). Our data also suggests alternative mechanisms of mTOR activation. We have shown the specific inhibition of mTOR pathway induces autophagy and cell death in a human primary Merkel cell carcinoma cell line. Furthermore, increased cell death induced by mTOR inhibitors is independent of caspase activation but can be attenuated by autophagy inhibitor. DISCUSSION/SIGNIFICANCE OF IMPACT: This is the first study to evaluate mTOR pathway and autophagy thoroughly in MCC. Our data suggests a potential role of autophagic cell death upon inhibition of mTOR pathway. Our observations have thus uncovered a previously underappreciated role of mTOR signaling and cell survival, and merit further studies for potential therapeutic targets in MCC management.
COMBINATION OF CIGARETTE SMOKE EXPOSURE AND OVEREXPRESSION OF THE a-EPITHELIAL NA+ CHANNEL MIMICS GENE EXPRESSION CHANGES FOUND IN HUMAN CHRONIC OBSTRUCTIVE PULMONARY DISEASE
Duncan EA1, Prabhu S1, Livraghi-Butrico A1, Dang H1, Tilley SL1, Boucher RC1, O'Neal WK1, Doerschuk CM1 1University of North Carolina at Chapel Hill, Chapel Hill, NC, USA
OBJECTIVES/SPECIFIC AIMS: Mouse models that mimic human chronic obstructive pulmonary disease (COPD) are lacking. Transgenic mice that overexpress the β-epithelial Na+ channel (βENaC) develop features of COPD, but lack exposure to cigarette smoke. We sought to determine whether βENaC overexpression, cigarette smoke exposure, or the combination of both cause changes in gene expression patterns that mimic those in human COPD. METHODS/STUDY POPULATION: Wild type (WT) and βENaC-overexpressing mice were exposed to 6 months of cigarette smoke or sham air. Alveolar destruction and airway mucus accumulation were analyzed. Lung gene expression was analyzed using microarrays and compared to that in public databases from humans with COPD. RESULTS/ANTICIPATED RESULTS: In WT mice, cigarette smoke exposure caused mild emphysema and no airway mucus accumulation. βENaC-transgenic mice had more severe emphysema and airway mucus accumulation than smoke-exposed WT mice, neither of which were effected by cigarette smoke. Gene set enrichment analysis revealed that βENaC overexpression led to up-regulation of some immune regulatory pathways. Cigarette smoke exposure in these mice led to up-regulation of oxidative stress response and detoxification pathways and down-regulation of a partially overlapping set of immune regulatory pathways. Modulation of both gene expression patterns were found in published gene expression data in human COPD. DISCUSSION/SIGNIFICANCE OF IMPACT: In mice, cigarette smoke exposure and βENaC overexpression lead to gene expression changes consistent with human COPD, suggesting this model may be helpful in studying the pathogenesis of COPD and evaluating therapeutic interventions.
COMBINED RADIOTHERAPY AND IMMUNOTHERAPY USING CPG OLIGODEOXYNUCLEOTIDES AND INDOLAMINE 2,3 DIOXYGENASE (IDO) BLOCKADE
Monjazeb AM1, Kent M2, Sckisel G2, Sparger E2, Murphy WJ2 1UC Davis, Sacramento, CA, CA, USA; 2UC Davis, Davis, CA, USA
OBJECTIVES/SPECIFIC AIMS: Localized radiotherapy (RT) can induce a systemic anti-tumor immune response. Two clinical trials demonstrate the effectiveness of combining RT and CpG immunotherapy. We test the efficacy and safety combining RT and CpG with blockade of the immunosuppressive enzyme, IDO, using 1-Methyl D-Tryptophan (1-MT). METHODS/STUDY POPULATION: Tumor bearing mice were treated with RT + IT. IT consisted of intratumoral CpG and 1-MT administered in the drinking water. Tumor growth, survival, toxicity and the immune profile of various tissues were assessed. A similar clinical trial was initiated at the UC Davis Veterinary Cancer Center testing this approach for spontaneous metastatic cancers in companion canines. RESULTS/ANTICIPATED RESULTS: In mice therapy improved overall survival, decreased lung metastases, decreased mean tumor growth and increased percentage of activated dendritic cells in tumor draining lymph nodes. No treatment related toxicities were observed. In the canine clinical trial 3 patients have been enrolled, 1 of which has completed therapy to date. No treatment related toxicities have been observed. In the canine which has completed therapy (Alaskan Husky with Metastatic Melanoma) regression of the primary tumor and stability of lung metastases has been observed at 3 months post-treatment. The treatment has drastically reduced the level of regulatory T-cells in the tumor, draining lymph nodes, and peripheral blood. DISCUSSION/SIGNIFICANCE OF IMPACT: Combining RT and IT is more effective than either therapy alone. Preliminary results in spontaneous canine cancers also show promise. Further study is needed to determine the immunologic mechanism of this therapy. This therapy has limited toxicity in both mice and canines and is being considered for translation to human studies.
CONJUGATING POLY(ETHYLENE) GLYCOL (PEGYLATION) TO A HIGH-AFFINITY ANTI-METHAMPEHTAMINE ANTIBODY FRAGMENT
Reichard EE1, Gonzalez GA1, Peterson EC1 1University of Arkansas for Medical Sciences, Little Rock, AR, USA
OBJECTIVES/SPECIFIC AIMS: Methamphetamine (METH) is considered one of the top drug problems in the United States today, yet there are no FDA approved pharmacological treatments available for METH abuse. To this end, we have designed and produced an anti-METH single chain antibody fragment (scFv7F9Cys) as a potential anti-METH pharmacological treatment. However, scFv7F9Cys has a short half-life in its native state, limiting its clinical use. Thus, the aim of this study was to examine the in vitro effects of conjugating scFv7F9Cys to poly(ethylene) glycol (PEG) as a potential method of increasing half-life. METHODS/STUDY POPULATION: Three different PEG moieties were tested: linear 5 kDa and 20 kDa PEGs and a branched 40 kDa PEG. ScFv7F9Cys was conjugated to each PEG forming three different scFv7F9Cys-PEG conjugates (scFv7F9Cys-PEG5K, -PEG20K, or -PEG40K). ScFv7F9Cys-PEG conjugates were then purified from unreacted scFv7F9Cys and PEG using affinity chromatography (IMAC). PEGylation and purification were analyzed by SDS-PAGE and size exclusion chromatography. METH binding affinity and capacity were tested for each scFv7F9Cys-PEG conjugate and compared to native scFv7F9Cys using saturation binding assays. RESULTS/ANTICIPATED RESULTS: Both PEGylation of scFv7F9Cys and purification of scFv7F9Cys-PEG were successful and binding analysis suggests PEGylation did not significantly alter METH affinity or binding capacity. DISCUSSION/SIGNIFICANCE OF IMPACT: We believe successful PEGylation of scFv7F9Cys, while maintaining METH binding capabilities, will produce a clinically relevant pharmacotherapy for the treatment of METH addiction, useful in both acute/overdose scenarios and long-lasting relapse prevention. Funding: NIDA R01 DA026423, NIH CSTA 1UL1RR029884, NIDA T32 DA022981.
CREATING A SURGICAL ALVEOLAR CLEFT MODEL IN JUVENILE SWINE TO TEST NOVEL AUTOLOGOUS TREATMENT STRATEGIES
Halevi AE1, Emodi O1, Skancke M1, Hill J1, Caballero M1, Pharaon M1, van Aalst JA1 1University of North Carolina, Chapel Hill, NC, USA
OBJECTIVES/SPECIFIC AIMS: Craniofacial bone defects occur in 1 in 250 live births. To generate new treatment algorithms for these defects, large animal models are needed. Swine, like humans, are omnivores, with similar size and function of the facial skeleton, making them ideal for this work. This study develops a juvenile swine alveolar cleft model to test autologous tissue engineered bone. METHODS/STUDY POPULATION: To refine technique 4 swine cadavers were utilized. In addition, 4 one-month-old swine underwent surgical alveolar cleft creation to determine maxillary critical-sized defects and healing after rib or iliac crest cancellous bone grafts. Radiographs, computed tomography (CT) scans, and micro-CTs were obtained at initial surgery and 1- month sacrifice. Harvested bone was evaluated histologically and with 4-point mechanical testing. RESULTS/ANTICIPATED RESULTS: In cadaver swine, 1 cm defects were surgically created in 6 hemi maxillae and treated with rib (n = 2) and iliac crest (n = 4) grafts. Bilateral 1 cm and 2 cm defects (n = 2) did not result in an unstable central maxilla. In 1-month old swine, a 1 cm surgically created alveolar cleft healed without treatment in 1 month; a 2 cm defect did not heal. Rib graft did not incorporate into surrounding bone. Cancellous bone healed in the 2 cm defect with normal histological bone. DISCUSSION/SIGNIFICANCE OF IMPACT: This pilot study establishes a juvenile swine unilateral alveolar cleft model with a 2-cm critical-sized defect. Cadaver work suggests that a bilateral defect can be created in a living animal without maxillary instability. This work establishes the foundation for a surgically created bilateral alveolar cleft model in juvenile swine to study novel autologous treatments in which to test mesenchymal stem cell treatment strategies.
DEVELOPING A MOLECULAR UNDERSTANDING OF NON-SMALL CELL LUNG CANCER (NSCLC) RESISTANCE TO STANDARD TAXANE-PLATIN CHEMOTHERAPY
Dalvi MP1, Behrens C2, Suraokar M2, Girard L1, Xie Y1, Wistuba I2, Minna JD1 1UT Southwestern Medical Center, Dallas, TX, USA; 2MD Anderson Cancer Center, Houston, TX, USA
OBJECTIVES/SPECIFIC AIMS: Resistance to standard taxane-platin doublet therapy in NSCLC is not fully understood and hence the objective of this work is to identify chemo-resistance mechanisms using pre-clinical and clinical models. METHODS/STUDY POPULATION: We used 3 complementary approaches to explore NSCLC resistance to taxane+platin chemotherapy. Cell lines were exposed in vitro to chemotherapy cycles to mirror clinical regimens. Xenografts of cell lines established in NOD/SCID mice were treated with taxane+platin. Clinically annotated patient tumors were obtained after neoadjuvant chemotherapy (N = 66) or no chemotherapy (N = 209). Genome-wide mRNA expression profiles were compared in these three models representing basic to clinical translation. RESULTS/ANTICIPATED RESULTS: NSCLC cell lines NCI-H1299 and NCI-H1355 treated in vitro for 16–18 cycles with paclitaxel+carboplatin developed >50 fold resistance to this doublet. Cells showed cross-resistance to docetaxel+cisplatin, doxorubicin and vinorelbine. Drug selected cells also showed in vivo resistance. Treated xenograft tumors showed mRNA expression differences compared to controls. Similarly, neoadjuvant treated patient tumors had differential expression profiles. Genome-wide profiles from all three settings revealed among others, multi-drug transporters, epigenetic modulators, cancer stem cell genes and cancer-testis antigens. DISCUSSION/SIGNIFICANCE OF IMPACT: We are now systematically testing this common subset of genes to determine which are directly related to drug resistance. This should provide both clinically useful biomarkers to type tumors in patients and also potentially new therapeutic targets to reverse/overcome taxane-platin drug resistance in NSCLC.
DISCOVERY OF NEW DRUG EFFLUX TRANSPORTERS ON CORNEA - MOLECULAR, FUNCTIONAL SCREENING AND DRUG-HPLC METHOD VALIDATION
Karla PK1, Oyugi D1, Ako-Adouno A1, Herndon BL2, Jones L1 1Howard University, Washington, DC, USA; 2University of Missouri, Kansas City, MO, USA
OBJECTIVES/SPECIFIC AIMS: The objective of this study is to screen the human corneal epithelial cells for molecular and functional presence of new drug efflux transporters and develop / validate specific, sensitive yet cost effective HPLC method for the identification and quantification of Timolol drug concentrations in ophthalmic nanoparticle formulations. METHODS/STUDY POPULATION: Serum Institute Rabbit Corneal Epithelial Cells (SIRC) and Transfected Human Corneal Epithelial Cells (SV40-HCEC) were employed for screening analysis. RT-PCR followed by sequencing was employed for gene expression. Flow cytometry was employed for drug-transporter screening. A Shimadzu HPLC-UV System with C18 column and a methanol:water with 0.1% formic acid (60:40 v/v) mobile phase was employed for method development. RESULTS/ANTICIPATED RESULTS: SIRC showed specific PCR bands at 101 bp, 104 bp, 106 bp, 103 bp, 114 bp, and 135 bp, corresponding to MRPs 3, 4, 5, P-gp, SIMRP-7 and BCRP, respectively. These band profiles, together with MRPs 1 (107 bp), 2(114 bp), 6 (105 bp), and ABCCs 10 (114 bp), 11(112 bp) and 12 (112 bp) were observed in SV40 HCEC. Rhodamine intensity in cells treated with MK571, P-gp 4008, Timolol, Ganciclovir (GCV), and Latanoprost (LNP) at 50 uM and 100 uM was higher than control. Timolol retention time for HPLC was 7.6 min. The LOD and LOQ were 0.21 μg/ml and 0.68 μg/ml respectively. The mean%RSD was found to be 0.17%. DISCUSSION/SIGNIFICANCE OF IMPACT: The results confirm the expression of multiple new transporters at gene level on human corneal cells. Flow-cytometry screening studies revealed that major drugs, Timolol, GCV, LNP are substrates for drug efflux. A reliable, sensitive and cost effective RP-HPLC method to determine drug (Timolol) release profiles from nano formulations was successfully developed and validated.
DOWN-REGULATION OF THE MATRICELLULAR PROTEIN, PERIOSTIN, IS A NOVEL BIOMARKER OF ACUTE LUNG INJURY
Ahlfeld SK1, Gao Y1, Conway S1 1Program in Developmental Biology, Indiana University School of Medicine, Indianapolis, IN, USA
OBJECTIVES/SPECIFIC AIMS: Extreme preterm birth exposes the saccular lung to multiple teratogens which retard alveolar lung development resulting in Bronchopulmonary Dysplasia (BPD), a chronic lung disease of preterm infants. Preventative therapies have been hindered by a lack of reliable markers of early disease pathogenesis. Up-regulation of the matricellular protein, Periostin, is a marker of late lung injury, myofibroblast accumulation, and pulmonary fibrosis but it is unknown if Periostin directs normal myofibroblast accumulation during early alveolar septation or if it identifies early lung injury. METHODS/STUDY POPULATION: Using a murine hyperoxia (85%-O2) model of inhibited alveolar septation, we correlated Periostin expression with the appearance of myofibroblasts in the saccular lung, determined if it was required for early alveolar development, and examined its misexpression in early lung injury. RESULTS/ANTICIPATED RESULTS: Periostin was widely expressed in distal lung septa through P2–4 (early alveolar septation) and peak expression coincided with accumulation of α-smooth muscle actin (αSMA)+-myofibroblasts. Acutely, hyperoxia blunted normal up-regulation of Periostin and αSMA prior to the first evidence of distal lung simplification at P4. Conversely, chronic hyperoxia resulted in abnormal accumulation of Periostin and αSMA in blunted septa at P14. Despite its tight association with hyperoxic lung injury, Periostin knockout mice exhibited normal myofibroblast differentiation and alveolar development, and they were equally sensitive to hyperoxia as wildtypes. DISCUSSION/SIGNIFICANCE OF IMPACT: We conclude that Periostin is a novel biomarker of acutely-inhibited alveolar septation during a clinically-relevant window of lung development.
DUAL TRANSCRIPTOME ANALYSIS OF EPSTEIN-BARR VIRUS (EBV) ASSOCIATED GASTRIC CARCINOMA RNA-SEQ DATA REVEALS EBV REGULATION OF GASTRIC CARCINOMA TUMOR ENVIRONMENT; IMPLICATIONS FOR POSSIBLE IMMUNE ADJUVANT THERAPY
Strong MJ1, Xu G2, Coco J2, Baribault C3, Vinay DS4, Lacey MR3, Lin Z1, Baddoo M1, Concha M1, Burow ME4, Taylor CM5, Flemington EK1 1Tulane University, Department of Pathology, New Orleans, LA, USA; 2University of New Orleans, Department of Computer Science, New Orleans, LA, USA; 3Tulane University, Department of Mathematics, New Orleans, LA, USA; 4Tulane University, Department of Medicine, New Orleans, LA, USA; 5Louisiana State University School of Medicine, Department of Microbiology, Immunology & Parasitology, New Orleans, LA, USA
OBJECTIVES/SPECIFIC AIMS: Epstein-Barr virus (EBV) is associated with roughly 10% of gastric carcinomas worldwide (EBVaGC). In this study, we investigated EBVaGC using RNA-seq data to link EBV genes (and expression intensities) to cell and microenvironmental changes. METHODS/STUDY POPULATION: Using a cohort of 71 gastric carcinoma RNA-seq data sets from the Cancer Genome Atlas, we performed a quantitative and global assessment of EBV gene expression in gastric carcinomas and assessed EBV associated cellular pathway alterations. RESULTS/ANTICIPATED RESULTS: EBV transcripts were detected in 17% of samples. In 4 samples with the highest EBV coverage (hiEBVaGC), transcripts from the BamHI A region comprised the majority of EBV reads. The 4 hiEBVaGCs displayed elevated immune cell infiltration with high interferon-gamma expression compared to samples with low or no EBV gene expression. DISCUSSION/SIGNIFICANCE OF IMPACT: Along with increased immune cells, hiEBVaGC samples showed elevated expression of the potent immune cell inhibitor, IDO1. This finding may partly explain the persistence of these virus associated tumors in the face of local immune cell concentration. We propose that stratification of GCs into EBV negative, EBVlow and EBVhigh may provide indicator value for the use of IDO1 inhibitors as adjuvant therapy against hiEBVaGCs.
EFFECT OF BETA-BLOCKADE ON POST-BURN FAT TISSUE SIGNALING
Finnerty CC1,2, Ali A1,2, Andersen C1,2, Kudlicki AS1,2, Tompkins RG3, Herndon DN1,2 1University of Texas medical Branch, Galveston TX, TX, USA; 2Shriners Hospitals for Children, Galveston, TX, USA; 3Massachussets General Hospital, Boston, MA, USA
OBJECTIVES/SPECIFIC AIMS: Severe burn injury induces massive inflammatory and lipolytic responses within the adipose tissue. Beta-blockade reduces peripheral lipolysis and inflammation in the severely burned. The effects of beta-blockade at the molecular level in adipose tissue, however, are unknown. METHODS/STUDY POPULATION: Adipose tissue samples from 39 burn patients, 21 control and 18 propranolol-treated, were harvested from the time of admission until 1 year post-burn. Propranolol was administered for up to 1 year post injury to decrease heart rate by ∼20%. Adipose gene expression was analyzed using the Affymetrix U133 plus 2.0 GeneChip™. An ANOVA was used to model the effects of treatment group, time post injury, burn size, sex, and age on gene expression. Clinical data detailing patient course were collected prospectively. RESULTS/ANTICIPATED RESULTS: Propranolol-mediated alterations in adipose tissue mRNA abundance were observed in 1,039 genes (p < 0.05 and false discovery rate <0.3, control vs. propranolol). Gene expression models were compared days 6 and 365 days post burn. The most significant effects of propranolol on gene expression correlated with a reduction in inflammatory response, increase in cellular proliferation, and decreases in apoptotic signaling, triacyglycerol biosynthesis, and beta-2 adrenergic receptor signaling. DISCUSSION/SIGNIFICANCE OF IMPACT: We identified alterations in adipose tissue gene expression induced by long-term propranolol treatment that suggest for the first time, that cellular proliferation, reduction of inflammation, and decreased cell death may contribute to the maintenance of peripheral adipose tissue seen with long-term propranolol administration.
ENDOTHELIAL CELL AND PERICYTE INTERACTIONS IN A SPHEROID MODEL OF ANGIOGENESIS
Chang WG1, Andrejecsk JW1, Kluger MS1, Saltzman WM1, Pober JS1 1Yale University, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Endothelial cell (EC) interactions with mural cells called pericytes (PCs) are critical to development, stabilization, and function of the microvasculature. Abnormal EC-PC interactions have been implicated in the development of diabetic microangiopathy and cancer. We wished to develop a model for analyzing these interactions using cultured human cells. METHODS/STUDY POPULATION: To examine EC-PC interactions, we adapted a cellular spheroid model using Bcl-2-transduced human umbilical vein ECs (Bcl-2-EC) to extend cell survival in 3-D collagen gels. RESULTS/ANTICIPATED RESULTS: Both culturing human placental PCs across a transwell from or addition of PC-conditioned media to Bcl-2-EC spheroids increase VEGF-A-induced sprouting. The majority of this effect is mediated by PC-derived hepatocyte growth factor (HGF), the production of which is enhanced by EC-conditioned medium and blocked by a PDGF receptor (R) tyrosine kinase inhibitor but not by siRNA knock down of PDGFR-β. In mixed Bcl-2-EC-PC spheroids, PCs closely invest EC sprouts but fail to stimulate Bcl-2-EC sprouting. PC coverage of Bcl-2-EC sprouts is reduced by the same PDGFR kinase inhibitor or by siRNA knockdown of PDGFR-β; reduction of the investment of sprouts by PCs restores PC-enhanced Bcl-2-EC sprouting. DISCUSSION/SIGNIFICANCE OF IMPACT: We have observed bidirectional EC-PC interactions using a cellular spheroid model. EC-conditioned medium induces PCs to secrete HGF which stimulates EC sprouting in a paracrine manner. However, EC sprouts recruit PCs and PC investment of EC sprouts blocks the stimulatory effect that PCs exert on sprouting. EC-derived signals that stimulate HGF secretion or PC coverage of EC sprouts may differ.
ENRICHMENT OF LUNG MICROBIOME WITH SUPRAGLOTIC MICROBES IS A RISK FACTOR FOR INCREASED PULMONARY INFLAMMATION
Segal LN1, Alekseyenko A1, Wu B1, Rom W1, Blaser M1, Weiden M1 1NYU School of Medicine, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: Micro-aspiration is a risk factor for COPD exacerbation while antibiotics reduce their frequency and severity. We hypothesize that enrichment of the lower airway microbiome with supraglotic microbes would predict airway inflammation in early COPD. METHODS/STUDY POPULATION: We studied 9 normal volunteers and 20 asymptomatic smokers with emphysema observed on screening CT (GOLD 0–1). We identified two well-defined lung microbiome (pneumotype) that we use to compare with their lung inflammatory cells and cytokines. Nasal bronchoscopy retrieved supraglotic and lower airway samples. Bronchoalveolar Lavage (BAL) cells and BAL cytokines defined pulmonary inflammation. Quantitative PCR measured 16S rRNA gene concentration and 454 sequence defined the microbiome. Sequences were analyzed with unsupervised hierarchical clustering, UniFrac and principle component analysis. RESULTS/ANTICIPATED RESULTS: As compared with Normal Volunteers, Emphysema cases had more neutrophils and inflammatory cytokines in their BAL. Cases and controls had the highest 16S rRNA concentration in supraglotic samples, BAL had intermediate bacterial 16S rRNA and saline had the lowest. There was no significant difference in the microbiome of normal controls and cases with early emphysema. Pneumotype 1 was not significantly different from the saline background in rDNA concentration or microbial community. It occurred in 5/9 controls and 12/20 cases. Compared to Pneumotype 1, Pneumotype 2 has higher rDNA concentration, high relative abundance of supraglotic flora such as Prevotella and Veillonella, higher BAL neutrophils and higher BAL lymphocytes. DISCUSSION/SIGNIFICANCE OF IMPACT: In both cohorts, a pneumotype with high relative abundance of supraglotic bacteria, such as Prevotella and Veillonella, is associated with subclinical inflammation.
EPIGALLOCATECHIN GALLATE (EGCG) ENHANCES IONIZING RADIATION INDUCED CELL DEATH IN CHONDROSARCOMA
Tang J1, He Z1, Zhang H1, Geller DS1, Guha C1, Sun HB1 1Albert Einstein College of Medicine/Montefiore Medical Center, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: Current effective treatment of chondrosarcoma is limited. Treatment that induce chondrosarcoma cells apoptosis and/or sensitize cells to ionizing radiation would be a promising option. While green tea derived EGCG has shown anticancer effect, it is not known whether EGCG exert a combinatorial effect with ionizing radiation on chondrosarcoma. This study is sought to test whether EGCG sensitizes chondrosarcoma to radiation and to identify the potential underlying mechanisms. METHODS/STUDY POPULATION: Human chondrosarcoma cell line SW1353, chondrocyte cell line C28/I2, and human primary chondrocytes were treated with (50, 100, 200, 400 uM) EGCG for 1–5 days with or without 137Cs γ-radiation. The harvested cells were subjected to MTT assay, cell counting, colony formation assay or qRT-PCR for expression of Cited2, a transcription regulator, and Bax/Bcl-2 (apoptosis). RESULTS/ANTICIPATED RESULTS: Treatment of EGCG showed toxicity to SW1353 cells in a dose and time dependent manner with ED50 at 200 uM at day 2. Treatment of SW1353 cells with 100 uM or 200 uM EGCG suppressed cell viability and colony formation by radiation (5 Gy, 10 Gy) in an additive fashion. Furthermore, EGCG significantly up-regulated Cited2 after 3 hr and Bax/Bcl-2 ratio after 24 hrs, which was associated with increased cell death of SW1353 chondrosarcoma. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results demonstrated that 1.) EGCG has an additive effect with ionizing radiation to induce cell death in chondrosarcoma. 2.) The anticancer effect may be regulated by Bax/Bcl-2 and Cited2. Of interest, the observation that Cited2 is modulated by EGCG suggested a novel anticancer role of Cited2 in chondrosarcoma. Further study on the role of EGCG in enhancing radiation therapy for chondrosarcoma is warranted in order to enlarge our current repertoire of treatments for this rare disease.
EPIGENOMIC DYSREGULATION IN PRE-ECLAMPSIA
Suzuki M1,3, Heo H1, Gebb JS1, Reznik SE2,1, Einstein MH1, Einstein FH1, Greally JM1 1Albert Einstein College of Medicine, Bronx, NY, USA; 2College of Pharmacy and Allied Health Professions, St. John's University, Queens, NY, USA; 3Clinical Research Training Program, Albert Einstein College of Medicine, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: Pre-eclampsia (PE) is a multisystemic pregnancy-associated disorder characterized by maternal hypertension and proteinuria after 20 weeks of gestation. This pilot study investigated the hypothesis that alteration of DNA methylation in the placenta can be found in women who have PE. METHODS/STUDY POPULATION: After IRB approval and informed consent, we collected the amnion of placentas of women with PE (n = 4) and with normal gestation (n = 6). We focused on the amnion rather than other placental components in order to reduce the variability between the samples that may potentially reflect cell composition rather than true epigenetic differences occurring at the cellular level. We analyzed the changes in DNA methylation with genome-wide profiling (HELP-tagging) and targeted high resolution profiling (targeted MethylC-seq). RNA expression profiles were analyzed using RNA-seq. RESULTS/ANTICIPATED RESULTS: Thirty five statistically significantly differentially methylated loci from genome-wide analysis (p < 0.001 and 5% FDR) were identified. The targeted high resolution profiling showed promoter hypomethylation of soluble FLT1 (sFLT1) isoforms in the amnion in PE. RNA-seq analysis showed 32 differentially expressed genes (fold difference > 1.5). DISCUSSION/SIGNIFICANCE OF IMPACT: We observed DNA methylation and RNA expression status alterations in PE and might reflect the natural responses to PE or be causally related to PE. An expanded prospective study may provide further insights in the pathogenesis of PE and may lead to novel biomarkers for early identification and diagnosis.
EVALUATION OF THE CD4 T CELL RESPONSE AGAINST INFLUENZA FOLLOWING HEMATOPOIETIC CELL TRANSPLANTATION
Knorr D1, Karras N1, Weeres M1, Verneris MR1 1University of Minnesota, Minneapolis, MN, USA
OBJECTIVES/SPECIFIC AIMS: Bone marrow transplant is a curative treatment for patients with leukemia. However, high doses of chemotherapy and radiation given to abolish resident cancer cells and allow successful engraftment of donor hematopoietic stem cells (HSCs) leads to profound immunodeficiency. Patients remain susceptible to infections for months to years following transplant. Although vaccination can provide some protection against viruses such as influenza, there is limited mechanistic evidence on the immune cell subsets directly involved. In order to better understand the response to vaccination in the transplant setting, a more specific analysis of the generation of influenza specific CD4+ T helper cells is needed. METHODS/STUDY POPULATION: These studies utilize T cell tetramer reagents and multi-color flow cytometry to analyze the cytokine responses of influenza specific T cells in HCT recipients. RESULTS/ANTICIPATED RESULTS: Contrary to our initial study hypothesis, administration of an additional dose of influenza vaccine did not improve the T cell or B cell responses in either group. Therefore, we hypothesize that at time of vaccination, the frequency of hemagglutinin peptide (HAp):DR4-specific CD4+ T cells following HCT will predict vaccine response. We also predict that recipients of BM donors mount a more robust response due to pre-existing HAp:DR4-specific memory T cells. DISCUSSION/SIGNIFICANCE OF IMPACT: Studying the antigen-specific response to influenza vaccination is critical to understand the mechanisms driving current clinical care. Direct correlation of the post-transplant influenza-specific T cell repertoire to vaccine responsiveness will allow us to adapt alternative strategies to treat patients post-HCT and lessen the morbidity and mortality caused by influenza.
EXCESSIVE AND ECTOPIC VASCULAR SMOOTH MUSCLE CELLS IN PULMONARY HYPERTENSION
Greif D1, Sheikh A1 1Yale University School of Medicine, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: The overall goal of this project is to elucidate the origin(s) of excess pulmonary artery smooth muslce cells in mouse models of pulmonary hypertension and the mechanisms underlying their recruitment, proliferation and differentiation. The project has two specific aims: 1) establish a timeline of molecular marker expression and cellular events in the left pulmonary artery and left lung arterioles and capillaries that define steps in the pathogenesis of pulmonary hypertension; 2) determine the cellular origin and clonal relationship of cells that comprise the increased pulmonary artery smooth muscle cell burden in mouse models of pulmonary hypertension. METHODS/STUDY POPULATION: To this end, histochemical, clonal and genetic analysis of transgenic mice will be integrated with studies of gene expression, cell migration and proliferation in human pulmonary artery cells. RESULTS/ANTICIPATED RESULTS: We hypothesize that the accumulation of pulmonary artery smooth muscle cells in pulmonary hypertension occurs through smooth muscle cell dedifferentiation, proliferation and migration and endothelial cell transdifferentiation. DISCUSSION/SIGNIFICANCE OF IMPACT: This work promises to reveal novel findings regarding the origin of excess smooth muscle cells in pulmonary hypertension and the cellular events underlying their accumulation. Futhermore, novel therapeutic strategies to combat this devastating disease will be suggeted.
EXPRESSION OF CARDIOLIPIN BIOSYNTHESIS AND REMODELING ENZYMES IN ADULT HEART FAILURE
Grudis JE1, Chatfield KC2, Sparagna GC4, Hijmans J3, Sobus RD3, Miyamoto SD2, Stauffer BL3,4 1University of Colorado School of Medicine, Aurora, CO, USA; 2Department of Pediatric Cardiology, Children's Hospital Colorado, Aurora, CO, USA; 3Department of Medicine, University of Colorado Denver, Aurora, CO, USA; 4Department of Integrative Physiology, University of Colorado Boulder, Boulder, CO, USA
OBJECTIVES/SPECIFIC AIMS: Cardiolipin (CL) is a unique phospholipid that is an essential component of the inner-mitochondrial membrane critical for normal energy metabolism. Biosynthesis of CL occurs via an enzymatic pathway or through remodeling of existing CLs. The content of total CL and (18:2) 4CL (tetralinoleic form, normally predominant in the heart) are lower in ventricular tissue from adults with heart failure secondary to idiopathic dilated cardiomyopathy (IDC). The aim of this study was to determine the expression levels of CL biosynthetic and remodeling enzymes associated with CL content changes in IDC. METHODS/STUDY POPULATION: mRNA was isolated from the left ventricle (LV) of adult IDC patients at transplant (n = 27; mean age = 51+/-14) and non-failing control LV from donor hearts (n = 15, mean age = 43+/-8). RT-PCR was used to measure expression of CL biosynthesis and remodeling enzymes. RESULTS/ANTICIPATED RESULTS: Expression of biosynthetic enzymes, CDP diacylglycerol synthase 2 and phosphatidylglycerolphosphate synthase, were both 33% lower in IDC LV (p < 0.05). Remodeling enzymes, monolyso-CL acyltransferase and tafazzin, were also down-regulated (66% and 43% respectively, p < 0.01) in IDC LV compared to controls. DISCUSSION/SIGNIFICANCE OF IMPACT: These results demonstrate that biosynthetic and remodeling CL abnormalities are present in failing IDC hearts and may contribute to mitochondrial CL abnormalities in heart failure. Supported by NIH/NCATS Colorado CTSI Grant Number UL1 TR000154.
FGF23 IN THE DEVELOPMENT OF LEFT VENTRICULAR HYPERTROPHY IN CHRONIC KIDNEY DISEASE
Ichikawa S1, Unsicker TR1, Alkhouli M1, Gray AK1 1Indiana University, Indianapolis, IN, USA
OBJECTIVES/SPECIFIC AIMS: Chronic kidney disease (CKD) leads to increased left ventricular hypertrophy (LVH). In early stages of CKD, the phosphaturic hormone, fibroblast growth factor 23 (FGF23), begins to rise to maintain blood phosphorus levels in the normal range. However, in advanced stages, hyperphosphatemia ensues despite markedly elevated FGF23. Both hyperphosphatemia and elevated FGF23 levels are associated with LVH in CKD. The objective is to dissect the relative contributions of elevated phosphorus and FGF23 to LVH. METHODS/STUDY POPULATION: The Jck mice, a murine model of CKD, were bred with Galnt3 knockout mice, which have the unique ability to genetically reduce the amount of circulating intact Fgf23. All mice were fed high phosphate diet from weaning and sacrificed at 12 weeks of age. Serums were analyzed for phosphorus, calcium, alkaline phosphatase, creatinine, parathyroid hormone (PTH), and FGF23. Heart mass index (heart weight/body weight) was determined from the harvested hearts. RESULTS/ANTICIPATED RESULTS: Intact Fgf23 levels in Jck and Jck/Galnt3 double mutant mice were 2 and 12 fold higher than that in normal controls. Both Jck and double mutant mice had higher serum phosphorus levels; however, the double mutants were significantly more hyperphosphatemic than the Jck mice. The Jck mice had the highest heart mass index, while the index for the double mutants were intermediate between the normal and Jck mice. DISCUSSION/SIGNIFICANCE OF IMPACT: Increased heart mass index in the Jck and double mutant mice indicates the presence of LVH. The Jck mice, which were less hyperphosphatemic with higher Fgf23, had higher heart mass than more hyperphosphatemic double mutant mice, which had lower Fgf23 levels. This observation suggests that elevated Fgf23, rather than hyperphosphatemia, likely causes LVH in CKD.
GENE EXPRESSION PROFILE AND PLASMA CYTOKINE RESPONSE TO INTRAVENOUS (IV) ALCOHOL IN HUMANS
Coe MA1, Tapocik JD1, Sommer WH1, Solomon M1, Flanigan M1, Singley E1, Leggio L1, Heilig M1, Ramchandani VA1 1NIAAA/NIH, Bethesda, MD, USA
OBJECTIVES/SPECIFIC AIMS: Alcohol can activate pro-inflammatory cascades in multiple tissues, and alterations in peripheral cytokine signaling pathway gene expression have been shown in alcoholics. However, peripheral cytokine response to acute IV alcohol has not been studied. Therefore, we examined changes in peripheral gene expression profiling (GEP) and cytokine levels in response to IV alcohol in healthy social drinkers. METHODS/STUDY POPULATION: 48 social drinkers underwent 2 single-blind sessions and received, on separate days, IV infusions of saline (placebo) or 6%v/v alcohol for 3 hrs to achieve and clamp a target breath concentration of 50 mg%. Blood samples were collected at baseline and end of the infusion for GEP and cytokine levels. RESULTS/ANTICIPATED RESULTS: GEP (n = 9) identified 105 genes differently regulated after IV alcohol infusion (45 upregulated, 60 downregulated). Bioinformatic analysis determined an over-representation of inflammatory pathways post-alcohol infusion (e.g. LPS/IL-1 Mediated Inhibition of RXR Function, IL-6 Signaling). Cytokine levels (n = 28) identified a time by age effect and trend for time by treatment by age effect for IL1B and a gender by time interaction and a time by age interaction for TNF-alpha. DISCUSSION/SIGNIFICANCE OF IMPACT: This study demonstrated that inflammatory protein-level changes occur in healthy social drinkers after acute alcohol infusion, and more specifically two proteins critically involved in the pro-inflammatory cascade: IL1B and TNF-alpha. As altered cytokine signaling is seen in alcoholics, determining the development of abnormalities in these pathways may be helpful for detection and treatment of alcohol-related medical problems. This pilot study suggests that changes in cytokine-signaling proteins may serve as an early biomarker of alcohol-related disorders.
GENOMIC VARIATION AND PROCEDURAL ANXIETY
Ersig A1, Hanrahan K1, Standley J2, Kleiber C1, McCarthy A1, Murray J2 1University of Iowa College of Nursing, Iowa City, IA, USA; 2University of Iowa Carver College of Medicine, Iowa City, IA, USA
OBJECTIVES/SPECIFIC AIMS: Despite significant gains in knowledge of the human genome over the past decade, little is known about the association of genomic variants with outcomes in children. In a two-phase intervention study of responses to IV insertion, DNA and measures of child anxiety were gathered from children and biological parents. This analysis examines association of single nucleotide polymorphisms (SNPs) and child anxiety. METHODS/STUDY POPULATION: The study enrolled 1124 children age 4–10, who had IVs placed. Outcome variables were child and parent ratings of procedural and trait anxiety and measures of child distress. DNA was extracted from whole blood, buccal swab, or saliva samples. Candidate genes were selected based on involvement in pathways of interest; SNPs were selected based on prior association with child anxiety and distress. RESULTS/ANTICIPATED RESULTS: Genes included COMT, CRH, CRHR1, EDN1, EDNRA, GABRA2, GRM3, MC1R, and TAC1. Six SNPs were associated with high anxiety; 2 were also associated with high pain. SNPs associated with high anxiety are in genes associated with the HPA axis, peripheral nociception and central nervous system regulation. DISCUSSION/SIGNIFICANCE OF IMPACT: Future analyses are planned to explore pathways involved and determine gene-gene or gene-environment interactions. TDT analysis is planned on DNA obtained from biologically related trios.
GENOMICS AND NETWORK ANALYSIS OF RTPA LYSIS IN A QUANTITATIVE CLOT MODEL
Chan Y1, Hao K1, Kasarskis A1, Richardson L1, Wang R1, Shaw G2, Meunier J2, Pancioli A2, Tuhrim S1, Schadt E1 1Mount Sinai School of Medicine, New York City, NY, USA; 2University of Cincinnati, Cincinnati, OH, USA
OBJECTIVES/SPECIFIC AIMS: 1. Generate Genome Wide Association (GWAS) and Exome chip data for 40 healthy volunteers to investigate the association between genotype and recombinant tissue plasminogen activator (rtPA) efficacy using a validated, quantitative ex-vivo assay 2. Generate peptidomic and proteomic characterizations of pooled rtPA-responsive and non-responsive individuals to identify candidate proteins and peptides associated with rtPA efficacy using quantitative proteomics and targeted protein analysis. 3. Conduct an exploratory integrative analysis of DNA variation, protein and peptide variation, clinical data, and quantitative measures of rtPA activity to identify constellations of analytes that predict outcome and may have potential as predictive biomarkers of rtPA efficacy. METHODS/STUDY POPULATION: Rater-blinded, prospective cohort genomics/proteomics association study of 40 healthy volunteers. 1. Use a clot visual microscopy technique; fractional clot-width loss will be as an assay to classify donors. 2. Use HumanOmniExpressExome DNA Analysis BeadChip to generate genotype data. 3. Conduct stable isotope labeling based quantitative proteomic experiment. 4. Perform integrated analysis of rtPA response, DNA variants and serum protein markers. RESULTS/ANTICIPATED RESULTS: Our translational, hypothesis-generating study is an essential step towards clinical validation of pharmacogenomic (PGx)/ pharmacoproteomic (PPx) findings from basic science and pilot human studies. The results will help inform the design of future large prospective trials. DISCUSSION/SIGNIFICANCE OF IMPACT: Our goal is to enable physicians to make more informed decisions in risk stratifying thrombolytic candidate patients, determine the best reperfusion strategy, and thereby provide optimal treatment for each individual patient.
GLUTATHIONE REDOX STATUS IN CALD PATIENTS FOLLOWING N-ACETYLCYSTEINE ADJUVANT THERAPY
Radtke KK1, Coles LD1, Mishra U1, Cloyd JC1 1University of Minnesota, Minneapolis, MN, USA
OBJECTIVES/SPECIFIC AIMS: Cerebral adrenoleukodystrophy (cALD) is a rare, X-linked disorder associated with increased oxidative stress and progressive neurological damage. The addition of N-acetylcysteine (NAC) therapy to hematopoietic stem cell transplantation (HSCT) in advanced cALD patients has resulted in significant improvements in overall survival. NAC increases intracellular glutathione, an endogenous antioxidant. The ratio of reduced (GSH) to oxidized (GSSG) glutathione, also known as the GSH redox ratio, in whole blood is used as an indicator of oxidative stress. The objective of this study was to measure the effect of NAC intravenous therapy on the GSH redox ratio in whole blood of cALD patients who are undergoing HSCT. METHODS/STUDY POPULATION: cALD boys were administered 70 mg/kg NAC intravenously every 6 hours for ∼100 days following HSCT (Day 0). Two mL of whole blood was collected prior to and at various times 15–300 min post-NAC infusion on Day+7 and Day+21 of therapy. Relative amounts of NAC, GSH, and their oxidized forms were determined by HPLC-Mass Spectrometry. Redox status, calculated as the GSH/GSSG ratio, was determined at each time point. RESULTS/ANTICIPATED RESULTS: Pre-infusion NAC concentrations were below the lower limit of detection. NAC and GSH increased after NAC infusion with the maximum concentrations observed at 15 min post-end of infusion. GSH/GSSH also increased following NAC infusion and was highest at 15 min post-infusion, gradually declining thereafter. DISCUSSION/SIGNIFICANCE OF IMPACT: These results demonstrate that one effect of NAC is to transiently modulate GSH/GSSG ratios in whole blood which provides a greater understanding of how NAC exerts its antioxidant effects. GSH redox ratio may be a key parameter to optimize and monitor NAC therapy in cALD as well as other diseases associated with oxidative stress.
GOLD AND HYDROXYAPATITE NANO-GRAFTS FOR ANTERIOR CRUCIATE LIGAMENT RECONSTRUCTION
Smith S1, Grant S1, White R1 1University of Missouri, Columbia, MO, USA
OBJECTIVES/SPECIFIC AIMS: The objective of this study is to design anterior cruciate ligament (ACL) grafts from the extracellular matrix (ECM) of allograft and xenograft tissue, gold nanoparticles (AuNP) and hydroxyapatite nanoparticles (HANP). There is a clinical need for improved graft materials due to frequent failure caused by a lack of remodeling. AuNP have been shown to enhance cellular remodeling as well as cellular attachment and proliferation. HANP are proposed to enhance bone cell attachment and mineralization. This study aims to design, synthesize and characterize these novel scaffolds. METHODS/STUDY POPULATION: Porcine diaphragms were harvested and decellularized in tributyl phophate in Tris buffer solution. Nanoparticles were crosslinked to the tissue using a zero-length carbodiimide crosslinker then sterilized in a paracetic acid sodium chloride solution. The scaffold was characterized using scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and fibroblast cell assays. RESULTS/ANTICIPATED RESULTS: SEM results confirmed successful crosslinking of nanomaterials to the scaffolds. DSC results confirmed no significant thermal damage when crosslinking nanoparticles and sterilizing samples. Cell culture assays showed biocompatibility of scaffolds at a range of concentrations of nanoparticles with improved biocompatibility using <40 nm plate-like HANP over <200 nm spherical HANP. DISCUSSION/SIGNIFICANCE OF IMPACT: Using current decelluarization, crosslinking and sterilizing techniques, gold and hydroxyapatite nano-grafts have been synthesized and characterized. Neither AuNP or HANP showed toxicity to cells in fibroblast cell assays indicating biocompatibility. The proposed graft shows promise to be an improved ACL graft over current materials.
HIV-1 THERAPY BY A COMBINATION OF BROADLY NEUTRALIZING ANTIBODIES IN HUMANIZED MICE
Klein F1, Halper-Stromberg A1, Horwitz JA1, Gruell H1, Scheid JF1, Bournazos S1, Mouquet H1, Linda SA1, Diskin R2, Abadir A1, Zang T3, Dorner M1, Billerbeck E1, Labitt RN1, Gaebler C1, Marcovecchio PM2, Incesu R1, Eisenreich TR1, Bieniasz PD3, Seaman MS4, Bjorkman PJ2, Ravetch JV1, Ploss A1, Nussenzweig MC1 1The Rockefeller University, New York, NY, USA; 2Division of Biology, California Institute of Technology, Pasadena, CA, USA; 3Aaron Diamond AIDS Research Center, Laboratory of Retrovirology, New York, NY, USA; 4Beth Israel Deaconess Medical Center, Boston, MA, USA
OBJECTIVES/SPECIFIC AIMS: Human antibodies to human immunodeficiency virus-1 (HIV-1) can neutralize a broad range of viral isolates in vitro and protect non-human primates against infection. Previous work showed that antibodies exert selective pressure on the virus but escape variants emerge within a short period of time. However, these experiments were performed before the recent discovery of more potent anti-HIV-1 antibodies and their improvement by structure-based design. METHODS/STUDY POPULATION: Here we re-examine passive antibody transfer as a therapeutic modality in HIV-1-infected humanized mice. RESULTS/ANTICIPATED RESULTS: Although HIV-1 can escape from antibody monotherapy, combinations of broadly neutralizing antibodies can effectively control HIV-1 infection and suppress viral load to levels below detection. Moreover, in contrast to antiretroviral therapy, the longer half-life of antibodies led to control of viremia for an average of 60 days after cessation of therapy. DISCUSSION/SIGNIFICANCE OF IMPACT: Thus, combinations of potent monoclonal antibodies can effectively control HIV-1 replication in humanized mice, and should be re-examined as a therapeutic modality in HIV-1-infected individuals.
HOMOZYGOSITY MAPPING AND WHOLE EXOME SEQUENCING (WES) FOR THE IDENTIFICATION OF AUTISM AND INTELLECTUAL DISABILITY (ID) RISK GENES
Ercan-Sencicek AG1, Jambi S4, Yuksel Z2, Herguner S3, Oztop D5, Fidan T2, Artan S2, Gunel M1, State MW1 1Yale University, New Haven, CT, USA; 2Osmangazi Univ, Eskisehir, Turkey; 3Necmettin Erbakan Univ, Konya, Turkey; 4Alhada Armed Hospitals, Taif, United Arab Emirates; 5Erciyes Univ, Kayseri, Turkey
OBJECTIVES/SPECIFIC AIMS: The substantial genetic and phenotypic heterogeneity of neurodevelopmental disorders has made the identification of candidate genes very challenging. Studying the consanguineous families that have been used to seek regions of homozygosity in affected individuals is a powerful approach to identifying mechanisms of disease. Combination of genotyping and WES have been demonstrated as an effective approach to identify rare homozygous mutations within the coding portion of the genome and to simultaneously identify homozygous stretches of identity by descent from the genotyping data. METHODS/STUDY POPULATION: Over the last 12 months, an effort for the recruitment of families with autism and ID for gene mapping from Turkey and Saudi Arabia resulted in the identification of 36 consanguineous pedigrees in which the proband carries this diagnosis. Eighteen of these are simplex pedigrees consisting of 72% 1st cousin pairings and 18 are multiplex composed of 78% 1st, 17% 2nd, and 5% 3rd cousin matings. mutations in consanguineous pedigrees will be searched in both unrelated consanguineous and outbred families to evaluate the convergence of risk genes. RESULTS/ANTICIPATED RESULTS: Blood derived DNA from parents and affected children in each pedigree will be genotyped to pursue homozygosity mapping and CNV analysis followed by WES of affected individuals to identify rare variants of large effect. Finally, the identified homozygous loss of function. DISCUSSION/SIGNIFICANCE OF IMPACT: The discovery of additional rare recessive, penetrant ASD and ID mutations in consanguineous families will help the understanding the molecular mechanisms of autism and ID in outbred populations.
HUMAN CD26BRIGHT TH17 CELLS ERADICATE LARGE TUMORS
Paulos CM1, Huff LW1, Kundimi S1, Goodyear M1, Nelson MH1 1MUSC, Charleston, SC, USA
OBJECTIVES/SPECIFIC AIMS: Human CD4+ T cells can differentiate into multiple effector subsets, but their role in antitumor immunity remains incompletely elucidated. We seek to determine how distinct human CD4+ T cell subsets impact antitumor immunity in mice with large human malignancies. METHODS/STUDY POPULATION: We sorted Th1 (CXCR3+), Th2 (CCR4+), or Th17 (CCR6+ or CD26hi) subsets from bulk human CD4+ T cells, stimulated them with CD3/CD28 or CD3/ICOS beads and engineered them with a chimeric antigen receptor (CAR) that recognizes mesothelioma tumors. RESULTS/ANTICIPATED RESULTS: We found that ICOS costimulation not only augmented human Th17 cells' function, but also enhanced the function of human Th1 and Th2 cells superior to CD28 costimulation. Moreover, CD26hi Th17 cells were more polyfunctional than the other T cells subsets, as indicated by increased secretion of IL-17, IFN-γ, IL-22 and TNF-α. Of clinical importance, CD26hi Th17 cells mediated superior regression of large human mesothelioma in mice compared to CXCR3+Th1, CCR4+Th2 or CCR6+Th17 cells. The therapeutic effectiveness of CD26hi Th17 cells was associated with enhanced functionality and persistence in vivo. DISCUSSION/SIGNIFICANCE OF IMPACT: Taken together, these data indicate that the appropriate selection of effector CD4+ T cell subsets is critical for successful tumor eradication. These findings will help guide next generation clinical trials for patients with advanced malignancies.
IDENTIFICATION OF MIRNA TARGETS IN BREAST TUMORS USING BIOINFORMATIC ANALYSIS OF COORDINATED MIRNA-MRNA PATIENT DATA AND AGO-PAR-CLIP
Farazi TA1, Brown M1, Mihailovic A1, Tuschl T1, ten Hoeve JJ2, Wessels L2 1Rockefeller University, New York, NY, USA; 2Netherlands Cancer Institute, Amsterdam, Netherlands
OBJECTIVES/SPECIFIC AIMS: miRNAs are known to regulate genes critical for tumorigenesis. However, identifying their direct mRNA targets mediating their cellular functions remains a challenging task. It is mainly based on computational target prediction and testing of promising candidates, chosen by a priori biological pathway knowledge, in in vitro and in vivo assays. METHODS/STUDY POPULATION: To elucidate and prioritize miRNA-mRNA targets in breast cancer we adopted a dual approach: (1) a biochemical approach, AGO Photoreactive-Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation (PAR-CLIP), allowing genome-wide identification of AGO-bound transcripts in the breast cancer cell line MCF7, and a (2) bioinformatic analysis of a large (n = 161) well-characterized patient collection with coordinated miRNA-mRNA data along with clinical information, using sequencing-derived miRNA profiles. RESULTS/ANTICIPATED RESULTS: The AGO-PAR-CLIP approach resulted in a list of biochemically validated associated miRNA-mRNA targets with binding site resolution. For correlation analysis of coordinated miRNA-mRNA data from the patient collection, we derived abundance filters for miRNA and mRNA data that strengthened (1) the positive correlations between intronic miRNAs and their host mRNAs, and (2) the negative correlations between miRNAs and their computationally predicted targets. DISCUSSION/SIGNIFICANCE OF IMPACT: Using this dual approach, we ranked miRNAs for their targeting activity, defined their associated pathways, and assessed whether in combination with their targets predicted patients that developed distant metastases.
IDENTIFICATION OF SMALL MOLECULE INHIBITORS OF JARID1B HISTONE DEMETHYLASE
Sayegh J1, Cao J1, Morales A1, Blair LP1, Zou MR1, Norcia M2, Hoyer D2, Merkel JS2, Yan Q1 1Department of Pathology, Yale University School of Medicine, New Haven, CT, USA; 2Yale Center for Molecular Discovery, Yale University, West Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: JARID1B (also known as KDM5B or PLU1) is a member of the JARID1 family of histone lysine demethylases responsible for the demethylation of trimethylated lysine 4 in histone H3 (H3K4me3), a mark for actively transcribed genes. JARID1B is over-expressed in several cancers, including breast cancer, prostate cancer and lung cancer. Furthermore, JARID1B expressing melanoma cells are associated with increased self-renewal character. Therefore, JARID1B represents an attractive target for cancer therapy. The aim of this study is to identify specific small molecule inhibitors of JARID1B. METHODS/STUDY POPULATION: First we characterized JARID1B using a homogeneous luminescence-based demethylase assay. We then conducted a high-throughput screen of over 15,000 small molecules to identify inhibitors of JARID1B demethylase. RESULTS/ANTICIPATED RESULTS: From this screen, we identified several known JmjC histone demethylase inhibitors and a novel inhibitor which inhibits JARID1B with an IC50 of 3 μM in vitro. Consistent with this, treatment of several cell lines with this novel inhibitor leads to global increase of H3K4me3 level. Furthermore, this compound inhibited proliferation of two human breast cancer cell lines, but not immortalized mammary epithelial cells. DISCUSSION/SIGNIFICANCE OF IMPACT: These results suggest that this novel small molecule inhibitor is a lead compound that can be further optimized for cancer therapy.
IDENTIFYING AND CHARACTERIZING “C-MYC-ADDICTED” NON-SMALL CELL LUNG CANCERS (NSCLC)
Dospoy P1, Chen P1, Soucek L2, DeBerardinis R1, Minna J1 1UT Southwestern Medical Center, Dallas, TX, USA; 2Vall D'Hebron Institute of Oncology, Barcelona, Spain
OBJECTIVES/SPECIFIC AIMS: MYC is overexpressed and in some cases amplified in lung cancer. However, it is unknown whether some lung cancers are “addicted” to MYC expression for their survival and tumorigenic phenotype. Our goal is to identify lung cancers that are addicted to MYC, determine their oncogenotype and molecular properties, and “acquired vulnerabilities” that would provide both novel therapeutic targets and enrollment biomarkers to identify patients with MYC addicted tumors for new targeted therapy. METHODS/STUDY POPULATION: We studied a large panel of clinically and molecularly annotated NSCLC lines for MYC mRNA and protein expression, DNA copy number, and metabolic profiles. Functional tests were performed on 12 NSCLCs with dominant negative OMOMYC and effects monitored by testing for expression of MYC downstream targets and by colony forming efficiency assays (CFE). RESULTS/ANTICIPATED RESULTS: In all cases, exogenous OMOMYC expression led to down regulation of the target genes. 4/12 NSCLCs were dramatically sensitive to OMOMYC (with >100 fold reduction in CFE) compared to 7/12 totally resistant and one with an intermediate response. The sensitive NSCLCs were KRAS wild-type (one EGFR mutant) and exhibited epithelial morphology, while 5/7 resistant NSCLCs were KRAS mutant, most of which exhibited mesenchymal morphology. Surprisingly, there was no correlation with MYC mRNA and protein expression or DNA copy number. DISCUSSION/SIGNIFICANCE OF IMPACT: We conclude: there is a subset of NSCLCs that demonstrates dramatic growth inhibition by OMOMYC which we are classifying as “MYC addicted” which are wild-type for KRAS and epithelial. Using the MYC “addicted” vs. non- addicted NSCLC panel, we are testing for potential downstream MYC therapeutic targets.
IMAGING NICOTINIC ACETYLCHOLINE RECEPTORS IN THE PSYCHOSIS PRODROME: STUDY PROPOSAL
Carlson J1, D'Souza D1, Woods S1, Mayes L1, Picciotto M1 1Yale, Hamden, CT, USA
OBJECTIVES/SPECIFIC AIMS: Up to 88% of individuals with schizophrenia (SZ) smoke tobacco versus approximately 22% in the general population. Resumption of tobacco smoking after overnight abstinence led to improved performance on a VisouSpatial Working Memory (VSWM) task smokers with SZ, an enhancement not found in controls. Postmortem & in Vivo studies have found that smoking increases the nAChR subtype containing beta subunits (β2*nAChR) in a number of brain regions in smokers without mental illness. Smokers with SZ do not demonstrate such upregulation [2,3]. Whether abnormalities in β2*nAChR availability are a consequence of developing SZ or are present at the onset of the illness is not known. The proposal aims to test the hypotheses that: 1)There are reductions in high affinity β2*nAChR availability in prodromal smokers which falls between that of smokers with SZ & non-psychiatric smokers. 2)Abstinence from nicotine impairs VSWM performance in prodromal smokers which is restored by smoking. METHODS/STUDY POPULATION: In 15 Prodromal subjects VSWM, symptom & smoking measures will be assessed at 1)during ad libitum smoking; 2) 24–48hrs post-smoking cessation: during nicotine withdrawal; 3) Day 6: after sustained short-term nicotine abstinence; 4) if the subject resumes smoking. β2*nAChR availability will be acquired by SPECT. Matched SZ & non-psychiatric smokers will be used for comparison. Cognitive data, symptom data and smoking data will be correlated with regional β2*nAChR availability. Refs: 1) Sacco K,et al. Effects cig smoking on spatial WM & attentional deficits in schiz. Ar gel psy. 2005; 62:649–659 2) Picciotto M,et al. Nicotinic receptors in the brain. Links between molecular biology & behavior. Neuropsypharm. 2000;22:451–465 3) D'Souza D,et al. Lower β2*nAChR availability in smokers with schiz. AmJPsyc. 2012; 169:326–334 RESULTS/ANTICIPATED RESULTS: . DISCUSSION/SIGNIFICANCE OF IMPACT.
IN VITRO INVESTIGATION OF OXIDATIVE STRESS IN NORMAL & MALIGNANT MURINE HEPATOCYTES
Reynolds L1, Mulik R1, Kim E1, Wen X1, Liao C1, Terada L1, Corbin I1 1University of Texas Southwestern Medical Center, Dallas, TX, USA
OBJECTIVES/SPECIFIC AIMS: Hepatocellular carcinoma (HCC) has a 3% 3-yr survival and chemotherapy is very toxic atop existing liver disease. Studying the inherent differences between HCC and normal liver tissue will help identify therapies that exploit unique weaknesses in HCC. The redox system maintains cellular homeostasis by generating and eliminating reactive oxygen species (ROS). In cancer, elevated oxidative stress promotes proliferation and blocks apoptosis, but extreme levels induce cell death. Our aim was to compare oxidative stress levels in normal (TIB73) and malignant (TIB75) mouse hepatocyte cells. METHODS/STUDY POPULATION: DCF and TBARS were used to investigate the levels of cellular ROS. Subcellular ROS levels were tracked using a targeted ratiometric fluorescent H2O2 sensor (HyPer). We measured major antioxidative enzyme protein levels by western blot. RESULTS/ANTICIPATED RESULTS: Fluorometric and FACS DCF readings indicated that ROS levels were 3–4 fold higher in TIB75 cells than TIB73 cells. HyPer was targeted to the ER, mitochondria, and cytoplasm. Fluorescent imaging revealed no difference in H2O2 production between TIB73 and TIB75 cells in the ER and mitochondria, but TIB73 cells did exhibit higher cytoplasmic H2O2 levels than TIB75 cells. TIB75 cells had ≈4 times higher levels of lipid peroxide species than TIB73 cells by TBARS. TIB75 cells also expressed significantly lower protein levels of catalase Superoxide dismutase-1 and glutathione peroxidase-4 levels were lower in TIB75, but were only approaching significance. DISCUSSION/SIGNIFICANCE OF IMPACT: These data show that HCC cells have greater oxidative stress and a reduced coping capacity than normal liver cells. This important biological difference may be exploited by therapies that increase ROS or weaken the antioxidant defenses in HCC to cause selective tumor toxicity and preserve the normal liver tissue.
INFLAMMATORY REGULATION IN PEDIATRIC CONGENITAL AORTIC VALVE STENOSIS: AN INITIAL REPORT FROM A CLINICAL PROTEOMICS STUDY
Angel PM1, Bichell D1, Ru Su Y1, Hinton R2, Mettler B1, Baldwin H1, Caprioli RM1 1Vanderbilt University Medical Center, Nashville, TN, USA; 2University of Cincinnati, Cincinnati, OH, USA
OBJECTIVES/SPECIFIC AIMS: Congenital aortic valve stenosis (CAVS) occurs in over 1.4% of U.S. births. CAVS is caused by fusion and progressive engorgement of aortic valve (AV) leaflets with disorganized extracellular matrix (ECM) narrowing the aortic valve orifice. The only treatment is surgical replacement of the AV. Our goal is to understand the cellular mechanisms of CAVS towards development of medical treatments that attenuate disease progression. METHODS/STUDY POPULATION: We are obtaining de-identified tissue from CAVS patients, donors with documented normal cardiac function (controls), and hemodynamically altered valves spanning neonate to adult. Each AV is analyzed in triplicate by multidimensional protein identification technology. Systems biology approaches are being used to develop molecular signatures of CAVS. RESULTS/ANTICIPATED RESULTS: Preliminary data from 18 patients produced a database of 3,105 proteins from normal AV and CAVS tissue. A total of 45 ECM proteins appear differentially regulated and have processes of 'response to external stimulus', 'wound healing', and 'acute inflammatory response'. Affected signaling pathways identified include the insulin, PPAR, Wnt, and TGFbeta signaling cascades. Interestingly, multiple pathways and networks link to inflammation and immune signaling in both normal and CAVS valves. DISCUSSION/SIGNIFICANCE OF IMPACT: Normal AV development involves inflammation processes that are altered in CAVS. Collective data suggests that processes of inflammation may be working in parallel with normal AV development as a protective mechanism, but are divergent in CAVS. Detailed investigations are in progress. These studies provide focused targets for development of animal models and offer potential opportunities for therapeutic intervention.
INHALED FORMOTEROL DIMINISHES INSULIN-INDUCED HYPOGLYCEMIA
Belfort De Aguiar R1, Naik S1, Szepietowska B1, Sherwin R1 1Yale University, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: The short-acting beta-2 agonist terbutaline has been reported to reduce nocturnal hypoglycemia in T1DM patients, findings that are consistent with our data showing that activation of beta-2 adrenergic receptors in the ventromedial hypothalamus stimulates counterregulatory responses in the rat. The current study examines whether inhalation of the long-acting beta-2 agonist formoterol, used clinically for asthma, reduces insulin-induced hypoglycemia in healthy volunteers (n = 7) and T1DM patients (n = 5). METHODS/STUDY POPULATION: Studies were conducted on two separate sessions: either with 48 mcg of formoterol or placebo through an aeroinhaler. Subsequently, a two step 2 mU/kg/min hyperinsulinemic hypoglycemic clamp was performed that was targeted to maintain glucose levels at 50–55 mg/dL for the last hr using a variable glucose infusion. RESULTS/ANTICIPATED RESULTS: Despite similar insulin and glucose concentrations, formoterol decreased the glucose infusion rate at the end of the clamp in healthy subjects by 27% (p = 0.01) and also tended to have an effect in T1DM subjects (by 43%, p = 0.056). No differences were seen, however, in counterregulatory hormone levels. A small increase was seen in heart rate during hypoglycemia in human subjects with formoterol, although rates never exceeded 100 bpm. DISCUSSION/SIGNIFICANCE OF IMPACT: Our data demonstrate that Inhaled formoterol can reduce insulin-induced stimulation of glucose metabolism during a hypoglycemic clamp in the absence of changes in counterregulatory hormones. Given formoterol's relatively rapid and long duration of action in the clinical setting, it could potentially be a useful therapeutic agent for prevention of hypoglycemia in patients with T1DM.
INTEGRATIVE HIGH-THROUGHPUT GENOMIC, EXPRESSION, AND EPIGENOMIC SEQUENCING OF INVASIVE VERSUS NONINVASIVE PITUITARY ADENOMAS: A CROSS-SECTIONAL STUDY
Zada G1, Pease M1, Mack W1, Weisenberger D1, Wang K1, Laird P1, Gruber S1 1USC, Los Angeles, CA, USA
OBJECTIVES/SPECIFIC AIMS: Pituitary Adenomas (PAs) are among the most commonly occurring intracranial tumors. PA invasion into surrounding anatomical structures is the most challenging clinical barrier to optimizing clinical outcomes, yet is not explained by somatic DNA mutations. Instead, PA invasion may be explained by differential gene expression due to epigenetic modification or copy number alterations. We aim to identify gene expression and/or methylation markers associated with tumor invasion. METHODS/STUDY POPULATION: 27 fresh-frozen surgically-resected PA specimens were graded with regard to tumor invasion based on the Knosp criteria (1993) in a blinded fashion by a collaborating neuroradiologist. PA specimens underwent DNA and RNA extraction followed by ongoing integrative genomic sequencing consisting of RNA sequencing, DNA methylation sequencing, DNA SNP array, and DNA exome sequencing. The distribution of Knosp scores in the 27 PAs were as follows: Grade 0 (n = 6), Grade 1 (n = 7), Grade 2 (n = 4), Grade 3 (n = 5), and Grade 4 (n = 5). Bioinformatics analysis will be used to compare noninvasive (Knosp score 0–1, n = 13) with invasive (Knosp score 2–4, n = 14) PAs. RESULTS/ANTICIPATED RESULTS: We hypothesize that invasive PA samples derived from patients who underwent pituitary surgery at Keck USC between 2009–2012, and subsequently assessed via integrative DNA methylation and RNA sequencing, will demonstrate hypermethylation, decreased gene expression, and gene silencing of tumor suppressor genes, as compared to noninvasive PA samples. DISCUSSION/SIGNIFICANCE OF IMPACT: This T1 phase cross-sectional pilot study will utilize next-generation DNA methylation and RNA sequencing platforms to identify specific genes and gene expression features correlated with tumor invasion, as a basis for future diagnostic and/or therapeutic intervention.
INTERLEUKIN-21 DRIVES INTESTINAL INFLAMMATION BY BRIDGING THE ADAPTIVE AND INNATE IMMUNE COMPARTMENTS
Poholek CH1, Harrington LE1 1University of Alabama at Birmingham, Birmingham, AL, USA
OBJECTIVES/SPECIFIC AIMS: Th17 CD4 T cells have been cast as pathogenic in the context of many autoimmune diseases, including Inflammatory Bowel Disease (IBD); however, the mechanism for this pathogenicity has yet to be elucidated. Based on the observation that IL-21 expression is increased in biopsies from patients with ulcerative colitis compared to healthy controls and Genome Wide Association Studies have shown an association between the locus containing il2/il21 and IBD, we posit that IL-21 produced by Th17 cells mediates inflammation in the intestine. METHODS/STUDY POPULATION: We use mouse models of both spontaneous and CD4 T cell-dependent colitis to interrogate the role of IL-21 signaling on disease pathogenesis. Analysis is completed using a combination of flow cytometry, ELISA, quantitative-PCR, and histological techniques. RESULTS/ANTICIPATED RESULTS: We show that a large number of IL-21-producing CD4 T cells are present in the intestines of mice with colitis and importantly, that IL-21 production is required for full disease progression. We find that although IL-21−/− CD4 T cells are unable to initiate disease, they produce elevated levels of IL-17A and IL-17F and are not selectively converted into Foxp3+ regulatory T cells, indicating that IL-21 is both crucial to the induction of colitis and surprisingly, is not acting via effects on the activation and differentiation of T cells. In direct support of this idea, IL-21R−/− CD4 T cells are capable of inducing disease unlike their IL-21−/− counterparts. In fact, our data demonstrate that IL-21 has an impact on the innate lymphoid cell compartment, which has previously been implicated in IBD pathogenesis. DISCUSSION/SIGNIFICANCE OF IMPACT: Taken together, our data show an important and previously unrecognized role for IL-21 that links CD4 T cells to innate lymphoid cells in the induction of chronic intestinal inflammation.
INVESTIGATION OF ANTI-BRONCHOCONSTRICTIVE PROPERTIES OF SOPHORA FLAVESENCE ALKALOIDS
Srivastava K1, Sampson HA1, Li X1 1The Mount Sinai School of Medicine, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: We wish to determine the efficacy and mechanism of action of Sophora flavesence alkaloid mediated inhibition of airway constriction. METHODS/STUDY POPULATION: We will use a murine model of asthma to evaluate effects of Sophora flavesence alkaloids on airway hyperreactivity using invasive spirometry of in vivo treated asthmatic mice and ex vivo experiments utilizing myography of murine tracheal rings. Ex vivo studies with human bronchial rings are also planned. RESULTS/ANTICIPATED RESULTS: In preliminary results we have found that Sophora flavesence alkaloids are potent suppressors of acetylcholine induced contraction of murine tracheal rings (P < 0.001 vs vehicle). We anticipate that Sophora flavesence alkaloid activity will be independent of beta-adrenergic agonist activity, sensitive to COX-inhibition and mediated by PGE2 as we have observed previously for the parent anti-asthma TCM formula ASHMI. We also expect that human bronchial tissue will be responsive to Sophora flavesence alkaloid activity. DISCUSSION/SIGNIFICANCE OF IMPACT: We believe that Sophora flavesence alkaloids have potential for development as asthma drugs for human use providing a non-beta adrenergic pathway for relief from bronchoconstriction.
INVESTIGATION OF RETT SYNDROME USING HUMAN IPSCS
Park I1, Tanaka Y1, Kim K1 1Yale University, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Rett syndrome (RTT) is one of the most prevalent female neurodevelopmental disorders that cause severe mental retardation. Mutations in methyl CpG binding protein 2 (MeCP2) that is present in Xq28 locus are mainly responsible for RTT. Following the normal development until 6 – 18 months, classic RTT patients show the symptoms, such as loss of language and motor milestone, purposeful hand movement, and normal head growth. Using reprogramming technology, we isolate induced pluripotent stem (iPS) cells from RTT patients' somatic cells. This study aims to investigate the genetic regulation of MeCP2 in pluripotent stem cells. METHODS/STUDY POPULATION: iPSC clones that we previously isolated express either wildtype MeCP2 (RTT-wt-iPSCs), mutated MeCP2 (RTT-mu-iPSCs) or both (RTT-bi-iPSCs) depending on the status of X chromosome. We applied massively parallel sequencing of RNA (RNA-seq) of RTT-iPSCs, normal iPSCs, and human embryonic stem cells (hESCs) and performed the comparative analysis. RESULTS/ANTICIPATED RESULTS: Consistent with the previous global gene expression analysis, the difference between hESCs and iPSCs was subtle, regardless of MeCP2 status. Surprisingly, our data showed that some sets of genes are differentially expressed between mutant RTT-iPSCs and wild type iPSCs, although the expression of MeCP2 is relatively lower in pluripotent stem cells compared with neurons. In addition to genes involved in neuronal development or function, MeCP2 mutant RTT-iPSCs showed the up-regulation of mitochondria-related genes. Furthermore, comparative analysis among five RTT patients revealed that distinct genes are affected by different mutations in MeCP2. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results show that MeCP2 regulates the physiologically fundamental genes from very early embryonic development, suggesting the importance in investigating the role of MeCP2 in diverse cell types.
ISOLATION OF IPRGC CONTRIBUTION TO THE HUMAN PUPILLARY LIGHT REFLEX
Hartwick A1, Yuhas P1 1Ohio State University, Columbus, OH, USA
OBJECTIVES/SPECIFIC AIMS: Intrinsically photosensitive retinal ganglion cells (ipRGCs), which express melanopsin photopigment, respond sluggishly to light and contribute to the sustained pupillary constriction that occurs after the offset of a bright stimulus. Due to the unique temporal properties of these ganglion cell photoreceptors, our aim was to utilize flickering light stimuli to isolate an ipRGC component to the human pupillary light reflex (PLR). METHODS/STUDY POPULATION: A flickering LED light stimulus was presented to the dilated left eye of 18 healthy subjects (age: 23 to 27). The consensual pupil response was recorded in the right eye under infrared illumination. The amplitude of the flicker in the pupil response (% of maximum pupil constriction) was determined using Fourier analysis. The effect of stimulus frequency (0.05 to 1.00 Hz), intensity (range: 1012 to 1015 phots/s/cm2), and prior light exposure on the pupil response to flickering red and blue light was determined. RESULTS/ANTICIPATED RESULTS: In response to a slowly flickering (0.1 Hz) stimulus, there was a significant reduction in magnitude of pupil flicker elicted by the blue versus red light as the intensity increased to 1014 phots/s/cm2. For flickering stimuli at 1014 phots/s/cm2, there was a significant difference in the pupil response elicited by the two stimuli (blue vs. red) when presented at 0.2, 0.1 and 0.05 Hz, but not at 0.5 or 1.0 Hz, flicker frequencies. Unexpectedly, the flicker in the pupil response to flashing (0.1 Hz) bright (1014 phots/s/cm2) red light significantly decreased when the red light was alternated with flashes of blue light as compared to the flickering stimuli of red light alone. DISCUSSION/SIGNIFICANCE OF IMPACT: The ipRGC input to the PLR can be best isolated with a bright, slowly flickering, stimulus that alternates between long and short wavelengths. This pupil testing strategy may have merit as a new clinical test of optic nerve function.
IVERMECTIN (IVM) DOSAGES THAT PRODUCE ANTI-ALCOHOL EFFECTS IN MICE TRANSLATE TO DOSES SAFE FOR USE IN HUMANS
Yardley MM1, Huynh N1, Louie SG1, Neely M1, Alkana RL1, Davies DL1 1University of Southern California, Los Angeles, CA, USA
OBJECTIVES/SPECIFIC AIMS: Alcohol use disorders (AUDs) represent a serious health issue. Our laboratory found that ivermectin (IVM), an FDA approved anthelmintic agent, can significantly reduce ethanol intake in mice following acute and chronic administration (Yardley, 2012 Neuropharmacology). Pharmacokinetic (PK) murine studies revealed an inverse relationship between the concentration of IVM in the brain in relation to the degree of ethanol intake. The current study begins to evaluate the PK and pharmacodynamic (PD) relationship of IVM in longer term alcohol studies and to determine if doses of IVM used in murine models would translate to doses approved for use in humans. METHODS/STUDY POPULATION: We developed a population PK model using single-dose IVM data from our murine model. We then tested the effect of IVM (3.0 mg/kg) on 10% v/v alcohol (10 E) intake using a 24-h two bottle choice model. IVM was administered (i.p.) daily for 10 consecutive days. On the 10th day, mice were euthanized at various time points and brain tissue and plasma samples were collected. Once we established the PK/PD model that best described the observed IVM concentrations and effects on ethanol consumption, we calculated the human dose range most likely to replicate the ideal exposure target in mice. RESULTS/ANTICIPATED RESULTS: Our initial findings show that the peak plasma concentration for the 2.5 mg/kg single dose is 50 ng/mL in mice. The human peak after a single dose of 30 mg is 85 ng/mL (Guzzo, 2002) corresponding to a murine dose of about 3.1 mg/kg. DISCUSSION/SIGNIFICANCE OF IMPACT: Initial results suggest that IVM dosages that produce anti-alcohol effects in mice correspond to doses already shown to be safe in humans. The findings lay the groundwork for advancing the testing of IVM as an anti-alcohol agent into humans.
LAPAROTOMY IN MICE INDUCES DIFFERENTIAL EXPRESSION OF INFLAMMATORY AND STRESS GENES
Ko FC1, Isoda F1, Mobbs C1 1Mount Sinai School of Medicine, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: Although surgical trauma and anesthesia alter immune and stress responses, their effects on post-surgical inflammatory and stress (oxidative stress, heat shock, growth arrest, DNA repair) genes remain poorly characterized. This pilot study sought to investigate the effect of laparotomy and isoflurane (a commonly used anesthetic) in mouse blood cell inflammatory and stress gene expression. METHODS/STUDY POPULATION: 12-week-old male C57BL/6J mice were subjected to 3 different interventions: 8 mice received 2% isoflurane via face mask; 8 mice received 2% isoflurane with laparotomy (1 cm vertical mid-abdominal incision and closure); and 8 mice received intraperitoneal saline injection (negative control). Mice were sacrificed 4 hours post-intervention and whole blood was collected for gene expression experiments. Using qPCR arrays, expression of 160 genes central to inflammatory and stress responses were measured. RESULTS/ANTICIPATED RESULTS: Mice subjected to isoflurane (15.6 ± 0.3 min) with laparotomy (8.5 ± 0.2 min), compared to mice subjected to isoflurane (15.4 ± 0.3 min) alone, showed >2 fold up-regulation (ΔΔCt, P < 0.007) of genes involved in inflammation (Osm, IL1rn, IL1b, Csf1), oxidative stress (Hmox1), heat shock (Hspa1b), growth arrest (Cdkn1a), and DNA repair (Ugt1a2). Moreover, expression of all 8 genes were significantly positively correlated (Pearson's correlation) with >50% of these correlations having ρ > 0.70 (P < 0.001). DISCUSSION/SIGNIFICANCE OF IMPACT: Surgical trauma from laparotomy differentially induces, and may co-regulate, post-surgical blood cell expression of specific inflammatory and stress genes. The roles of these genes in influencing surgical outcomes after laparotomy need to be further investigated.
MAPPING THE COGNITIVE CONNECTOME: TRANSLATING FUNCTIONAL NEUROIMAGING INTO PERSONALIZED MEDICINE
James GA1, Fausett J1, Gess J1, Kearney-Ramos T1, Reno A1, Peraza J1, Greenfield J1, Kilts C1 1University of Arkansas for Medical Sciences, Little Rock, AR, USA
OBJECTIVES/SPECIFIC AIMS: Our incomplete understanding of how the brain encodes cognitive variance remains the greatest barrier to the clinical translation of functional neuroimaging. Altered patterns of neural function have been reported for dozens of clinical disorders, but translating these findings into an individual patient's care requires a comprehensive mapping of individual variance in neurocognitive function for both healthy and clinical populations. To address this issue, we have developed the Cognitive Connectome-a comprehensive exploration of normative variability in cognition and brain function. METHODS/STUDY POPULATION: The Cognitive Connectome is built from a battery of well-validated neuropsychological tests and canonical functional MRI tasks spanning 8 cognitive modalities: motor, visuospatial awareness, attention, language, memory, affective processing, decision making, and executive function. Our pilot sample of 30 participants (mean ± sd age = 32 ± 10 years; 17 female; 11 African-American, 19 Caucasian) is allowing unprecedented insight into the nuanced interaction of brain and behavior. RESULTS/ANTICIPATED RESULTS: Our findings include the mapping of working memory's neural correlates and its supervisory influence upon other cognitive modalities and their brain networks; a demonstration of how personality factors influence the processing of novel emotional stimuli; and relating individual differences in cognition to individual differences in brain connectivity patterns. DISCUSSION/SIGNIFICANCE OF IMPACT: Applications into personalized medicine will emerge as this sample grows – i.e. predicting patient's treatment response from baseline brain scans or neurosurgical pre-planning to avoid cognitive impairment. Acknowledgments: This research supported by the CTSA (8 UL1 TR000039–04) and KL2 Scholars Program (8 KL2 TR000063-04).
MEASUREMENTS OF CEREBRAL BLOOD FLOW DURING HYPOGLYCEMIA IN SUBJECTS WITH TYPE 1 DIABETES AND HYPOGLYCEMIA UNAWARENESS
Mangia S1, Tesfaye N1, De Martino F1, Kumar AF1, Kollasch P1, Moheet AA1, Eberly LE1, Seaquist ER1 1University of Minnesota, Minneapolis, MN, USA
OBJECTIVES/SPECIFIC AIMS: Hypoglycemia (HG) is a serious complication of diabetes regimens based on insulin or insulin secretagogues. The thalamus has been found in previous studies to be activated during the early phase of moderate HG. Here, we tested the hypothesis that this region is less activated during HG in HG-unaware subjects with type 1 diabetes (T1DM) relative to controls. Cerebral activations were detected in the midbrain regions based on changes in regional cerebral blood flow (CBF). METHODS/STUDY POPULATION: Twelve controls (5 F/7 M, age 40 ± 14 years, body mass index 24.2 ± 2.7 kg/m2) and eleven patients (7 F/4 M, age 39 ± 13 years, body mass index 26.5 ± 4.4 kg/m2) with well-controlled T1DM (A1c 6.8 ± 0.4%) underwent a two-step hyperinsulinemic clamp while laying inside a 3 Tesla scanner. Blood glucose was held at 95 mg/dL and then allowed to decrease to 50 mg/dL. CBF weighted images were acquired using arterial spin labeling during euglycemia and HG. RESULTS/ANTICIPATED RESULTS: The HG-induced CBF changes in the thalamus of healthy controls were significantly greater as compared with those of HG-unaware T1DM subjects. A significant correlation was also observed between thalamic activations and the within-person epinephrine response during HG. DISCUSSION/SIGNIFICANCE OF IMPACT: Our findings suggest that thalamic activation has a critical role in the detection of HG and the coordination of the sympathetic CR response necessary to restore normoglycemia. We are currently undertaking further studies to ascertain the distinct roles of diabetes and HG unawareness in the abnormal brain responses. This research is essential for designing drugs and therapies that target the brain mechanisms responsible for HG unawareness, and therefore has large impact in reducing the complications of management of diabetes.
MICE WITH SICKLE CELL DISEASE HAVE DECREASED ASCORBIC ACID TISSUE LEVELS
Chheath M2,1, Almeida L2,1, Quezado Z2,1 1Children's National Medical Center, Washignton, DC, USA; 2The Sheikh Zayed Institute for Pediatric Surgical Innovation, Washington, DC, USA
OBJECTIVES/SPECIFIC AIMS: Some of the complications of sickle cell disease (SCD) are believed to be associated with and/or result from increased oxidative stress. Ascorbic acid is known to be an important antioxidant due to its redox potential as a reducing agent and a cofactor in several enzyme reactions and collagen production. We hypothesized that mice with SCD have altered tissue level of ascorbic acid. Using a mouse model known to recapitulate several complications of human SCD, we examined the effect of SCD on tissue concentration of ascorbic acid and evaluated the effectiveness of ascorbic acid diet supplementation. METHODS/STUDY POPULATION: Using an ELISA assay we measured tissue concentration of ascorbic acid in SCD mice RESULTS/ANTICIPATED RESULTS: We found that homozygous (a model for SC anemia) had lower concentrations of ascorbic acid in liver and kidney (p < 0.001) compared to controls. Heterozygous male mice (a model of SC trait) had lower concentrations of ascorbic acid in liver compared to controls (p < 0.001). Interestingly, these deficiencies observed in homozygous and heterozygous SCD male mice were not found in female mice. Further, when SCD mice received supplemental ascorbic acid in water (0.04%, the human equivalent of 1 g/day) for one week, hepatic and renal tissue levels of ascorbic acid in male SCD mice were similar to control mice. DISCUSSION/SIGNIFICANCE OF IMPACT: Our findings suggest that SCD is associated with decreased concentrations of tissue ascorbic acid. One can postulate that ascorbic acid deficiency could contribute to oxidative stress in mice with SCD and in turn to organ dysfunction. Further, our findings also suggest that the degree of ascorbic acid deficiency depends on sex and genotype. Extrapolating our findings clinically, ascorbic acid deficiency might contribute to morbidity in patients with SCD.
MODIFIED NUCLEIC ACIDS: A THERAPEUTIC ALTERNATIVE IN CHRONIC S. AUREUS INFECTIONS
Atwood DN1, Beenken KE1, Griffin LM1, Mrak LN1, Zielinska AK1, Matthews KA1, Smeltzer MS1 1University of Arkansas for Medical Sciences, Little Rock, AR, USA
OBJECTIVES/SPECIFIC AIMS: Staphylococcus aureus causes a diverse array of infections. Treatment is compromised by the emergence of antibiotic-resistant strains, including MRSA, even among community-acquired isolates. However, many types of S. aureus infections are recalcitrant to antimicrobial therapy even in the absence of conventional antibiotic resistance. A primary reason for this is formation of a biofilm, which confers intrinsic resistance to both host defenses and all currently available antibiotics. We have confirmed that mutation of global regulator sarA limits biofilm formation to a degree that can potentially be exploited to overcome this intrinsic resistance. As such, we are investigating the use of modified nucleic acids, including morpholinos and peptide nucleic acids, to inhibit the production and/or function of sarA. METHODS/STUDY POPULATION: With respect to production, we are employing an anti-sense approach targeting the ribosomal binding site of the sarA mRNA transcript, thereby limiting the production of SarA itself. Additionally, we have demonstrated that a primary function of SarA is to repress the production of extracellular proteases, thereby allowing the accumulation of critical virulence factors including those required for biofilm formation. Based on this, we will also employ an anti-target approach aimed at limiting the interaction between the SarA DNA-binding protein and its cis targets associated with the genes encoding critical extracellular proteases. RESULTS/ANTICIPATED RESULTS: We believe inhibition of SarA will lead to an increased production of proteases, resulting in decreased biofilm formation. DISCUSSION/SIGNIFICANCE OF IMPACT: It is hoped that this combined approach will allow us to take therapeutic advantage of SarA to overcome the intrinsic resistance that defines biofilm-associated S. aureus infections.
NEAR-INFRARED FLUORESCENCE IMAGING OF LYMPH NODE METASTASES BY TARGETING GASTRIN RELEASING PEPTIDE RECEPTOR
Cai Q1,4, Yu P2, Hoffman TJ3,4, Ma L1,4 1Departments of Radiology, University of Missouri, Columbia, MO, USA; 2Physics and Astronomy, University of Missouri, Columbia, MO, USA; 3Internal Medicine, Chemistry, University of Missouri, Columbia, MO, USA; 4Harry S. Truman VA Hospital, Columbia, MO, USA
OBJECTIVES/SPECIFIC AIMS: The current accuracy for lymph node (LN) staging in cancer patients is low. We herein report a new study on the design and use of near-infrared fluorescence (NIRF)-labeled bombesin (BBN) probes as tumor-targeting molecular imaging agents for specific detection of LN and systematic metastases in prostate cancer. METHODS/STUDY POPULATION: Alexa Fluor 680 conjugated BBN[7–14]NH2 (AF680-BBN) peptides, which show efficient NIRF emission under an excitation, were synthesized. AF680-BBN was administered intravenously in a PC-3/Luc+ orthotopic mouse model of human prostate cancer. LN and peritoneal metastases were detected by NIRF imaging and confirmed by histopathology. RESULTS/ANTICIPATED RESULTS: Tumor-to-background ratio (TBR) in the NIRF imaging reached the highest value at two-hour (4.12 ± 1.77) post-injection of AF680-BBN. Blocking experiments, using unlabeled BBN as the inhibiting agent, significantly reduced the TBR (1.64 ± 0.21, p = 0.02). The imaging sensitivity and specificity were assessed by co-registration of NIRF imaging and luciferase bioluminescence imaging of the mice. AF680-BBN was shown to have a sensitivity of 89.4%, specificity of 92.9%, and accuracy of 90.2% for the detection of metastases in mice. DISCUSSION/SIGNIFICANCE OF IMPACT: The studies suggest the potential use and development of NIRF-labeled BBN probes as cancer-specific agents. In clinical practice, a laparoscope incorporated with a NIRF camera could provide both NIRF and white light imaging of the surgical field during the intra-operative procedure. Our developments could help improve the current detection and lymph node staging accuracy, or provide intra-operative imaging guidance in prostate cancer.
NEURAL MECHANISMS OF SWITCHING ATTENTIONAL STATE IN OBSESSIVE-COMPULSIVE DISORDER
Stern ER1, Muratore AF1, Taylor SF2, Abelson JL2, Hof PR1, Goodman WK1 1Mount Sinai School of Medicine, New York, NY, USA; 2University of Michigan, Ann Arbor, MI, USA
OBJECTIVES/SPECIFIC AIMS: Obsessive-compulsive disorder (OCD) is associated with excessive absorption in distressing thoughts, images, or feelings, which may be due to an inability to flexibly switch attention away from an internally focused state toward external information in the environment. Functional magnetic resonance imaging (fMRI) studies have linked internally focused cognition such as event imagination to activation of the “default mode network” (DMN). The current study investigated activation of the DMN when patients and controls were required to switch attention from internal to external information. METHODS/STUDY POPULATION: Preliminary fMRI data was obtained from 12 healthy individuals and 4 OCD patients in a novel switching task where subjects imagined personal event scenarios before switching to perform an externally focused target detection task (TD). Whole-brain analyses compared activation patterns in OCD patients to controls when subjects must disengage from event imagination to perform TD. RESULTS/ANTICIPATED RESULTS: As expected, internally focused event imagination activated DMN regions in the healthy control group and in both OCD patients. When required to switch attention from an internal to an external focus, the healthy control group ceased to exhibit DMN activity, whereas OCD patients continued to activate frontal and temporal regions of DMN. DISCUSSION/SIGNIFICANCE OF IMPACT: Although very preliminary, these findings suggest that OCD is associated with a failure to disengage brain systems involved in internally focused mental processes when attention must be switched toward external information.
NEURAL REORGANIZATION OF LANGUAGE FOLLOWING A NOVEL INTERVENTION FOR PEOPLE WITH APHASIA
Dietz A1, Vannest J2, Holland S2, Szaflarski J3,1 1University of Cincinnati, Cincinnati, OH, USA; 2Cincinnati Children's Medical Center and Hospital, Cincinnati, OH, USA; 3University of Alabama-Birmingham, Birmingham, AL, USA
OBJECTIVES/SPECIFIC AIMS: AIM 1: To generate pilot data regarding the effect of a high-technology augmentative and alternative (HITAAC) treatment on the restoration of language and communication skills in people with aphasia (PWA). AIM 2: To generate pilot functional magnetic resonance imaging (fMRI) data demonstrating intervention-related reorganization in the neural pathways underlying improved language and communication skills in response to a HITAAC Treatment in PWA. METHODS/STUDY POPULATION: The participants will receive 4 weeks of HITAAC treatment. Discourse assessment and fMRI testing will occur pre-treatment, post-treatment, and 3 months post-treatment to document baseline and treatment-related changes. RESULTS/ANTICIPATED RESULTS: AIM 1: The HITAAC Tx/Personalized discourse instruction will lead to improved language and communication abilities in PWA, assessed by improvement on generally accepted measures of discourse. AIM 2: HITAAC Tx/Personalized discourse instruction will result in changes in cortical mechanisms that are associated with improved performance on generally accepted measures of discourse. DISCUSSION/SIGNIFICANCE OF IMPACT: This study will provide preliminary behavioral and fMRI data that AAC interventions facilitate language recovery and lay the groundwork for the development for a more efficient and effective approach to aphasia rehabilitation.
NEUROENDOCRINE AND PSYCHOLOGICAL STRESS SYMPTOMS IN CHILDREN AND PARENTS AFTER ADMISSION TO THE PEDIATRIC INTENSIVE CARE UNIT
Nelson LP1, Young CM1, Gold JI1 1Keck School of Medicine, University of Southern California, Los Angeles, CA, USA
OBJECTIVES/SPECIFIC AIMS: Improved survival outcomes of critical illness require a focus on longterm outcomes following admission to a pediatric intensive care unit (PICU). Given the effects of stress on functional outcomes, PICU-related stress is a significant concern. We hypothesize children and their parents with higher neuroendocrine markers of stress during PICU admission will have increased acute stress disorder (ASD) symptoms at follow up. METHODS/STUDY POPULATION: This is a prospective observational study in an academic PICU. Subjects are English- and Spanish- speaking children (ages 8–17 years) and parents admitted with a predicted stay of > 48 hours. Exclusion criteria include developmental delay, psychiatric disorder, intentional injury, traumatic brain injury, or inability to complete measures. Assessments are done at baseline (T1) and 1 month after admission (T2), and include cortisol levels and the Acute Symptoms Checklist for Children (ASC-Kids)/Stanford Acute Stress Reaction Questionnaire (SASRQ). RESULTS/ANTICIPATED RESULTS: Children (N = 14; M Age = 14 years; 10 male) and their parents (N = 14; M Age = 40 years; Spanish-speaking = 7) were recruited. 36% of children were not on steroids; their T1 cortisol and T2 ASD symptoms have a positive correlation (r = .84, p = .079). Parents' T1 cortisol and T2 ASD symptoms have a positive relationship, but is not significant (r = .42, p = .40). DISCUSSION/SIGNIFICANCE OF IMPACT: While the association between children' neuroendocrine markers of stress during PICU admission and later ASD symptoms is approaching significance, the parents' is not significant. This study is encouraging in its association between stress symptoms during and following PICU admission, but is limited by a small sample size. This investigative team continues to recruit subjects.
NEUROENDOCRINE IMMUNE REGULATION IN MELANOMA: IMPLICATIONS FOR NOVEL THERAPEUTIC STRATEGIES
Dronca R1, Leontovich A1, Nevala W1, Markovic S1 1Mayo Clinic, Rochester, MN, USA
OBJECTIVES/SPECIFIC AIMS: The hypothesis of our study is that immune system dynamics in metastatic melanoma (MM) can be therapeutically utilized to increase the efficacy of conventional chemotherapeutic drugs, and that host/tumor immune interactions are regulated at systemic level by conventional (tumor) as well as non-conventional (neuroendocrine) mechanisms. METHODS/STUDY POPULATION: Patients with MM scheduled to undergo treatment with temozolomide (TMZ) undergo immunological monitoring consisting of 10 daily blood collections prior to initiation of therapy. At the end of the collection period, samples are batched analyzed for dynamic changes in immune cells and plasma cytokines and an optimal day for initiation of TMZ based on individualized summary immune dynamics is prospectively calculated. RESULTS/ANTICIPATED RESULTS: Systemic immunity in MM follows mathematically predictable temporal kinetics, which can be utilized to selectively enhance cytotoxic T-cell responses and/or deplete immune-suppressor cells in a cycle-specific (time-dependent) manner. The interim analysis of stage I patients suggests clinical improvement with a 4-month progression-free survival (PFS) rate of 60% compared to 20% conventional therapy (TMZ random delivery) historical rate. These recurring oscillations of systemic immunity between states of up and down regulation reflect the host immune response to cancer, as well as a manifestation of neuroendocrine regulation of immunity. DISCUSSION/SIGNIFICANCE OF IMPACT: The translational value of this study is identification of novel approaches to increase the efficacy of therapy in MM through individualized treatment administration. Moreover, understanding sex-based differences of immunity in cancer may have applicability to other tumor types and enable identification of patients with non-classical hormone sensitive tumors who may benefit from addition of hormone-targeted treatment.
NITRIC OXIDE RELEASING POLYMERIC COATINGS FOR PREVENTION OF THROMBOSIS AND INFECTION IN SHORT-TERM AND LONG-TERM ANIMAL MODELS
Handa H1, Brisbois E1, Major T1, Annich G1, Meyerhoff M1, Bartlett R1 1University of Michigan, Ann Arbor, MI, USA
OBJECTIVES/SPECIFIC AIMS: 1) To fabricate and optimize nitric oxide (NO) release coatings that can sustain the release of NO for up to 2 weeks. 2) To evaluate the effect of NO release (NORel) coatings in various animal models for prevention of thrombosis and infection. METHODS/STUDY POPULATION: Various polymer formulations were studied to obtain an NO release for up to 2 weeks. The NORel coatings were evaluated for prevention of thrombus and infection in a short-term (4 h) and a long-term (>10 d) rabbit model. RESULTS/ANTICIPATED RESULTS: The NORel coatings containing the pH control additive continuously release 7–18 × 10−10 mol/cm2-min NO for 14 d at 37°C in PBS buffer. Four out of 7 control circuits were clotted within 3 h. Platelet counts after 4 h on the NORel ECC were preserved at 94% of the baseline count compared to 47% preservation on control circuits. NORel catheters are expected to show a significant decrease in thrombus and biofilm formation as compared to the corresponding controls in a long-term animal model. DISCUSSION/SIGNIFICANCE OF IMPACT: This study demonstrates, by controlling the pH inside that NO release polymeric coatings containing lipophilic dizeniumdiolate species, sustains the NO release for > 2 weeks. A significant reduction in the clot area was seen as compared to the control 4 h ECC circuits. These NORel coatings are now under investigation in a 2–3 week rabbit model and could provide a breakthrough in long-term preservation of circulating platelets.
NOVEL POLY-LACTIC ACID NANOFIBER-FILLED SLURRY FOR IMPROVED ELASTIC CARTILAGE ENGINEERING
Halevi AE1, Caballero M1, Wright MC2, Pharaon M1, van Aalst JA1 1University of North Carolina, Chapel Hill, NC, USA; 2Xanofi, Inc, Raleigh, NC, USA
OBJECTIVES/SPECIFIC AIMS: Synthetic nanofiber scaffolds promote cell attachment, proliferation, and differentiation, offering a dynamic platform for elastic cartilage engineering. A current limitation of electrospun materials is the lack of true 3-dimensionality. A novel glycerin-based method for creating nanofiber-filled slurries (Xanofi) composed of disassociated nanofibers creates a more true 3-dimensional (3-D) scaffold for improved cell-scaffold interaction. In this study, human umbilical cord mesenchymal stem cells (UCMSCs) are utilized to demonstrate cell attachment, proliferation, and chondrogenesis in a poly-lactic acid nanofiber-filled slurry (PLANS). METHODS/STUDY POPULATION: hUCMSCs were isolated from Wharton's Jelly by explant technique, grown to 80% confluence, and seeded at passage 3 (1e4 cells/scaffold) on 3 material densities (0.05, 0.1, 0.5 g/well) of PLANS (1.28% solids) and grown for 7 days in standard medium. Cells were stained with calcein/ethydium bromide, DAPI, and imaged by fluorescent microscopy; DNA was collected on days 1, 3, 5, and 7. Cells were chondroinduced for 10 days with TGF-beta-1, TGF-beta-3, and IGF. Samples were fixed and stained for glucosaminoglycan (GAG) content; GAG was quantified using DMMB assay; collagen I, II, X, and elastin mRNA were determined by qRT-PCR at days 7 and 10. RESULTS/ANTICIPATED RESULTS: Cells were viable throughout the study. DNA quantification demonstrated MSC proliferation for the entire 7 days. The presence of GAG, increased collagen II and elastin mRNA, with no increase in collagen X, was consistent with elastic cartilage formation. DISCUSSION/SIGNIFICANCE OF IMPACT: PLANS is a novel material that facilitates cell viability, proliferation, and chondrogenesis, and may provide a superior platform for generating engineered elastic cartilage.
ORAL CONTRACEPTIVES USE AND THE RISK OF CARDIAC EVENTS IN LONG QT SYNDROME PATIENTS
Abu-Zeitone A1, Peterson DR1,2, McNitt S1, Polonsky S1, Moss AJ1 1University of Rochester Medical Center, Rochester, NY, USA; 2Department of Biostatistics & Computational Biology, Rochester, NY, USA
OBJECTIVES/SPECIFIC AIMS: To study the effect of using oral contraceptives on the risk of cardiac events (syncope/aborted cardiac arrest) in females with Long QT Syndrome (LQTS) METHODS/STUDY POPULATION: A subset of female patients from the LQTS Registry were selected to receive questionnaires about their history of oral contraceptive use (n = 340). Time-dependent Cox regression analysis will be used to estimate the hazard ratio of cardiac events (CE) when patients are ON versus OFF oral contraceptives during non-pregnancy and non-postpartum periods. For this analysis, we defined time origin as the onset of menarche and we will be adjusting for these variables: age at onset of menarche, baseline QT interval, history of prior CE and therapy used. When pregnancy occurs, patients will be censored for 2 years to account for changes in the risk of CE during pregnancy, postpartum and breastfeeding periods. RESULTS/ANTICIPATED RESULTS: We anticipate that there will be fewer cardiac events during the time patients are receiving versus not receiving oral contraceptives. DISCUSSION/SIGNIFICANCE OF IMPACT: This study will be the first to examine the effect of oral contraceptives on the risk of CE in LQTS patients. Previous LQTS animal studies found that high progesterone concentrations have favorable effects compared to estrogen. Similarly LQTS clinical studies found significant reduction in CE during pregnancy, when high progesterone/estrogen ratio is achieved physiologically. Oral contraceptive pills represent a similar clinical model with high progesterone/estrogen ratio. Therefore, if this study shows a reduced risk of CE in LQTS, it will: 1) support recent LQTS animal studies 2) translate into new treatment strategies (progesterone) and 3) provide new information for future research.
OREXIN RECEPTOR ANTAGONISTS ATTENUATE HOT FLASH-LIKE RESPONSES IN A OVARIECTOMY MODEL OF MENOPAUSE-ASSOCIATED SYMPTOMS
Johnson PL1, Federici LM1, Skaar T1, Carpenter J1, Renger JJ2, Gotter AL2, Winrow CJ2, Samuels BC1, Shekhar A1 1Indiana University School of Medicine, Indianapolis, IN, USA; 2Merck Research Laboratories, West Point, PA, USA
OBJECTIVES/SPECIFIC AIMS: Menopause and ovariectomy (OVEX) are conditions where estrogen loss leads to hot flashes. Estrogen replacement is an effective treatment, but with significant cardiovascular and cancer risks. Therefore there is a need for non-hormonal therapies, but the limited scientific understanding and lack of animal models of menopause symptoms has hindered progress. We developed an OVEX rat model of hot flash vulnerability to a GABA disenhancer (FG-7142), hypercapnic gas, and yohimbine. Convergent data shows that these inducers increase activity in only 3 brain regions, of which only the dorsomedial/perifornical hypothalamus (DMH/PeF) contains estrogen receptors, polysynaptically innervates tail, and is thermoregulatory. Since orexin synthesizing neurons are almost exclusive to the DMH/PeF and menopausal women have 300% higher central ORX levels which are rescued with estrogen replacement, we predicted that a hyperactive ORX system contributes to menopausal symptoms. METHODS/STUDY POPULATION: In order to test this hypothesis, we: 1) OVEXed female rats to model menopausal symptoms; 2) induced hot flash-like tail temp increases (∼4–7C) using a GABA disenhancer; then 3) screened centrally active single ORX1 (SORA) and ORX1/2 (DORA) receptor antagonists. RESULTS/ANTICIPATED RESULTS: Systemically pretreating OVEX rats with a SORA or DORA attenuated “hot flash”-associated responses. DISCUSSION/SIGNIFICANCE OF IMPACT: These results are the 1st to propose and support the hypotheses that: 1) loss of estrogen inhibition of the ORX system leads to menopause-related symptoms, and 2) that ORX1 or ORX1/2 receptor antagonists constitute a novel method of treatment.
PANNEXIN 1 AND ATP SIGNALING IN BLADDER FUNCTION AND DYSFUNCTION
Urban-Maldonado M1, Negoro H1, Tar MT1, Melman A1, Thi MM1, Suadicani SO1 1Albert Einstein College of Medicine, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: Urothelial cells respond to bladder distension with ATP release, and ATP signaling to the CNS and within the bladder relay the degree of bladder fullness and modulate detrusor activity, which are essential for proper micturition. In various cell types, Pannexin 1 (Panx1) channels provide a pathway for mechanically-induced ATP efflux and for ATP-induced ATP release via interaction with P2×7 receptors (P2×7Rs). We investigated whether Panx1 channels participate in bladder ATP signaling and whether Panx1 expression and function are altered in diabetic bladders. METHODS/STUDY POPULATION: Bladders from 2 month streptozotocin (STZ) diabetic and age-matched control rats were used. Panx1 and P2×7R expression and function were analyzed by qPCR, Western, immunostaining and ATP release. RESULTS/ANTICIPATED RESULTS: Panx1 and P2×7R are expressed in bladder urothelium, lamina propria (LP) and detrusor muscle. Bladder mucosa stimulation with BzATP (300 μM; P2×7R agonist) induced release of significant amounts of ATP (439.6 ± 57.8 vs 1.3 ± 0.1 nM; p < 0.001, N = 3) that was blunted (62.4 ± 12.3 vs 1.5 ± 0.1 nM, N = 3) by mefloquine (MFQ; 100 nM, Panx1 blocker). Intravesical ATP release induced by bladder distension was also significantly reduced by MFQ. In diabetic rats, Panx1 mRNA expression in the bladder urothelium, LP and detrusor was significantly higher than in control tissues. Consistent with Panx1 upregulation, intravesical ATP release induced by bladder distension was higher in bladders from diabetic rats. DISCUSSION/SIGNIFICANCE OF IMPACT: Panx1 channels provide a pathway for ATP release from bladder mucosa and Panx1 expression is upregulated in diabetic bladders. These findings indicate that Panx1 channels may play a key role in mechanisms involved in transduction of bladder distension and modulation of bladder function in physiological and pathological conditions. (Support: NIH DK081435).
PARACRINE EFFECTS OF ADIPOSE STROMAL CELL CONDITIONED MEDIA ON RETINA: A CELL-FREE THERAPY FOR RETINAL DISEASES
Rajashekhar G1, Abburi C1, Maturi R1, Harris A1, Kern T2, March K1 1Indiana Univ Sch of Medicine, Indianapolis, IN, USA; 2Case Western Reserve University, Cleveland, OH, USA
OBJECTIVES/SPECIFIC AIMS: Previously we have shown that adipose stromal cells (ASC) therapeutically rescue against vascular inflammation and apoptosis in diabetic retinopathy. In this study, we tested if adipose stromal cell conditioned media (ASC-CM) can protect retinal damage in-vivo and may serve as an alternate cell free therapeutic modality for retinal diseases. METHODS/STUDY POPULATION: Acute ischemia-reperfusion (I/R) injury and chronic streptozotocin-induced diabetes of 2-mos duration were tested. Intravitreal injection was performed with ASC-CM into the right eye (2 μL) and an equal volume of basal media into the left eye. Day-7 post injection, retinal function assessed by Electroretinogram (ERG), retinal histological changes were observed by light microscopy and vascular leakage by FITC-albumin extravasation method. RESULTS/ANTICIPATED RESULTS: I/R resulted in a significant reduction in peak “b” wave ERG amplitude compared to un-injured control rats, and this reduction was significantly improved by ASC-CM at day-7 post injection (p < 0.02; n = 6). Subsequently, retinal histological analyses at day-7 revealed an increase in retinal thickness; decrease in gliosis and decrease in apoptosis (for all p < 0.05, n = 5) in retina from injured eyes received ASC-CM compared to injured eyes received BME. In addition, in a chronic diabetic model, increased vascular leakage in diabetic animals were significantly reduced in diabetic animals that received ASC-CM (n = 5; p < 0.02) within 7 days. DISCUSSION/SIGNIFICANCE OF IMPACT: Taken together with both acute and chronic retinopathy models, our findings suggest that ASC-CM may serve as a cell free therapy against retinal pathology with its ability to down modulate gliosis, vascular permeability and neural cell death. Future studies will assess the possible beneficial proteins from ASC-CM.
PEPTIDE-MEDIATED INHIBITION OF MK2 AMELIORATES BLEOMYCIN-INDUCED LUNG FIBROSIS
Vittal R1, Fisher AJ1, Gu H1, Mickler EA1, Panitch A2,3, Lander C3, Cummings OW1, Sandusky GE1, Wilkes DS1 1Indiana University School of Medicine, Indianapolis, IN, USA; 2Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA; 3Moerae Matrix Inc., Morristown, NJ, USA
OBJECTIVES/SPECIFIC AIMS: To determine the effects of inhibiting MK2 (mitogen-activated protein kinase activated protein kinase 2;MAPKAPK2), a serine/threonine kinase downstream of p38MAPK, in alleviating lung fibrosis. METHODS/STUDY POPULATION: MK2 expression was determined in lung biopsies from patients with idiopathic pulmonary fibrosis (IPF). C57BL/6 mice received intra-tracheal bleomycin instillation (bleo; 0.025 U; day 0), followed by phosphate buffered saline (PBS) or MK2 inhibitor (MK2i; MMI-0100; 37.5 μg/kg), administered either via local (nebulized) or systemic (intra-peritoneal) routes. MK2i or PBS was dosed daily for 14 days post-bleo injury, beginning day 7 or day 14. Normal primary human fetal lung fibroblasts were treated ± MK2i ± TGF-beta and incubated for 72 h. RESULTS/ANTICIPATED RESULTS: Compared to pathologically normal lungs, the IPF lungs have higher MK2 activation localized to fibroblasts and epithelial cells. In the bleo model, we observed robust MK2 activation at day 7 (pre-fibrotic stage) and day 14 (post-fibrotic stage). Regardless of mode of administration or stage of intervention, MK2i abrogated collagen deposition, myofibroblast differentiation and MK2 activation. MK2i also decreased circulating TNF-α and IL-6 levels and modulated local mRNA expression of pro-fibrotic cytokine il-1β, matrix genes (col1a2, col3a1, lox) and TGF-beta family members (smad3/6/7, serpine1). In vitro, MK2i dose-dependently attenuated myofibroblast differentiation, collagen type I and fibronectin expression and activation of MK2 and FAK. DISCUSSION/SIGNIFICANCE OF IMPACT: Peptide mediated inhibition of MK2 impacts both inflammatory and fibrotic responses and thus may be a promising therapeutic target for IPF.
PERSONALITY AND EXPECTANCY MEASURES PREDICT RATES OF INTRAVENOUS ALCOHOL SELF-ADMINISTRATION IN SOCIAL DRINKERS
Zametkin M1, Stangl BL1, Vatsalya V1, Ramchandani VA1 1NIAAA, Bethesda, MD, USA
OBJECTIVES/SPECIFIC AIMS: Computer Assisted Self-Infusion of Ethanol (CASE) is a method of intravenous (IV) alcohol administration that provides individuals with access to ad lib self-infusions of alcohol in a laboratory setting. The objective of this study was to examine the effects of impulsivity and expectancy on IV alcohol self-administration behavior in social drinkers. METHODS/STUDY POPULATION: Social drinkers (N = 87) underwent a self-administration session that consisted of a 25 minute priming phase where subjects were prompted to push a button to receive individually standardized alcohol infusions, followed by a 2 hour “open bar” phase with ad-lib access to the same infusions. Participants were grouped based on the number of infusions they administered during the first 30 minutes of the session into 3 categories: non-, low, and high responders (0, 1–5, or 6–12 infusions, respectively). Impulsivity was measured using the Delayed Discounting task (DD). Expectancy was measured using the Alcohol Effects Questionnaire (AEFQ). RESULTS/ANTICIPATED RESULTS: Results indicated significant differences between non-, low and high responders for personality and expectancy measures. High responders had lower expectancy scores for power and aggression as well as cognitive and physical impairment (all p values < 0.05). High responders also had greater measures of impulsivity compared to non- and low responders (all p values < 0.05). DISCUSSION/SIGNIFICANCE OF IMPACT: Results indicate that higher rates of IV alcohol self-administration were associated with higher impulsivity, and lower expectancy of cognitive and physical impairment and aggression following alcohol. These findings suggest that the rate of IV alcohol self-administration is sensitive to the expected behavioral effects of alcohol in social drinkers.
PHARMACOLOGICAL CHARACTERISTICS OF BASS RIVER STATE FOREST PLANTS AND ITS APPLICATION IN IMMUNOLOGICAL PROPERTIES: OCCURRENCE FROM THE NEW JERSEY
Mollik M1,2 1Prescience Trust Funds, Phoenixville, PA, USA; 2Peoples Integrated Alliance, Dhaka, Bangladesh
OBJECTIVES/SPECIFIC AIMS: United States is one of the seventeen mega-diverse countries of the world having rich vegetation with a wide varies; scientific studies of plants have been initiated because of their contribution to healthcare. METHODS/STUDY POPULATION: Extensive chromatographic separation and purification of extracts from the plants of Bass River State Forest in New Jersey have yielded a wide variety of structurally unique and biologically active secondary metabolites. They have been identified using various states and regional floras. RESULTS/ANTICIPATED RESULTS: These plants included Acalypha australis L., Alisma subcordatum Raf., Dioscorea oppositifolia L., Mentha aquatica L., Onoclea sensibilis L., Rumex britannica L., Sagittaria cuneata E.Sheld., and Thaspium barbinode (Michx.) Nutt.. Over the years it has been possible to isolate secondary metabolites like terpenes, steroids, flavonoids, megastigmanes, benzohydrofurans, phenylethanoids and their glycosides: saponins, free amino acids plus small molecular peptides, and soluble dietary fibres from the plant extracts. The structure elucidations of the isolated constituents were characterized by ultraviolet, nuclear magnetic resonance, and multiple sclerosis experiments. The studies showed that plant extracts are complex in nature which contains diversified molecules. Chemical constituents present in the active extracts were also found to be different in chemical nature. Biological testing showed wide variation in activities. In addition bio-assay studies have shown potential medicinal values of the plant extracts. DISCUSSION/SIGNIFICANCE OF IMPACT: The studies have been a strong evidence for the United States as a promising source of natural products. It is thought desirable to assess these claims on scientific basis and to authenticate thereby their efficacy and toxicity.
PLASMA METABOLOMIC ANALYSIS OF HIV-INFECTED WOMEN ASSOCIATES INCIDENT DIABETES MELLITUS WITH ESSENTIAL AMINO ACID METABOLISM
Aleman JO1, Hoover D1, Shi Q2, Kim P1, Kang J1, Anastos K3, Tien P4, Sharma A5, Nowicki M6, Cohen M7, Golub E8, Young M9, Glesby MJ1, Rhee KY1 1Weill Cornell Medical College, New York, NY, USA; 2New York Medical College, Valhalla, NY, USA; 3Montefiore Medical Center, Bronx, NY, USA; 4UCSF & San Francisco VA Medical Center, San Francisco, CA, USA; 5SUNY Downstate Medical Center, Brooklyn, NY, USA; 6University of Southern California, Los Angeles, CA, USA; 7Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA; 8Cook County Health & Hospitals System and Rush University, Chicago, IL, USA; 9Georgetown University Medical Center, Washington, DC, USA
OBJECTIVES/SPECIFIC AIMS: Several cohort studies, including the Women's Interagency HIV Study (WIHS), reported increased incidence of diabetes mellitus (DM) in HIV-infected subjects receiving combination antiretroviral therapy relative to HIV-uninfected controls. METHODS/STUDY POPULATION: This study sought to find a plasma signature of incident DM in predominantly HIV-infected WIHS women by a targeted metabolomic framework. Using Liquid Chromatography-Mass Spectrometry technology we analyzed a training set of fasting plasma samples from 17 women with incident DM (cases; samples drawn 1–2 years prior to onset of DM) and two sets of non-diabetic controls individually matched to cases on age, HIV status, race, ethnicity. RESULTS/ANTICIPATED RESULTS: Our metabolomic analysis detected a total of 130 identifiable metabolites with 3 metabolites differentially expressed in cases. Partial Least Squares Discriminant prediction of incident DM within the training set yielded a predictive metabolite signature enriched for amino acid and urea cycle metabolites. Pathway analysis of this predictive signature confirmed amino acid biosynthesis as significantly overrepresented. DISCUSSION/SIGNIFICANCE OF IMPACT: Our data suggest that amino acid metabolites are early markers for DM in HIV-infected and uninfected women, similar to patterns seen in the general diabetic population.
PLASMA MIR-15A AS A POTENTIAL BIOMARKER OF PANCREATIC CANCER
Ogunwobi O1, Li J1, George TJ1, Trevino J1, Liu C1 1University of Florida, Gainesville, FL, USA
OBJECTIVES/SPECIFIC AIMS: Pancreatic cancer is a very deadly cancer with overall five-year survival rate of only 6% in part due to the advanced disease stage at diagnosis. Previous research has not revealed effective non-invasive strategies for early detection and diagnosis of pancreatic cancer. Therefore, the aim of this study was to determine the potential for use of plasma miR-15a as a non-invasive biomarker for use in early detection and diagnosis of pancreatic cancer. METHODS/STUDY POPULATION: Plasma samples from patients diagnosed with pancreatic cancer at an academic medical center and plasma samples from a murine model of human pancreatic cancer were studied. Plasma samples from normal healthy subjects were used as controls. Total RNA was isolated from plasma samples and expression of plasma miR- 15a was assessed using real-time quantitative polymerase chain reaction. U6 snRNA was used as an internal control. RESULTS/ANTICIPATED RESULTS: Plasma miR-15a expression was higher in both pancreatic cancer patients and mice bearing human pancreatic cancer than normal healthy control subjects. The increased expression of plasma miR-15a was statistically significant in the human study (P = 0.022) but not in the murine study. DISCUSSION/SIGNIFICANCE OF IMPACT: This study is the first to examine the plasma expression profile of miR-15a in pancreatic cancer. Our experiments showed that plasma miR-15a expression is higher in pancreatic cancer patients and mice bearing human pancreatic cancer than normal healthy control subjects. Consequently, miR-15a may be a potential plasma biomarker for use in early detection and diagnosis in pancreatic cancer.
PREVALENCE OF PERIODONTAL DISEASE, PRE-DIABETES, AND SALIVARY CYTOKINES LEVELS AMONG OBESE ADOLESCENTS L. ORRACA AND L. LOPEZ, UNIVERSITY OF PUERTO RICO SCHOOL OF DENTAL MEDICINE, SAN JUAN, PR
Orraca LJ1 1University of Puerto Rico School of Dental Medicine, San Juan, USA
OBJECTIVES/SPECIFIC AIMS: Obesity and pre-diabetes are becoming more prevalent in children and adolescents. Published reports suggest an association of periodontal disease with obesity and pre-diabetes in adults. METHODS/STUDY POPULATION: We will perform a observational cross-sectional, population-based study of a random representative sample of 300 adolescents scholars between 13 to 18 years, in five health regions (30 obese and 30 normal per school). Trained and calibrated examiners will collect data in a questionnaire addressing obesity, nutrition, sociodemographic data and physical activity. We will perform oral examination to determine the precens of preriodontitis or gingivitis with bitewing radiographs to measure alveolar bone loss; blood test to assess glycosylated hemoglobin; salivary specimen collection for biomarker analysis; and BMI assessment. Salivary biomarkers related to PD and/or obesity will be analyzed by ELISA. Bivariable analysis to assess the 2-way associations between obesity and periodontal disease status and between the combined obesity-periodontal status variable and the levels of each of the biomarkers using Student's t-tests and Chi-square RESULTS/ANTICIPATED RESULTS: Pending DISCUSSION/SIGNIFICANCE OF IMPACT: Conduct an epidemiologic investigation that will have an impact on the oral health of the adolescent population of Puerto Rico, with potential benefit for the broader Hispanic community. If we demonstrate and association; it will permit establish new project ideas. “This work was partially supported by the NIH under Award Number R25MD007607. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.”
PROGESTERONE RECEPTOR SIGNALING INDUCES CELLULAR SENESCENCE IN OVARIAN CANCER CELLS
Diep CH1, Charles NJ1, Gilks C2, Kalloger SE2, Argenta PA1, Lange CA1 1University of Minnesota, Minneapolis, MN, USA; 2University of British Columbia, Vancouver, BC, Canada
OBJECTIVES/SPECIFIC AIMS: Define the molecular mechanisms of PR expression and its anti-tumorigenic effects in ovarian cancer cells. METHODS/STUDY POPULATION: n/a RESULTS/ANTICIPATED RESULTS: To study the suppressive role of PR in ovarian cancer in the absence of added estrogen (i.e. needed to stimulate PR expression), we created ES-2 ovarian cancer cells stably expressing vector control or GFP-tagged PR-B. Unmodified ER+/PR+ PEO4 ovarian cancer cells were included to validate our findings. Progestin stimulation (R5020; 10 nM) of ES-2 cells stably expressing GFP-PR inhibited the formation of large colonies in soft-agar assays, but yielded a significant increase in the number of viable, very small colonies relative to vehicle-treated and PR-null cohorts. Continuous treatment with R5020 induced cellular senescence characterized by altered cellular morphology, senescence-associated β-galactosidase activity, irreversible G1 cell-cycle arrest, and upregulation of the cell-cycle inhibitor, p21, as well as the Forkhead-box transcription factor, FOXO1. Notably, both PR-B and FOXO1 were detected within the same PRE-containing regions of the p21 upstream promoter. Stable knock-down using lentiviral shRNAs targeting FOXO1 inhibited progestin-induced p21 expression and blocked the development of senescence, suggesting that progestin-induced cellular senescence in PR+ ovarian cancer cells is mediated by FOXO1-dependent p21 expression. DISCUSSION/SIGNIFICANCE OF IMPACT: Overall, these findings support the concept of PR as a tumor suppressor in ovarian cancer cells that exhibits its inhibitory effects by inducing cellular senescence. Clinical targeting of the PR-FOXO1-p21 signaling pathway may provide a useful strategy to induce irreversible cell cycle arrest and sensitize ovarian cancer cells to existing chemotherapies as part of combination therapy.
PROTEOMIC PROFILE OF SERA FROM FATIGUED MEN RECEIVING LOCALIZED EXTERNAL BEAM RADIATION THERAPY FOR NON-METASTATIC PROSTATE CANCER
Lukkahatai N1, Patel S2, Gucek M2, Hsiao C1, Saligan L1 1National Institute of Nursing Research, National Institue of Health, Bethesda, MD, USA; 2National Heart, Lung and Blood Institute, National Institute of Health, Bethesda, MD, USA
OBJECTIVES/SPECIFIC AIMS: Fatigue is the most distressing side effect of radiation therapy. Fatigue severity is known to increase during the course of treatment. Etiology of fatigue progression is unknown. This study aims to examine the proteome changes from sera of fatigued men with non-metastatic prostate cancer receiving external beam radiation therapy (EBRT). METHODS/STUDY POPULATION: Fatigue scores measured using the Functional Assessment of Chronic Illness Therapy-Fatigue and serum samples were collected at baseline and midpoint of EBRT from 12 subjects enrolled in NCT00852111. Participants were grouped by the degree of change in their fatigue scores during EBRT into: high (HCF >75% quartile), moderate (MCF = 26–75% quartile), and small change in fatigue (SCF <25% quartile). Two-Dimensional Fluorescence Difference Gel Electrophoresis (2-D DIGE) and liquid chromatography-tandem mass spectrometry technology were used to analyze and identify the protein changes from the depleted sera. Western Blot analyses confirmed the protein changes. RESULTS/ANTICIPATED RESULTS: Apolipoprotein (Apo)A1, ApoE, transthyretin consistently changed (fold change > 1.5, p < 0.001) from the 2-D DIGE analyses. HCF subjects had the highest mean ApoE at baseline (mean = 2.65 ± 1.8) and at midpoint (mean = 11.88 + 9.8) of EBRT. SCF subjects had the highest transthyretin concentration at baseline (mean = 74.00 ± 28.8) and at midpoint (mean = 63.37 ± 55.7) of EBRT. MCF subjects had the highest mean ApoA1 concentration at baseline (mean = 5.56 ± 8.0) of EBRT. DISCUSSION/SIGNIFICANCE OF IMPACT: Apo E, ApoA1, and transthyretin may provide pathways that can explain the etiology behind fatigue progression in this population.
QUANTITATIVE ASSESSMENT OF THE NEONATAL T CELL RECEPTOR REPERTOIRE
Beisang D1, Rajpal M1, Georgieff M1, Jenkins M1 1University of Minnesota, Minneapolis, MN, USA
OBJECTIVES/SPECIFIC AIMS: Neonatal infection is a major cause of morbidity and mortality in the newborn intensive care unit (NICU) due to the immunocompromised state of premature infants. We hypothesize that one mechanism underlying this susceptibility to infection is a reduced diversity in the neonatal T cell repertoire due to the early developmental stage of the premature infant. METHODS/STUDY POPULATION: In order to address this question, we are measuring the diversity of the T cell receptor (TCR) beta chain (Tcrb) sequences among an antigen specific population of cells from mouse models of premature and full-term infants (mouse day of life one and seven, respectively). We are utilizing the technique of MHC Class II tetramer enrichment to purify cells specific for an immunogenic peptide 2W from mice primed with purified 2W peptide on either day of life one or seven. The Tcrb mRNA will then be sequenced using Illumina high-throughput sequencing technology and the diversity of antigen specific Tcrb sequences assessed computationally. RESULTS/ANTICIPATED RESULTS: Reads generated by sequencing will be de novo assembled and the utilization of the various V, D and J chain regions as well as untemplated sequences will be compared between the preterm and full term conditions. DISCUSSION/SIGNIFICANCE OF IMPACT: The results of these experiments will help further our understanding of the neonatal immune system as well as provide guidance for the development of more efficacious vaccines for this high-risk patient population.
RECOGNITION OF PLASMODIUM FALCIPARUM APICAL MEMBRANE ANTIGEN-1 AND CIRCUMSPOROZOITE PROTEIN SPECIFIC B-CELLS IN IMMUNIZED MICE
Frosch A1, Taylor J1, John C1 1University of Minnesota, Minneapolis, MN, USA
OBJECTIVES/SPECIFIC AIMS: Of the estimated 216 million cases of malaria worldwide annually, 655,000 of those affected will die and far more will develop disability from severe disease. Antibodies to malaria antigens have been associated with infection and disease protection, but development of durable serologic responses to even the most immunogenic malaria antigens is not reliable. Hypotheses for dysfunctional malaria specific humoral responses include B-cell exhaustion, germinal center architectural disruption, parasite antigenic variation and inhibitory innate immune signaling. No effective method for phenotypic analysis of malaria specific B cells has yet been described. Our aim in this project is to develop a method to evaluate malaria antigen specific B-cell subsets in humans. METHODS/STUDY POPULATION: We present an approach for quanitfying Plasmodium falciparum apical membrane antigen-1 (AMA-1) and circumsporozoite protein (CSP) specific B-cells in peripheral blood using tetramerized biotinylated malaria antigen coupled to phycoerythrin labeled streptavidin, magnetic bead enrichment and cytometric flow analysis. RESULTS/ANTICIPATED RESULTS: With this assay, we can detect AMA-1 and CSP specific germinal center reactions after immunization of B6 mice with P. pastoris recombinantly expressed forms of these two proteins. DISCUSSION/SIGNIFICANCE OF IMPACT: Our data suggests that this assay when applied in humans will be a valuable tool in studying naturally acquired and vaccine derived malaria specific B-cell responses.
RECOVERY OF LOCOMOTOR AND MICTURITION FUNCTION FOLLOWING SCI: TRANSLATIONAL ANIMAL AND HUMAN MODELS
Qureshi F1, Boyce V1, Collins WF1, Sisto SA1 1Stony Brook University, Stony Brook, NY, USA
OBJECTIVES/SPECIFIC AIMS: The aim is to determine if locomotor training (LT), an intervention to address the recovery potential of the spinal circuits that promote walking, and determine if this corresponds to recovery of micturition. Spinal cord injury (SCI) is a devastating condition resulting in loss of locomotor, bladder and other physiological functions. Fortunately, the brain and SC exhibit considerable plasticity. SCI animal models provide an opportunity to study central nervous system function and identify mechanisms of neuronal plasticity that could be utilized to devise rehabilitative strategies for humans. For example, studies in animal models demonstrated changes in spinal circuitry and afferent pathways that improve ambulation after LT which led to the successful application of LT to improve walking even in chronic SCI. Inversely, observations from human clinical research inform animal research where decreased walking may interrupt the micturition pathways resulting in loss of bladder control, detrusor sphincter dyssynergia and urinary tract infections. METHODS/STUDY POPULATION: SCI patients engaged in LT report improved bladder function suggesting that lumbar spinal circuitry has overlapping mechanisms for plasticity in locomotor and bladder function. While difficult to assess in humans, it is possible to test this hypothesis in a systematic manner in an animal model of SCI. Mechanisms learned in the animal can be continuously translated to SCI patients. RESULTS/ANTICIPATED RESULTS: Translational animal and human models of motor and micturition plasticity post SCI, as a result of locomotor training, will be presented. DISCUSSION/SIGNIFICANCE OF IMPACT: If there is a parallel correspondence of recovery of walking and bladder function in animals, this can be translated to the same in humans and better inform goals of urodynamic testing, prognosis and bladder management guidelines.
REGULATION OF ENDOGENOUS GLUCOSE PRODUCTION (EGP) BY CENTRAL K-ATP CHANNELS IN HUMANS IS INDEPENDENT OF BODY MASS INDEX (BMI)
Esterson YB1, Raghavan P1, Carey M1, Hawkins M1 1Albert Einstein College of Medicine, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: We reported that diazoxide, a K-ATP channel activator, significantly reduces EGP in healthy humans under fixed hormonal conditions, with parallel rat studies suggesting that these effects are centrally mediated (JCI 121: 4916, 2011). Rodent studies have suggested that central interventions are ineffective at regulating metabolism in obesity. While insulin activates central K-ATP channels in lean rats, this response was lacking in obese rats (Nat Neurosci 3:757, 2000). Similarly, while intracerebroventricular insulin reduced food intake and body weight in lean rats, these effects were not observed in obese rats (Appetite 7:381, 1986). We therefore examined whether central regulation of EGP might be impaired with obesity in humans. METHODS/STUDY POPULATION: The current study assessed the ability of diazoxide to suppress EGP in 12 nondiabetic subjects with BMI range of 20.4–33.5 kg/m2. Subjects were given oral diazoxide 4 mg/kg or placebo 3-hours prior to a 4-hour euglycemic 'pancreatic clamp' study (somatostatin infusion with basal replacement of insulin, glucagon and growth hormone). RESULTS/ANTICIPATED RESULTS: Diazoxide decreased EGP by 25% relative to placebo (1.26 ± 0.15 mg/kg/min with diazoxide vs. 1.71 ± 0.16 mg/kg/min with placebo, P < 0.005). The percent suppression of EGP did not significantly differ by BMI (normal weight subjects: -29.5 ± 19.3%; overweight and obese subjects: -24.0 ± 4.5%; p = 0.68). There was no correlation between subjects' BMI and percent suppression of EGP with diazoxide (r2 = 0.01, p = 0.73). Therefore, normal suppression of EGP by activation of K-ATP channels persisted in overweight and obese humans. DISCUSSION/SIGNIFICANCE OF IMPACT: Given the prevalence of the metabolic syndrome and its consequences, these central pathways could be pharmacologically targeted to improve glucose metabolism in high-risk individuals.
REGULATORY MARKERS PREDICT SUSCEPTIBILITY TO INFECTION IN TRAUMA PATIENTS
West SD1, Brooks T3, Ziegler A1, Mold C2 1University of New Mexico School of Medicine Department of Surgery, Albuquerque, NM, USA; 2University of New Mexico School of Medicine Department of Molecular Genetics and Microbiology, Albuquerque, NM, USA; 3University of New Mexico School of Medicine, Albuquerque, NM, USA
OBJECTIVES/SPECIFIC AIMS: After surviving the initial trauma, severely injured trauma patients become susceptible to infectious complications. This has been attributed in part to dysregulation of the immune system. Soluble CD163 (sCD163) is an anti-inflammatory marker that has been shown to be a predictor of susceptibility to sepsis in burn patients. We hypothesized that sCD163 could be used to predict susceptibility to infections in trauma patients. METHODS/STUDY POPULATION: The Human Research Review Committee approved all protocols prior to sample collection. Severely injured trauma patients defined by an Injury Severity Score of 16 or greater or requiring ICU admission were consented and blood samples were collected at enrollment and again at 48 and 72 hrs. Patients were followed through their hospital stay and any infectious complications were recorded. Soluble CD163 levels were determined in the plasma from all three blood draws by ELISA. RESULTS/ANTICIPATED RESULTS: The mean values for trauma patients on day 1 were similar to healthy controls. We found significantly decreased sCD163 on days 2 and 3 when compared to day 1 (p < 0.05) or to controls. There were significantly higher levels of sCD163 in plasma of patients who became infected during their hospital stay compared to those who did not (p < 0.05). For a 50 ng/ml increase in the amount of sCD163 from the mean, there was a 1.6 greater chance of developing an infection (p = 0.04). DISCUSSION/SIGNIFICANCE OF IMPACT: This study confirms the utility of sCD163 as a potential biomarker for infectious complications in severely injured trauma patients. Identification of at risk patients could lead to potential preventative and therapeutic interventions.
RENAL TUBULAR CELLS INDUCE ALTERNATIVE ACTIVATION IN MACROPHAGES VIA A NONCANONICAL PATHWAY
Huen SC1, Cantley LG1 1Yale University School of Medicine, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Macrophage infiltration into the kidney after ischemia/reperfusion (IR) can contribute to both the initial injury and subsequent recovery. An inflammatory phenotype predominates early after reperfusion while an alternatively activated anti-inflammatory phenotype predominates later during the repair phase. How signals from renal tubular cells influence these macrophage phenotypes is unknown. METHODS/STUDY POPULATION: Mouse proximal tubular (MPT) cells were cultured in serum free media for 48 hours to generate MPT conditioned media (CM). Bone marrow-derived macrophages (BMM) were cultured in the presence or absence of MPT CM and harvested for protein or mRNA analysis. RESULTS/ANTICIPATED RESULTS: NF-kB and Stat3 are activated in wild-type BMM cultured in MPT CM. Within 2 hours of exposure to MPT CM, macrophages express proinflammatory markers (Il-1b, Il-6). By 12 hours after MPT CM incubation, the proinflammatory genes are downregulated, whereas alternative activation markers (Arg1, MR) are upregulated. The early proinflammatory response and Arg1 was prevented by NF-kB pathway inhibitors and in MyD88-/- and TLR2-/-TLR4-/- BMM. Jak Inhibitor I inhibits both Arg1 and MR expression. The late expression of Arg1 and MR was not prevented in Il-4ra-/-, Stat6-/- or LysM-Cre;Stat3fl/fl BMM. DISCUSSION/SIGNIFICANCE OF IMPACT: Secreted tubular factors induce sequential macrophage activation from a NF-kB dependent proinflammatory phenotype to an anti-inflammatory phenotype. This sequential macrophage activation is similar to that seen in vivo after renal IR injury, suggesting that tubular cells can regulate the phenotype of infiltrating macrophages. Arg1 expression is co-regulated by a TLR/NF-kB and Jak/Stat pathway while MR expression is downstream of a Jak/Stat pathway. The alternative activation of macrophages by tubular factors occurs through a Stat3-, Stat6-independent pathway.
RESTING STATE FMRI NEURAL CORRELATES OF ECT RESPONSE IN MAJOR DEPRESSIVE DISORDER
Abbott C1, Jones TR1, Bustillo J1, Calhoun VD1,2, Turner JA1,2 1Univ New Mexico, Albuquerque, NM, USA; 2Mind Research Network, Albuquerque, NM, USA
OBJECTIVES/SPECIFIC AIMS: Electroconvulsive therapy (ECT) remains the gold-standard treatment for severe, treatment-resistant patients with major depressive disorder (MDD). The short time interval and magnitude of response make ECT an ideal therapeutic intervention to assess biomarkers of clinical improvement in MDD. Previous research has shown that MDD is associated with increased functional connectivity during resting state functional magnetic resonance imaging (fMRI) in dorsal frontal, parietal and limbic regions. Here, we hypothesized that ECT response would be associated with a reduction in functional connectivity. METHODS/STUDY POPULATION: A longitudinal design included fMRI and clinical assessments before and after the ECT series. Depressed subjects met the following inclusion criteria: 1) DSM-IV TR diagnosis of MDD; 2) the clinical indications for ECT; and 3) a Hamilton Depression Rating Scale-24 item (HDRS-24) > 21. We analyzed the preprocessed fMRI data with the CONN functional connectivity toolbox and SPM8. The average age (n = 16) was 65.63 years +/– 8.46 (4 males/11 females). The ECT series consisted of 11.81 +/– 3.35 treatments completed thrice weekly. RESULTS/ANTICIPATED RESULTS: The HDRS-24 confirmed clinical response (pre-ECT: 32.56 +/– 8.64; post-ECT: 7.56 +/– 7.82; t(15) = 8.28, p < 0.001). Paired t-tests revealed significant reductions in functional connectivity in the right dorsal medial frontal cortex (extent threshold of FWE-corrected p = 0.011; height threshold p < 0.001). DISCUSSION/SIGNIFICANCE OF IMPACT: The diminished functional connectivity of the dorsal medial prefrontal cortex may be related to restoration of emotional regulation of patients that respond to ECT. Reduced functional connectivity of dorsal frontal cortical regions may be an important biomarker of response in MDD.
ROLE OF MEMBRANE-TYPE 1 MATRIX METALLOPROTEINASE IN THE PATHOGENESIS OF THORACIC AORTIC ANEURYSMS
Ward AF1, Mignatti P1, Grossi EA1, Ursomanno P1, Maddalo S1, DeAnda A1 1NYU School of Medicine, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: Background: The histopathologic hallmark of thoracic aortic aneurysms (TAAs) is medial degeneration with extensive loss of elastic lamellar proteins, including fibrillin-1 (Fbn-1). TAA in Marfan Syndrome is associated with Fbn-1 mutations. Fbn-1 is degraded by membrane-type 1 matrix metalloproteinase (MT1-MMP) in vitro. While elevated levels of MT1-MMP have been associated with TAA, its effect on Fbn-1 in TAA is unknown. Objectives: We attempted to characterize tissue MT1-MMP and Fbn-1 in normal and TAA patients (non-Marfan) to determine if Fbn-1 degradation is correlated with elevated MT1-MMP levels. METHODS/STUDY POPULATION: MT1-MMP and Fbn-1 levels were measured in TAA tissue from patients undergoing repair and in control aorta (CON) from patients undergoing coronary artery bypass grafting. Specimens were homogenized, protein was extracted and quantified, and the extracts were analyzed by Western blotting with antibodies to human MT1-MMP and Fbn-1. RESULTS/ANTICIPATED RESULTS: Elevated levels of MT1-MMP were detected in 11/13 TAA, while 4/6 CON had undetectable levels (p < 0.02). In preliminary subset analysis, Fbn-1 degradation products were detected in 2 TAA samples with high levels of MT1-MMP but no detectable Fbn-1 degradation products were found in 2 CON samples that showed low levels of MT1-MMP. DISCUSSION/SIGNIFICANCE OF IMPACT: Elevated levels of MT1-MMP were associated with Fbn-1 degradation in TAA. This mechanism may contribute to an increased susceptibility to the development of TAA and may serve as a future therapeutic target for the detection and management of TAA.
ROLE OF MET AMPLIFICATION IN ALTERING THE DNA DAMAGE RESPONSE (DDR) IN GLIOBLASTOMA (GBM)
Gillam M1, Camacho C1, Tomimatsu N1, Todorova P1, Gil del Alcazar C1, Mukherjee B1, Burma S1 1University of Texas Southwestern Medical Center, Dallas, TX, USA
OBJECTIVES/SPECIFIC AIMS: The overarching goal of this study is to understand in GBM whether genomic instability generated during radiation therapy might promote the development of tumor sub-populations with MET amplification that contribute to therapy resistance via enhancement of the DDR. METHODS/STUDY POPULATION: Transgenic mouse models with brain-targeted deletions of GBM-relevant tumor suppressors were utilized to study ionizing radiation (IR)-induced gliomagenesis. Mice were irradiated with X-rays or Fe ions and then monitored for tumor formation. Radiation-induced GBMs were characterized by array CGH and FISH. To understand the role of the most significant genetic alteration observed, MET amplification, ex vivo cultures with MET expression were analyzed for stem cell properties, tumorigenic potential, DDR protein levels, and IR-induced G2/M checkpoint activity. RESULTS/ANTICIPATED RESULTS: Characterization of the IR-induced tumors revealed striking amplification of the MET oncogene. The MET-expressing cells display cancer stem cell (CSC)-like characteristics, namely, neurosphere formation in stem cell media and expression of Sox2. MET expression also augments the tumorigenic potential of these cells. Importantly, high levels of MET resulted in upregulation of numerous DDR proteins and induced a more durable cell cycle checkpoint response. DISCUSSION/SIGNIFICANCE OF IMPACT: MET expression has been shown to support a CSC phenotype in GBM. Furthermore, recent evidence suggests that GBM CSCs are radioresistant because of an enhanced DDR, the mechanistic basis of which is unknown. We postulate that induction of DDR proteins by MET might underlie therapeutic radioresistance of GBM CSCs. Understanding the mechanism by which MET augments DDR will aid in the development of strategies to sensitize these tumors with MET inhibitors.
ROLE OF SPHINGOLIPID SIGNALLING IN BRONCHOPULMONARY DYSPLASIA
Harijith A1, Reddy N1, Bai T1, Usatyuk P1, Pendyala S1, Huang L1, Berdyshev E1, Gorshkova I1, Reddy S1, Raj U1, Natarajan V1 1University of Illinois, Chicago, IL, USA
OBJECTIVES/SPECIFIC AIMS: Bronchopulmonary Dysplasia (BPD) of the premature newborn is characterized by lung injury resulting in alveolar simplification and reduced pulmonary function. We have earlier shown that exposure of adult mice to hyperoxia enhances sphingosine-1-phosphate levels in lung tissues; however, the role of increased S1P generation and signaling in the pathobiology of BPD has not been investigated. We hypothesized that altered S1P signaling axis in BPD, in part, is responsible for neonatal lung injury and inflammation. METHODS/STUDY POPULATION: To test this, newborn (NB) C57BL6/J, SphK1-/-, SphK2-/- and Sgpl1+/- (S1P lyase) mice were exposed to hyperoxia (75%) from postnatal day1 to 7, pups were sacrificed on postnatal day 7, and lungs were perfused and used for histology and biochemical analysis. RESULTS/ANTICIPATED RESULTS: SphK1-/-, but not SphK2-/-, deficiency in mice offered protection against hyperoxia-induced lung injury compared to wild type littermate controls. Histopathology revealed better formed alveoli and comparable levels of proteins in bronchoalveolar lavage (BAL) fluids in SphK1 deficient mice. Further, hyperoxia increased levels of pro-inflammatory IL-6 was decreased in bronchoalveolar lavage fluids in SphK1 deficient mice compared to the wild type. Analysis of total cell lysates by Western blots showed decreased expression of Nox-2 and Nox-4 proteins, in the lung tissue of SphK1-/- mice exposed to hyperoxia compared to its wild type control. DISCUSSION/SIGNIFICANCE OF IMPACT: These results suggest an important role for SphK1 and S1P signaling in the development of hyperoxia-induced lung injury in a murine neonatal model of BPD. Funding (in part) by the National Center for Advancing Translational Sciences, National Institutes of Health, through Grant UL1TR000050.
S100A8/A9 REDUCES HYPOXIA INDUCIBLE FACTOR AND TUMOR GROWTH IN HEAD AND NECK SQUAMOUS CELL CARCINOMA
Sorenson BS1, Gopalakrishnan R1, Herzberg MC1 1University of Minnesota, Minneapolis, MN, USA
OBJECTIVES/SPECIFIC AIMS: Head and neck squamous carcinomas (HNSCC) have poor outcomes with standard radiation and chemotherapy. Anti-tumor therapies may be subverted by low oxygen tension, where Hypoxia Inducible Factors (HIFs) can be induced and promote tumor growth and cellular survival. During low oxygen tension, the protein complex S100A8/A9 down-regulates HIFs as we have shown. We now seek to learn whether forced expression of S100A8/A9 in vivo down-regulates HIFs and suppresses tumor growth. METHODS/STUDY POPULATION: S100A8/A9 positive and negative carcinoma cell lines were evaluated in vitro for expression of HIF proteins in response to low oxygen tension. The cell lines were then inoculated subcutaneously in immunodeficient mice and evaluated for tumor growth. Future in vivo studies will focus on tumor-delivered S100A8/A9 mRNAs for altered HIF expression and anti-tumor activity. RESULTS/ANTICIPATED RESULTS: In response to low oxygen tension in vitro, S100A8/A9 expression decreased HIFs and proinflammatory cytokines. In vivo, ectopic expression of S100A8/A9 significantly reduced formation of new tumors. In planned studies, we expect to show that intra-tumor delivery of S100A8/A9 mRNAs will suppress tumor growth and reduce tumor volume in vivo. DISCUSSION/SIGNIFICANCE OF IMPACT: If our hypothesis is confirmed, these data will show that intra-tumor delivery of S100A8/A9-specific mRNAs may be a novel adjunctive therapy for HNSCC. By down-regulating HIFs, S100A8/A9-specific mRNAs are expected to reduce tumor subversion of conventional anticancer therapies. The effective dose of radiation and chemotherapeutics may be reduced, minimizing adverse events and increasing the frequency of successful therapy.
SERUM MMP-3 AND MMP-7 PREDICT LUNG INJURY IN NYC FIREFIGHTERS
Kwon S1, Cho SJ1, Naveed B1, Comfort A1, Prezant DJ2,3, Rom WN1, Weiden MD1,2, Nolan A1 1NYU School of Medicine, New York, NY, USA; 2Fire Department of New York, Brooklyn, NY, USA; 3Montefiore Medical Center, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: After 9/11/2001, some FDNY workers had excessive lung function decline whereas other similarly exposed workers were unaffected. Workers with progressive disease, termed World Trade Center Lung Injury (WTC-LI), had bronchial wall thickening and air trapping on CT, suggesting airway injury. MMPs have been studied for their role in tissue and lung injury. We hypothesized that serum levels of MMPs present within months of 9/11 can predict sensitivity to airway injury years later. METHODS/STUDY POPULATION: This was a case-cohort analysis of N = 801 never-smoking firefighters who had: normal pre-9/11 FEV1, serum biomarkers drawn at Medical Monitoring Entry (MME) prior to 2/11/2002, and had FEV1 measurements at subspecialty pulmonary evaluation (SPE). SPE was 32 months after 9/11. We assayed MMP-1,2,3,7,8,9,12,13 in serum collected at MME. A randomly selected control sub-cohort of N = 137/801 was drawn from the N = 801 population. Cases with WTC-LI were those who had the lowest FEV1 values of the N = 801 population. Logistic regression was used to develop a prediction model of developing WTC-LI using biomarker cutpoints. RESULTS/ANTICIPATED RESULTS: In cases, median FEV1 consistently declined from pre-9/11 to MME to SPE (88% to 77% to 72%). In controls, FEV1 initially declined, and then recovered (102% to 91% to 95%). In a multi-analyte model adjusted for pre-9/11 FEV1, BMI at SPE, age and exposure, having MMP-3 < 3.1 ng/mL increased odds of developing WTC-LI 410%; having MMP-7 ≥ 354.84 pg/mL increased odds 271%. DISCUSSION/SIGNIFICANCE OF IMPACT: Decreased levels of MMP-3 and elevated levels of MMP-7 present in serum early after 9/11/2001 increased the likelihood of WTC-LI compared to controls. MMP-3 and MMP-7 reflect independent pathways to airway injury and repair, and can predict future lung injury.
SLEEP, GENE EXPRESSION AND CHRONIC GI SYMPTOMS
Henderson WA1, Rasmussen N1, Berger R1, Wang X1, Martino A1, Reddy S1, Longchamps R1, Fourie N1, Heitkemper MM2 1NIH, NINR, DHHS, Bethesda, MD, USA; 2University of Washington, Seattle, WA, USA
OBJECTIVES/SPECIFIC AIMS: Persons with chronic gastrointestinal (GI) symptoms such as chronic abdominal pain (CAP) often have poor sleep quality. Brain-derived neurotrophic factor (BDNF) has been shown to plan a pivotal role in many pain conditions. This study explored the association between sleep quality, gene expression and BDNF genotype in persons with and without CAP. METHODS/STUDY POPULATION: The sample (n = 59, 45.8% male, 61% Caucasian, 26.9 ± 6.64 years) included 19 consented persons with CAP and 40 controls. Sleep quality was measured by the Pittsburgh Sleep Quality Index (PSQI) and global scores ≥5 were categorized as poor sleep quality. CAP (>6 months) was determined by both self-report and medical record review. RNA extracted from whole blood was used for gene expression via GeneChip Human Genome U133+2.0 array (containing 47,000 transcripts); BDNF genotyping was completed in a race-matched subset (n = 26). RESULTS/ANTICIPATED RESULTS: Persons with CAP had significantly higher PSQI scores (p = 0.006), sleep latency (p = 0.014), sleep disturbances (p = 0.046), and daytime dysfunction (p = 0.006) than controls. In the matched subset, participants with CAP and poor sleep were equally homozygous and heterozygous for BDNF single-nucleotide polymorphism. Clustering analysis of the interaction of BDNF and gene expression revealed of insulin-like growth factor 1 (IGF1), spermatogenesis associated serine-rich 2-like (SPATS2L) and immunoglobulin heavy constant gamma 1 or mu (IGHG1) were significantly dysregulated in the those with comorbid poor sleep quality and CAP. DISCUSSION/SIGNIFICANCE OF IMPACT: Our findings support the association of poor sleep quality with GI symptoms. Furthermore, the additive effect of BDNF genotype together with gene expression and associated pathways may have pivotal regulatory relationships in patients with comorbid sleep and GI symptoms.
SYSTEMIC MICROVASCULAR DYSFUNCTION IN PULMONARY ARTERIAL HYPERTENSION
Tonelli A1, Cikach F1, Dababneh L1, Newman J1, Dweik R1 1Cleveland Clinic, Cleveland, OH, USA
OBJECTIVES/SPECIFIC AIMS: Pulmonary arterial hypertension (PAH) is a condition characterized by elevated pulmonary pressures that can lead to right heart failure and death. These patients have pulmonary vascular involvement; however it is unclear whether they also have systemic vascular dysfunction. We tested the hypothesis that patients with PAH have systemic (not only pulmonary) vascular dysfunction. METHODS/STUDY POPULATION: The study received IRB approval and informed consent was obtained. Right heart catheterization (RHC) was performed for confirmation of PAH diagnosis or a part of follow-up of the condition. Immediately after RHC we studied the systemic vasculature using skin blood flow dynamic responses to temperature and arm occlusion. Forearm skin perfusion was determined by laser Doppler fluxmetry (Periflux system 5000, Perimed, Sweden) using perfusion units (PU). RESULTS/ANTICIPATED RESULTS: We included 23 adult patients with PAH ((idiopathic: 10, portopulmonary: 5, associated with connective tissue disease: 6, HIV: 1 and congenital heart disease: 1) and 8 individuals without pulmonary hypertension (PH). Baseline skin perfusion was higher in patients with PAH (9.9 ± 4.8 PU) vs no PH (6.3 ± 2.5 PU), p = 0.01. Thermal hyperemia percentage change had a trend to be higher in patients without PH (2916 ± 2362%) than those with PAH (1966 ± 1167%), p = 0.1. Postocclusive reactive hyperemia percentage change was higher in PAH patients (916 ± 720%) than those without PH (548 ± 189%), p = 0.05. Baseline PU was associated with pulmonary vascular resistance (R = –0.4, p = 0.04) and time to peak flow with cardiac output (R = 0.67, p = 0.05). DISCUSSION/SIGNIFICANCE OF IMPACT: In this proof-of-concept study, we showed that patients with PAH have systemic microvascular dysfunction compared with patients without PH.
TELOMERE LENGTH AND FEMALE REPRODUCTIVE OUTCOMES
Kalmbach K1, Antunes D1, Seth-Smith M1, Wang F1, Liu L1, Keefe D1 1New York University Medical Center, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: Fecundity begins a dramatic decline by the third decade of life. By age 40, over half of women are overtly infertile, despite an average age of menopause onset in the early 50s. In mouse models, experimental shortening of telomeres recapitulates the phenotype of reproductive aging in women, including reduced synapsis and chiasmata, increased embryo fragmentation, and chromosome abnormalities. In humans, shorter-than-average telomere length has been associated with mothers of Down's Syndrome neonates and recurrent miscarriage. Here, we prospectively evaluate telomere length (TL) in female reproductive outcomes in an infertile population. METHODS/STUDY POPULATION: Discard eggs from women seeking fertility treatment at the New York University Fertility Center were assayed for TL by real-time polymerase chain reaction (PCR). TL was compared between women who achieved successful pregnancy and those that did not within one IVF cycle. RESULTS/ANTICIPATED RESULTS: Telomere length was positively associated with favorable reproductive outcomes after controlling for age, male factor, and tubal defects. DISCUSSION/SIGNIFICANCE OF IMPACT: Telomere length dynamics in the germline differ between men and women and across the lifespan of individuals. Here, we provide the most direct evidence of the limiting role of telomere length in the female germline.
TENOFOVIR, A POTENT ANTI-VIRAL AGENT, IS A CD73 INHIBITOR THAT PREVENTS DERMAL FIBROSIS IN A MURINE MODEL OF SCLERODERMA
Feig JL1, Tivon D1, Perez-Aso M1, Cardozo T1, Cronstein BN1 1New York University School of Medicine, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: The antiviral agents adefovir and tenofovir are AMP analogues that resemble substrates of CD73. We previously reported that adenosine, generated by the CD73-mediated dephosphorylation of AMP, plays a critical role in development of both hepatic and dermal fibrosis in murine models of cirrhosis and scleroderma, respectively. A recent clinical trial demonstrated that tenofovir, but not other antiviral agents, reverses hepatic fibrosis/cirrhosis in patients with hepatitis B. We proposed that tenofovir's antifibrotic effects are mediated by inhibition of adenosine production by CD73-mediated dephosphorylation of AMP. METHODS/STUDY POPULATION: In silico modeling used an ICM-Browser. CD73 activity was quantitated by malachite green. Bleomycin (0.25 U, SubQ)-treated mice were treated with vehicle, Adefovir, or Tenofovir (75 mg/kg, IP) [n = 5 per group]. Breaking tension was measured using a tensiometer. Hydroxyproline was quantified by colorimetric assay. Adenosine was analyzed with HPLC. RESULTS/ANTICIPATED RESULTS: In silico modeling data suggested that both adefovir and tenofovir bound to the enzymatic pocket of CD73. Tenofovir, but not adefovir, inhibited CD73 activity of 293T cells overexpressing CD73 and of recombinant enzyme. Tenofovir significantly decreased adenosine levels in the skin of bleomycin-challenged mice. Tenofovir diminished bleomycin-induced dermal fibrosis in bleomycin-treated mice (reduction of dermal thickness, breaking tension, and hydroxyproline content). Picrosirius red staining showed diminished dense collagen fibrils in tenofovir-treated mice. DISCUSSION/SIGNIFICANCE OF IMPACT: Tenofovir reduces fibrosis via inhibition of adenosine production from AMP by CD73. Tenofovir may have therapeutic potential in treating fibrosis in patients suffering from non-viral fibrosing diseases such as scleroderma.
THE IMPACT OF CONTINUOUS VERSUS INTERMITTENT HIGH FREQUENCY STIMULATION OF THE NUCLEUS ACCUMBENS ON ETHANOL PREFERENCE AND CIRCADIAN LOCOMOTOR ACTIVITY IN ALCOHOL PREFERRING RATS
Abulseoud OA1, Ruby CL1, Kale R1, Ahmed A1, Tye S1,2, Kouzani A2, Lee K1, Frye M1, Choi D1 1Mayo Clinic, Rochester, MN, USA; 2Deakin University, Melbourne, VIC, Australia
OBJECTIVES/SPECIFIC AIMS: This study aimed to investigate the effect of different patterns of high frequency stimulation at the nucleus accumbens on ethanol preference and circadian locomotor activity in adult male alcohol preferring (P) and non-preferring (NP) rats. METHODS/STUDY POPULATION: High frequency stimulation (amp = 1 V, frequency = 130 Hz, pulse width = 0.1 ms)) in the right NAc (Coordinates from bregma: AP = +2.0 mm, ML = ± 1.2 mm, DV = –6.5 mm from dura) was initiated at one of three stimulation patterns: (1) Continuous stimulation for 7 days using a small battery powered device (n = 6), (2) intermittent stimulation for 4 hours initiated at the beginning of the light phase (n = 6), or (3) intermittent stimulation for 4 hours initiated at the beginning of the dark phase using the MINCS device (n = 7). Ethanol and water intake were monitored for at least a week after stimulation. Circadian locomotor activity counts in home cage were recorded using an infrared motion detector interfaced with a computerized data acquisition system (Clocklab) and later analyzed using MatLab software. RESULTS/ANTICIPATED RESULTS: Continuous stimulation of the NAc reduced ethanol consumption and preference in P rats (table 1), with little effect on either parameter in NP rats. In contrast, NAc stimulation only during the dark phase or light phase induced circadian arrhythmia in P rats with a concomitant increase in ethanol intake noted during nighttime stimulation. DISCUSSION/SIGNIFICANCE OF IMPACT: These results suggest that alignment of circadian rhythms by continuous NAc stimulation may play a role in reducing drinking in alcohol preferring rodents. Further investigation for the impact of HF stimulation on NAc CLOCK gene expression is warrented.
THE PHARMACOGENOMIC CONTROL OF CLOPIDOGREL RESPONSE IN ACUTE CORONARY SYNDROMES
Scott SA1, Collet J2, Yang Y1, Peter I1, Shuldiner AR3, Montalescot G2, Desnick RJ1, Hulot J1,2 1Mount Sinai School of Medicine, New York, NY, USA; 2Pitié-Salpêtrière University Hospital, Paris, France; 3University of Maryland School of Medicine, Baltimore, MD, USA
OBJECTIVES/SPECIFIC AIMS: To identify genetic variants in additional to known CYP2C19 alleles associated with dual antiplatelet therapy (DAPT; clopidogrel and aspirin) response variability. METHODS/STUDY POPULATION: Candidate genes (e.g., PEAR1) recently associated with platelet aggregation and aspirin and/or clopidogrel response have been genotyped in the CLOVIS2 (NCT00822666) clinical trial participants. In addition, a genome-wide approach to identifying novel genetic variants associated with clopidogrel response variability has been undertaken to identify novel genes and variants associated with DAPT response variability. RESULTS/ANTICIPATED RESULTS: Initial PEAR1 analyses did not identify significant associations between the tested variants and clopidogrel pharmacodynamics. However, whole-exome sequencing of a cohort of carefully selected extreme phenotype DAPT-treated ACS/PCI patients has identified candidate genes and variants that are currently being validated using both the CLOVIS2 and ARCTIC (NCT00827411) clinical trial subjects. DISCUSSION/SIGNIFICANCE OF IMPACT: These results could facilitate the adoption of genetically guided antiplatelet therapy into routine interventional cardiology practice for more personalized and effective care. The identification of additional genes and variants will not only influence the uptake of personalized antiplatelet therapy, but could provide a benchmark for forthcoming pharmacogenomic studies on other drugs and medications with variable responses.
THE ROLE OF MXD3 IN HUMAN PRECURSOR B CELL ACUTE LYMPHOBLASTIC LEUKEMIA
Satake N1, Barisone G1, Lewis C1, Lam K1, Nolta J1, Diaz E1 1UC Davis, Sacramento, CA, USA
OBJECTIVES/SPECIFIC AIMS: MXD3 is a transcription factor previously shown in our lab to be a novel member of the Hedgehog pathway. The objective of this study was to investigate the role of MXD3 in precursor B (preB) ALL cell proliferation. METHODS/STUDY POPULATION: Leukemia samples were obtained either from pediatric patients or mouse xenograft models established by Satake. mRNA was measured by qRT-PCR. Reh (cell line) cells were transduced with lentiviruses carrying an shRNA sequence, either to MXD3 or the negative control, plus EGFP. Transduction efficiency was monitored by flow cytometry. Knockdown was confirmed by immunoblot with an anti-MXD3 antibody. RESULTS/ANTICIPATED RESULTS: We observed 13 to 35 times higher levels of MXD3 expression in 8 primary preB ALL samples, as well as in the preB ALL cell lines Reh and JM1, than in mobilized peripheral blood mononuclear cells from healthy donors. We investigated the role of MXD3 in cell proliferation and survival by silencing MXD3 in the Reh cell line. Upon transduction by the viral vector, MXD3 knockdown was confirmed at both the RNA and protein level. Within 48 hours, MXD3 protein levels were reduced >90% in cells infected with the shMXD3 virus but not with the control virus. To understand the role of MXD3 in leukemia cells, we analyzed cell cycle progression and apoptosis levels after knockdown using flow cytometry. We observed no significant differences in the G0/G1, S or G2/M populations between experimental and control samples. However, samples where MXD3 had been knocked down showed higher levels of apoptosis when compared to controls. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results suggest that MXD3 is important for preB ALL cell proliferation, possibly by acting as an anti-apoptotic factor. Therefore, MXD3 may represent a suitable candidate for future efforts in developing targeted therapies against preB ALL.
TRANSIENT REGULATORY T-CELL (TREG) DEPLETION WITH IL-2 DIPHTHERIA TOXIN FUSION PROTEIN (DD) ENHANCES CLEARANCE OF ACUTE MYELOID LEUKEMIA (AML) BY HAPLOIDENTICAL NATURAL KILLER (NK) CELLS
Bachanova V1, Cooley S1, Defor T1, Blazar B1, Hippen K1, McKenna D1, Weisdorf D1, Miller J1 1University of Minnesota, Minnespolis, MN, USA
OBJECTIVES/SPECIFIC AIMS: Adoptive transfer of haploidentical NK cells can induce remissions in AML. Failure may be in part a result of suppression from host Treg. We aim to elucidate the human Treg and NK cell interactions with translation of T reg depletion strategy to clinic. METHODS/STUDY POPULATION: Effect of IL-2 or IL-15 ± T reg on NK cell proliferation was measured by CFSA dilution and IL-2 in supernatant by ELISA. Next, we incorporated DD to deplete T reg and enhance NK cell expansion after adoptive transfer. Patients with refractory AML (n = 15) and non-Hodgkin lymphoma (NHL; n = 6) received chemotherapy with DD, haploidentical NK cells and IL-2. RESULTS/ANTICIPATED RESULTS: In vitro, NK cell proliferation was potently inhibited by Treg with low dose Il-2 but not with high dose IL-2. IL-2 exhaustion in supernatant suggested that Treg competed for IL-2. With IL-15, NK cell proliferation inhibition by Treg was ≤10%. In AML clinical trial, donor DNA at day 7 was detectable in 10 patients (75%; median chimerism 49%) and correlated with leukemia clearance (p = 0.05). At day 14, 28% AML patients successfully expanded NK cells and 64% attained complete remission. Less 5% Treg in blood (n = 6) at day 7 correlated with donor NK cell expansion. In contrast, > 10% Treg (n = 9) was associated with lack of donor NK expansion. NHL patients failed to expanded NK cells and 1 achieved remission. DISCUSSION/SIGNIFICANCE OF IMPACT: We showed an increased rate of donor NK cell expansion and AML remissions after a single dose of DD, but IL-2 induced host T reg rebound was not overcome in all patients. Lack of expansion in NHL patients suggests that the host/cancer milieu impact immune interactions. Novel T reg depletion methods and the use of IL-15 to expand NK cells are planned.
TUMOR SUPPRESSOR CDKN3 CONTROLS MITOSIS
Nalepa G1, Barnholtz-Sloan J2, Enzor R1, Dey D1, Gehlhausen JR1, Lehmann AS1, He Y1, Yang F1, Parada LF3, Clapp W1 1Indiana University School of Medicine, Indianapolis, IN, USA; 2Case Western Reserve University Comprehensive Cancer Center, Cleveland, OH, USA; 3University of Texas Southwestern Medical Center, Dallas, TX, USA
OBJECTIVES/SPECIFIC AIMS: Mitosis is controlled by a network of kinases and phosphatases. We screened a library of siRNAs against a genome-wide set of phosphatases to comprehensively evaluate the role of human phosphatases in mitosis. METHODS/STUDY POPULATION: n/a RESULTS/ANTICIPATED RESULTS: We discovered four candidate spindle checkpoint phosphatases, including the tumor suppressor CDKN3. We show that CDKN3 is essential for normal mitosis and G1/S transition. We demonstrate that subcellular localization of CDKN3 dynamically changes throughout the cell cycle. We show that CDKN3 dephosphorylates threonine-161 of CDC2 during mitotic exit and we visualize CDC2[pThr-161] at kinetochores and centrosomes in early mitosis. We performed a phosphokinome-wide mass spectrometry screen to find effectors of the CDKN3-CDC2 signaling axis. We found that one of the identified downstream phosphotargets, CKβ phosphorylated at serine-209, localizes to mitotic centrosomes and controls the spindle checkpoint. Finally, we show that CDKN3 protein is downregulated in glioblastoma brain tumors. DISCUSSION/SIGNIFICANCE OF IMPACT: We discovered a role for the CDKN3 tumor suppressor in human spindle assembly checkpoint. Our evidence indicates that loss of CDKN3 leads to disrupted CDC2[pThr-161] dephosphorylation in late mitosis and delayed inactivation of this key CDK during the mitotic exit. We postulate that CDKN3-deficient cells are at risk of genomic instability and transformation due to the weakened spindle checkpoint. The identification of abnormal activity of the CDKN3-CDC2 axis in glioblastoma provides opportunities for experimental therapeutics in glioblastoma and other malignancies.
VARIABLE RECOVERY OF THE MAJOR PHYLA IN THE GUT MICROBIOME AFTER COLONOSCOPY BOWEL PREPARATION
Cho I1 1New York University School of Medicine, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: Current studies of the gut microbiome frequently utilize microbial DNA extracted from stool. However, there are few studies evaluating the effect of a polyethylene glycol bowel preparation on the colonic microbiome and its subsequent recovery. Study aim: To study the longitudinal effects of polyethylene glycol bowel preparation on the taxonomic characteristics of the gut microbiome. METHODS/STUDY POPULATION: Nine patients undergoing screening colonoscopies were enrolled into this study. All patients provided a pre-colonoscopy stool sample at enrollment. Samples were collected immediately after bowel preparation with polyethylene glycol and then at weekly intervals for four weeks after the procedure. Bacterial genomic DNA was extracted from each sample. Quantitative PCR was performed targeting 16S rRNA and specific for total Bacteria, Firmicutes, and Bacteroidetes. RESULTS/ANTICIPATED RESULTS: Total Bacteria were significantly decreased immediately after bowel preparation (5.39 × 10^4 ± 1.59 × 10^4 copies/μl) when compared to baseline counts (2.98 × 10^5 ± 4.96 × 10^4 copies/μl, p < 0.001) and returned to baseline by post-procedure week 2. Firmicutes and Bacteroidetes both significantly decreased immediately after bowel preparation when compared to baseline but recovered at different rates (post-procedures weeks 1 and 4, respectively). DISCUSSION/SIGNIFICANCE OF IMPACT: Polyethylene glycol bowel preparation significantly alters the colonic microbiome immediately after the preparation. Although total Bacterial counts return to baseline levels by post-procedure week 2, the rates at which the major constituent phyla of the gut microbiome recover vary significantly. These data suggest that exposure to a bowel preparation regimen be considered when pursuing future studies in the gut microbiome.
WHOLE BLOOD GENE EXPRESSION TO STUDY PATHOGENESIS OF EARLY SEPSIS-RELATED ACUTE LUNG INJURY
Kangelaris KN1, Stein J1, Liu KD1, Aouizerat B1, Woodruff PG1, Matthay MA1, Calfee CS1 1University of California, San Francisco, San Francisco, CA, USA
OBJECTIVES/SPECIFIC AIMS: To utilize whole genome expression to identify candidate genes of early sepsis-related acute lung injury. METHODS/STUDY POPULATION: We collected whole blood RNA in critically ill patients admitted from the Emergency Department (ED) to the ICU within 24 hours of presentation at a tertiary care center. Whole genome expression was measured using the Affymetrix Human GeneChip® Gene 1.0 ST array in 75 patients with consensus-defined sepsis +/- AECC-defined ALI. The “Robust Multi-Array” (Affymetrix) was used for normalization of array values on a log2 scale. We combined a candidate gene approach with a whole genome analysis and considered genes with both > 1 log2 fold change and a false-discovery rate (FDR, Benjamini 1995) of <0.25 to correct for multiple comparisons. RESULTS/ANTICIPATED RESULTS: First, in a feasibility pilot of 12 patients with sepsis and 12 patients with sepsis + ALI, we identified several genes with > 1 log2 fold change that had been previously described in the literature in association with ALI, although the FDRs were not statistically significant (FDR >0.25). We are now measuring the gene expression of whole blood RNA in 51 additional patients with sepsis +/-ALI and will determine whether similar genes meet statistical significance in a larger sample. DISCUSSION/SIGNIFICANCE OF IMPACT: These preliminary results suggest that identification of biologically plausible genes involved in ALI pathogenesis early in the course of illness using whole blood gene expression is likely.
WHOLE EXOME SEQUENCING IN FAMILY WITH UNSPECIFIED GENODERMATOSIS OF SCLERODERMOID FEATURES
Huynh T1, Theos A2, Korf BR3,1, McKay K2, Crowley M3, Crossman D3, Sami N2 1University of Alabama Birmingham, Birmingham, AL, USA; 2University of Alabama Birmingham - Dermatology, Birmingham, AL, USA; 3University of Alabama Birmingham - Genetics, Birmingham, AL, USA
OBJECTIVES/SPECIFIC AIMS: Genodermatoses consist of inherited dermatologic disorders with associated single-gene mutation. We have identified a family with unique sclerodermoid features. Diagnosis has remained inconclusive due to mixture of clinical features consistent for scleroderma, palmoplantar keratoderma (PPK), or Huriez. Our objective is to elucidate genetic mutation associated with this undiagnosed genodermatosis. METHODS/STUDY POPULATION: Informed consent was obtained from all subjects and approved by IRB. History, physical exam, and family pedigree were performed (n = 4). Skin biopsies were taken from affected members (n = 3). Genomic DNA was isolated from peripheral blood samples. Next generation sequencing (NGS) with exome capture is conducted prior to data processing, variant discovery, genotyping, and integrative analysis. RESULTS/ANTICIPATED RESULTS: Affected members had varying degrees of hyperpigmented macules on acral surfaces; firm indurated plaques around mouth and distal fingers; and erythematous, scaly plaques with some fissuring on bilateral hands and feet. Calcinosis, Raynaud's phenomenon, sclerodactyly, and dysphagia were exhibited. Biopsies showed stratum corneum with hyperorthokeratosis and alternating parakeratosis. Keratinocytes in spinous layers had eosinophilic inclusions. We anticipate finding a novel genetic mutation; however, possible mutations include FBN1 15q21.1 for familial scleroderma or AAGAB 15q22 for PPK. DISCUSSION/SIGNIFICANCE OF IMPACT: Exome sequencing has the potential to provide diagnostics and genetic pathology for uncharacterized genodermatosis. We hope to demonstrate that NGS can be used as an initial, informative approach to study rare inherited skin diseases.
Research Professionals Abstracts Best Practices
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- SCHOLAR/TRAINEE ABSTRACTS
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
- Research Professionals Abstracts Best Practices
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
ASSESSING A SMALL GROUP MENTORING APPROACH TO ENHANCE CAREER DEVELOPMENT (K) APPLICATIONS
Anderson L1, Bolz SL1, LaPensee E1, Havermahl T1, Clauw D1 1University of Michigan, Ann Arbor, MI, USA
OBJECTIVES/SPECIFIC AIMS: Obtaining a career development (K) award (CDA) is a critical step in the career path to independence, as K scholars are more likely to earn independent funding in the future. The NIH success rate for CDAs ranged from 22% to 42% in 2011. We evaluated a three-session interactive workshop called the K Writing Series, developed to maximize support for investigators writing a CDA. METHODS/STUDY POPULATION: Between December 2009 and January 2012 we hosted 3 K Writing Series workshops. 99 junior investigators from 8 non-medical school colleges and 18 departments in the School of Medicine have attended. Activities include e-learning modules, in-person presentation, exchange and critique of draft sections, and mentor participation. RESULTS/ANTICIPATED RESULTS: Participants remark in surveys that they value the exchange of draft proposals and the guidance from peers and study section experts. To assess the effectiveness of the K Writing Series, we will compare the success rate for all K writing workshop attendees (from any school at the university) compared to the overall success rate for the School of Medicine. Additionally, frequency of workshop attendance (1–3 times) will be analyzed for those who were successful, to determine if a participants who attended more workshop sessions were more likely to get funded. DISCUSSION/SIGNIFICANCE OF IMPACT: This small group mentoring approach fosters peer critique and exchange of ideas among an interdisciplinary group of K writers. Participants are highly competitive applicants and group mentoring maximizes resources.
FEASIBILITY AND ACCEPTABILITY OF A RESEARCH OSCE FOR ASSESSMENT OF STUDENT COMPETENCE IN CTSA-DEFINED DOMAINS
Eakin B1, Lantz M1, Guenther M2, Whitman L2 1University of Michigan - MICHR, Ann Arbor, MI, USA; 2University of Michigan School of Dentistry, Ann Arbor, MI, USA
OBJECTIVES/SPECIFIC AIMS: OSCEs (Objectively Structured Clinical Examination) have been used extensively in health professions education and licensure examinations to assess student/graduate competence. However, limited information is available regarding use of OSCEs to assess competence in clinical/translational research. This study evaluated the feasibility and acceptability of an OSCE to assess progression toward competence of predoctoral students enrolled in three summer research programs (clinical/translational, global, and health disparities) at the University of Michigan. METHODS/STUDY POPULATION: Core program competencies were defined and five competencies which were shared among the programs were selected for inclusion in an end-program OSCE. All enrolled students were asked to participate. OSCE scenarios required students to identify equitable research practices, write clinical/translational research questions, apply appropriate study design principles, identify responsible conduct of research practices, and obtain informed consent for participation in a research study from a standardized patient instructor. Performance was rated on a 3-point descending scale: 3 - no errors, 2 - minor errors, 1 - critical errors. RESULTS/ANTICIPATED RESULTS: Twenty-three students (85%) completed the OSCE in 2 hours. Student performance varied within and across stations. In general, writing translational research questions appeared to be the most challenging task and performance appeared to be strongest in the areas of primary focus of the students' programs. This finding suggests that “in-program” learning was applied in responding to the tasks presented in the OSCE scenarios. DISCUSSION/SIGNIFICANCE OF IMPACT: The OSCE was well accepted by students and feasible to administer in terms of time, cost, and personnel.
MENTORING BEHAVIORS THAT MOST INFLUENCE PROTÉGÉ OUTCOMES: LESSONS FROM THE GENERAL WORKPLACE LITERATURE
Delgado PL1, Lichtenstein MJ1, McManus LM1, Stappenbeck SK1 1University of Texas Health Science Center at San Antonio, San Antonio, TX, USA
OBJECTIVES/SPECIFIC AIMS: To compare research on mentoring and protégé outcomes in the business workplace setting with that in the medical literature in order to identify mentoring behaviors that are most important to protégé outcomes METHODS/STUDY POPULATION: A literature review of the general workplace literature regarding protégé outcomes from formal mentoring. RESULTS/ANTICIPATED RESULTS: A large scientific literature on mentoring and protégé outcomes exists for the business workplace environment. More than 40 studies including over 4000 subjects and three meta-analyses of the effect of mentors on protégé outcomes were reviewed. Two types of studies predominate: those that compare outcomes across protégé's and non protégé's and those that correlate mentor functions with protégé outcomes such as advancement, salary, and job satisfaction. While mentee/trainee/protégé characteristics are the strongest predictor of outcomes, certain mentor behaviors stand out as having the largest effect size (a) helping protégés acquire new job-related skills (b) challenging them to learn more (coaching), (c) raising protégé awareness of unrecognized career options and how to pursue them, (d) providing sponsorship, exposure, and visibility (opening doors), (e) helping increase protégé's sense of “self efficacy” (awareness and belief that one is competent within an area), (f) serving as a trusted guide for thinking through career decisions and (g) assessing performance and providing feedback. DISCUSSION/SIGNIFICANCE OF IMPACT: Mentoring in the general workplace environment has a small to medium effect on protégé outcomes. The mentoring behaviors with greatest effects should also be measured in academic medicine.
RESEARCH TRAINING FOR MEDICAL STUDENTS: IMPACT ON RESEARCH SELF- EFFICACY
Curran MA1, Black M1, Kelly C1, Golshan S1, Jeste DV1 1University of California, San Diego, La Jolla, CA, USA
OBJECTIVES/SPECIFIC AIMS: The objective of this study was to evaluate the effects on research self-efficacy of three separate NIH-funded medical student summer research training programs. METHODS/STUDY POPULATION: Participants were medical students who had completed one year of medical school at the first assessment and who were enrolled in a summer research training program. To assess research self-efficacy a scale developed by Schwarzer et al which used core competencies and an adapted scoring system from general self-efficacy scales was utilized. RESULTS/ANTICIPATED RESULTS: Research self-efficacy increased significantly after completing the summer research training. There was no significant interaction between year of data collection and the three program types, or for the main effect of year or program for the overall self-efficacy at the baseline. There was no significant association of improvement in trainees' self-efficacy with gender, type of college attended, type of research, or program type. DISCUSSION/SIGNIFICANCE OF IMPACT: A hands-on, mentored research training experience combined with didactic learning resulted in a significant increase in research self-efficacy among medical students. Trainees' self-efficacy improved most on research methodology and communication, followed by regulatory and organizational aspects. The study has limitations. The sample was not randomly selected, but consisted of students interested in research experience. We lacked a control group who did not participate in a summer research training experience. There were no objective measures of improvement. Lastly, it will be years before we can determine whether the training had an impact on career choice. Nonetheless, the training did improve their research self-efficacy.
TRACKING RESEARCH EDUCATION OUTCOMES: SAN ANTONIO 11-YEAR EXPERIENCE WITH MASTER OF SCIENCE IN CLINICAL INVESTIGATION (MSCI)
Lichtenstein MJ1, Stappenbeck SK1, Tumlinson L1, Mayorga R1, Delgado PL1, McManus LM1 1UT Health Science Center at San Antonio, San Antonio, TX, USA
OBJECTIVES/SPECIFIC AIMS: Determine MSCI graduate outcomes (papers, grants, careers in science). METHODS/STUDY POPULATION: MSCI graduate cohort analysis. Tracked publications/grants through PubMed/eReporter/CVs. MSCI tracking rate: 96%. RESULTS/ANTICIPATED RESULTS: From 2002–12, 166 students were accepted in MSCI: 100 graduated (60.2%); 27 currently enrolled; 31 withdrew or dismissed (18.6%). Time to graduation similar for both sexes (women median time 2.3 years [95% CI 1.8, 2.8 years, n = 56], men 2.0 yrs [95% CI 1.8, 2.4 yrs, n = 44]). No difference in ethnic/racial group time to graduation for Hispanics (median time 2.4 yrs [95% CI 1.8, 2.8 years], n = 27), Whites (2.3 yrs [95% CI 2.0, 2.4], n = 45), Asians (1.8 yrs [95% CI 1.8, 2.8], n = 24), or Blacks (2.8 yrs [95% CI 1.8, 2.8, n = 6). 72 graduates were from School of Medicine. Others from Schools of Health Professions, Dentistry, Pharmacy, Military Training Programs, Veterans Affairs. Graduates authored 985 articles (31.6% 1st author, 931 peer-reviewed). 28 graduates (and teams) obtained 161 grants in roles as PI, Co-PI, or Co-I (Mean 5.8 [SD 4.3]). 66 remain in academic, fellowship training, or research positions; 10 are in other research positions; 10 are staff physicians in teaching programs; 4 are students; 13 are in private practice. DISCUSSION/SIGNIFICANCE OF IMPACT: There were no systematic differences in graduates by ethnic/racial group or sex relative to progress in the MSCI program. High retention rates in productive, diverse translational research and academic careers exist among MSCI graduates.
TRANSFORMING RESEARCH BARRIERS INTO PROCESS CHANGE THROUGH THE PROTOCOL DEVELOPMENT PROGRAM
Miller TJ2, Chaitt DG1, Kopka SL2, Albert SM2, Young KG2, Sevastita VS2, Salahuddin CF2, Pierson JF1 1NIAID/NIH, Bethesda, MD, USA; 2SAIC-Frederick, Inc., Frederick, MD, USA
OBJECTIVES/SPECIFIC AIMS: The Protocol Development Program (PDP) at the National Institute of Allergy and Infectious Diseases (NIAID) was formed in response to a survey conducted by the National Institutes of Health (NIH), which highlighted an apparent need for protocol development support given the increasing regulatory demands on investigators conducting clinical research. The PDP supports navigation of protocols through the development and review processes by assisting with logistics and writing. METHODS/STUDY POPULATION: The protocol development milestones, from concept to IRB approval, were mapped at the program's outset. Key Performance Indicators were identified such as stakeholder satisfaction, protocol volume, and number of repeat customers using PDP services. The PDP uses resources within the organization and collaborates with other departments to improve the efficiency of the development process, eliminate redundancy, and provide up-to-date information on guidance and policies. Since inception, the process flow was modified to enhance customer services and reflect current practice. RESULTS/ANTICIPATED RESULTS: The PDP assisted with the development of 50 studies ranging from natural history to phases 0 through IV clinical trials. Customer feedback has been positive, with all respondents indicating they would use the PDP again and would recommend the services to other investigators. DISCUSSION/SIGNIFICANCE OF IMPACT: The PDP has served as a model for other NIH divisions interested in initiating a similar program. Other research centers may benefit from examining their organizational structure and envisioning ways in which process change and resource enhancement can positively affect the overall protocol start-up pathway.
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- SCHOLAR/TRAINEE ABSTRACTS
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
- Research Professionals Abstracts Best Practices
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
CO-MORBIDITIES AMONG PATIENTS WITH CA-MRSA IN HEALTH CENTERS
Tobin JN1,2, Kost RG2, Jenks N1, Balachandra S1, Okpukpara O1, Khalida C1, Tsang A1, D'Orazio B1, Coffran C2, Pastagia M2, Evering TH2, de Lencastre H2, Tomasz A2, Pardos de la Gandara M2, Coller BS2 1Clinical Directors Network, New York, NY, USA; 2The Rockefeller University Center for Clinical and Translational Science, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: Community-Acquired Methicillin-resistant Staphylococcus aureus (CA-MRSA) is an emerging infectious disease. This project builds a research collaborative among 6 NYC-area Community Health Centers (CHCs), Clinical Directors Network, a practice-based research network, & Rockefeller University Center for Clinical and Translational Science. METHODS/STUDY POPULATION: Six CHCs near a MRSA Hospital Surveillance Network are collaborating to 1) enroll 129 patients with skin & soft tissue infections (SSTIs) using CDC criteria 2) collect demographic, history, physical exam, photos, quality of life data 3) develop methods for wound & nasal specimen collection/transport to clinical & research labs 4) conduct microbiologic culture/antibiotic sensitivity & whole genome analysis 5) integrate the full spectrum of translational research in studying CA-MRSA from the genetic determinants of antimicrobial resistance to treatment/clinical outcomes 6) explore patterns of transmission & differences in outcomes. RESULTS/ANTICIPATED RESULTS: To date, 117 wound [38% MRSA+, 16% Methicillin Sensitive Staph aureus (MSSA+)] & nasal (16% MRSA+) specimens were cultured & tested; in 13% of cases, both wound & nasal samples carried MRSA; all cultures were susceptible to Tetracycline, Trimethoprim/Sulfamethoxazole, Vancomycin, Linezolid & Gentamicin. Co-morbidities associated with MRSA+ wound cultures were abscess (OR = 2.63 95% CI 0.93,7.69) & asthma (OR = 2.70 95% CI 1.02,7.14). DISCUSSION/SIGNIFICANCE OF IMPACT: In patients presenting to CHCs with SSTIs, the association of MRSA+ wound cultures with asthma suggest that airway disease and/or treatment-related factors (eg, inhaled corticosteroids) may predispose to CA-MRSA infection.
RANDOMIZED ASSESSMENT OF ALCOHOL CONSUMPTION WITH COMPUTERIZED ALCOHOL SCREENING AND BRIEF INTERVENTION FOR ADULT EMERGENCY PATIENTS
Lotfipour S1, Mortazavi R1, Byun I1, Anderson C1, Hoonpongsimanont W1, Alix K1, Patel H1, Do H1, Moreno F1, Chakravarthy B1, Weiss J2 1Center for Trauma and Injury Prevention Research, Department of Emergency Medicine, UC Irvine School of Medicine, Irvine, CA, USA; 2Department of Health Science, College of Health and Human Development, California State University, Fullerton, Fullerton, CA, USA
OBJECTIVES/SPECIFIC AIMS: The goal of this prospective randomized study is to measure the change in self reported alcohol consumption at 3 and 6 months following ED administration of computerized alcohol screening (CASI) and brief negotiated intervention (BNI) with referral to treatment as compared to standard of care for at-risk drinkers. METHODS/STUDY POPULATION: This is a prospective randomized study in a tertiary care university hospital. CASI was administered to ED patients 18 years or older by researchers using a bilingual computerized tablet. Subjects were randomized based on a random number generator. The researchers were blinded. Alcohol use was assessed by online follow-up at 3 and 6 months. The data was analyzed using Stata, and compared by demographic groups by descriptive statistics and the chi-square test for independence . Risk was calculated using AUDIT scores. RESULTS/ANTICIPATED RESULTS: It is notable that all patients scored in the at-risk group, but on follow up, all but 3 had lowered their AUDIT score. 7 Subjects who were barely high risk became low risk, while 8 patients who scored in the high risk range became low risk. There was no significant difference in the AUDIT score change between the CASI and CASI with BNI groups. DISCUSSION/SIGNIFICANCE OF IMPACT: At 3 month follow-up nearly all of the 15 of the 18 subjects who followed up had decreased their AUDIT score. At this time there is no a difference between the two groups.
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- SCHOLAR/TRAINEE ABSTRACTS
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
- Research Professionals Abstracts Best Practices
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
A MULTICENTER SCREENING TRIAL OF THE SAFETY AND EFFICACY OF RASAGILINE IN PEOPLE WITH ALS
Wang Y1, Moore D2, Katz J2, Saperstein D3, Walk D4, Simpson E7, Genge A10, Bertorini T9, Fernandes JA6, Jackson C11, Elman L8, Swenson A5, Swerdlow R1, Dimachkie M1, McVey A1, Herbelin L1, Macchi Z1, Barohn R1 1University of Kansas Medical Center, Kansas City, KS, USA; 2California Pacific Medical Center, San Francisco, CA, USA; 3Phoenix Neurological Institute, Phoenix, AZ, USA; 4University of Minnesota, Minneapolis, MN, USA; 5University of Iowa Hospitals and Clinics, Iowa City, IA, USA; 6University of Nebraska, Lincoln, NE, USA; 7The Methodist Hospital System, Houston, TX, USA; 8University of Pennsylvania, Philadelphia, PA, USA; 9University of Tennessee, Knoxville, TN, USA; 10McGill University, Toronto, ON, Canada; 11University of Texas Health Science Center, San Antonio, TX, USA
OBJECTIVES/SPECIFIC AIMS: The specific aim of this study is to determine whether rasagiline is safe in the patient population & slows disease. The second aim is to determine if mitochondria are affected by rasagiline using biomarker assays before & after treatment. METHODS/STUDY POPULATION: This is an open label study in ALS patients treated with rasagiline 2 mg daily for 12 months. The primary outcome is the change of the slope of ALSFRS-R compared with controls from completed trials. The secondary outcome uses assays for lymphocyte mitochondrial potential & apoptosis. RESULTS/ANTICIPATED RESULTS: Six month data for 31 patients showed a reduced deterioration not significant versus controls (N = 31; 11.5% reduction; p = 0.44). Significant JC-1 ratio for mitochondrial hyperpolarization (JC-1 before = 0.46, after = 0.61; p = 0.05) with oxidative stress (Mitotracker before = 304.67, after = 1622.25; p = 0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: Six month data for the 31 patients indicate that 2 mg daily Rasagiline is tolerated in the ALS patient population & biomarkers could aid in evaluation of drug-target engagement. Funding due in part by an investigator initiated grant from TEVA pharmaceuticals (Yunxia Wang, MD – PI) & grant UL1 RR 033179 (now UL1 TR 000001) from the University of Kansas Medical Center CTSA.
IMPACT OF CHOLECALCIFEROL REPLETION ON ERYTHROPOIETIN REQUIREMENTS IN VITAMIN D-DEFICIENT HEMODIALYSIS PATIENTS: PILOT DATA FROM A RANDOMIZED CONTROLLED TRIAL
Mehrotra A1, Heeger P1 1Mount Sinai School of Medicine, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: Vitamin D deficiency is common in hemodialysis (HD) patients. Uncontrolled studies suggest correction of Vit D deficiency is associated with decreased EPO requirements. The purpose of this study was to determine the effect of Vit D repletion on EPO requirements in HD patients. METHODS/STUDY POPULATION: 79 Vit D-deficient (25OH-D level <25 ng/mL) hemodialysis patients were randomized to receive D3 (n = 51) or standard of care (no repletion, n = 28) in a 2:1 ratio. Patients randomized to D3 received 50,000 IU/wk to a goal 25OH-D of > 35 ng/mL. Changes in Vit D level, hemoglobin, and EPO requirements were assessed at 6 months. EPO (Darbepoetin) doses were adjusted by nursing staff as per dialysis unit protocol (target Hb 10–12 g/dL). RESULTS/ANTICIPATED RESULTS: Baseline characteristics were similar between groups, as were baseline Vit D levels (median 12.9 vs 13.0, p = 0.936), baseline Hb (mean Hb 11.8 g/dL vs 11.5 g/dL, p = 0.272), and baseline EPO requirements (median Darbepoetin dose 40 mcg/wk vs 50 mcg/wk, p = 0.696). Patients randomized to D3 had a rise in 25OH-D levels at 6 months (12.9 to 39.2 ng/mL, p < 0.001, n = 31), with a corresponding fall in EPO requirements (40 to 25 mcg/wk, p = 0.028) despite no change in Hb (11.8 to 11.2 g/dL, p = 0.170). No change in Vit D level, Hb, or EPO dose was observed in control patients at 6 months (n = 19). Patients randomized to D3 did not experience hypercalcemia. DISCUSSION/SIGNIFICANCE OF IMPACT: Our preliminary data from this ongoing randomized controlled trial suggest that treatment of Vit D deficient dialysis patients with D3 is safe, effective, and results in lower EPO requirements. As nutritional Vit D supplementation is cheap compared to EPO, the routine correction of Vit D deficiency in HD patients may result in significant cost savings.
PHASE I TRIAL OF SEQUENTIAL INTRAVESICAL MITOMYCIN C AND BACILLE CALMETTE-GUERIN INSTILLATION FOR NON-MUSCLE INVASIVE UROTHELIAL CARCINOMA OF THE BLADDER
Svatek RS1, Morales E1, Mueller E1,2, Curiel TJ1, Basler J1,2, Hugen C2 1UT Health Science Center San Antonio, San Antonio, TX, USA; 2South Texas Veterans Health Care System, San Antonio, TX, USA
OBJECTIVES/SPECIFIC AIMS: Preclinical evidence indicates that prior mitomycin C (MMC) administration potentiates the activity of bacillus Calmette-Guerin (BCG) by increasing the uptake of BCG by urothelial cells. The aim of this study was to determine the safety and tolerability of sequential intravesical treatment with MMC followed by BCG for patients with urothelial cell carcinoma of the bladder (UCB). METHODS/STUDY POPULATION: Patients with intermediate- or high-risk BCG naïve UCB were considered eligible for the trial. MMC was instilled into the bladder for 30 minutes. The MMC was then removed and a 10 minute washout with 60 cc of sterile water and manual irrigation was performed. Then BCG was then instilled into the bladder for a 2 hour dwell time. This was performed weekly for a total of 6 weeks of treatment. Two dose levels of BCG were tested (½ dose and full dose) and two dose levels of MMC were tested (10 mg and 20 mg) in separate patients. RESULTS/ANTICIPATED RESULTS: Nine patients were enrolled and eight patients completed the planned therapy. One patient was only able to tolerate 5 of 6 weekly treatments due to fatigue and worsening urinary frequency. No grade 3 or 4 toxicity was observed. At 10 mg MMC and 1/2 dose BCG, the most common reported symptom was urinary frequency (2 patients). Fatigue was experienced in 1 patient at this dose. At 20 mg MMC and full strength BCG, the most common symptom was urinary frequency (2 patients) and self-limited mild hematuria in 1 patient. DISCUSSION/SIGNIFICANCE OF IMPACT: Sequential combined treatment of bladder cancer with weekly MMC and BCG is safe and results in tolerable urinary frequency. The recommended dosage for phase II studies is 20 mg MMC and full strength BCG.
SAFETY OF TRANSCRANIAL DIRECT CURRENT STIMULATION IN PEDIATRIC HEMIPARESIS
Gillick BT1, Krach LE1,2, Feyma T2,1, Aldahondo N2, Menk J1 1University of Minnesota, Minneapolis, MN, USA; 2Gillette Children's Specialty Healthcare, St Paul, MN, USA
OBJECTIVES/SPECIFIC AIMS: Transcranial Direct Current Stimulation (tDCS) is a form of stimulation that has shown beneficial behavioral effects in adults with strokes, however, the use of tDCS has not been extensively studied in children. Our specific aims are to assess 1) tDCS safety and 2) potential change in paretic and nonparetic hand and cognitive function in children with congenital hemiparesis. METHODS/STUDY POPULATION: Twenty children will be randomly assigned to either a real or sham tDCS group. The intervention will consist of cathodal contralesional/anodal ipsilesional tDCS applied to the motor cortex at 0.7–1.0 mA for 10 minutes. Each subject will receive a battery of safety and functional assessments. RESULTS/ANTICIPATED RESULTS: This study is actively recruiting subjects. With conservative tDCS dosaging, it is anticipated that tDCS will not produce any major adverse events, including seizure activity. No projected change in paretic or nonparetic hand or cognitive function is anticipated. DISCUSSION/SIGNIFICANCE OF IMPACT: The use of tDCS alone or combined with behavioral intervention has been studied in adults with stroke and shown to have beneficial outcomes, yet very little research has been done on the safety of tDCS in children. Data obtained from this study will support further research examining the safety, efficacy and mechanisms of serial tDCS intervention combined with behavioral rehabilitation in pediatric hemiparesis. The use of noninvasive brain stimulation in conjunction with rehabilitation training intervention for a child with hemiparesis proposes a synergistic approach to improving hand function and enhancing quality of life.
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- SCHOLAR/TRAINEE ABSTRACTS
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
- Research Professionals Abstracts Best Practices
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
CAPACITY TO CONSENT TO RESEARCH IN EMERGENCY DEPARTMENT (ED) PATIENTS WITH ACUTE PAIN
Cowan E1,2, Klerman H1, Ma J1 1Albert Einstein College of Medicine, Bronx, NY, USA; 2Jacobi Medical Center, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: Controversy surrounds the capacity of patients in acute pain to consent to research. Instruments to assess capacity exist but have not been tested in ED patients or those in acute pain. We aimed to test the Macarthur Competence Assessment Tool for Clinical Research (MacCAT-CR) in ED patients with acute pain to determine if these patients meet the ethical imperative of informed consent, which requires both decisional capacity and understanding. METHODS/STUDY POPULATION: A convenience sample of English speaking ED patients ages 18 to 60 with triage pain scores of 1 or more were approached for study entry. Participants completed a demographic questionnaire and a MacCAT-CR interview. Interviews were independently scored by two investigators. We used a score of 15 or more on the understanding subscale to reflect adequate understanding. Correlation between MacCAT-CR scores and pain scores were examined using Spearman's rho. RESULTS/ANTICIPATED RESULTS: Fifty-three patients were approached of which 19 refused to participate. Mean age was 37 ± 12 years. Most (53%) were Hispanic or African American (29%). More women (71%) were enrolled because they were preferentially placed in private rooms where interviews were easier to perform. Triage pain scores were high (mean 7 ± 2). The median score was 20 out of 36 (IQR 16–23) on the MacCAT-CR understanding subscale, 5.5 out of 6 (IQR 4–6) on the appreciation subscale and 4 out of 6 (IQR 3–5) on the reasoning subscale. Most subjects (79%) demonstrated adequate understanding based on our a-priori cut point. There was no correlation between pain score and performance on any MacCAT-CR subscale. DISCUSSION/SIGNIFICANCE OF IMPACT: Despite being in moderate to severe pain most ER patients demonstrated capacity consent to research. Pain scores did not correlate with performance on any of the MacCAT-CR subscales.
EXPERIENCE WITH A NEW APPROACH TO RESEARCH ETHICS LEARNING
Yarborough M1 1University of California Davis, Sacramento, CA, USA
OBJECTIVES/SPECIFIC AIMS: Describe revisions to an existing RCR course Identify course learning objectives Explain select learning strategies List future additions to the curriculum METHODS/STUDY POPULATION: CTSA Scholars in Training RESULTS/ANTICIPATED RESULTS: A deeper level of engagement with research ethics by learners. An understanding that compliance with research regulations constitutes only a part of conducting research ethically. An appreciation of what constitutes trustworthy research and why conducting research in a trustworthy manner is important. DISCUSSION/SIGNIFICANCE OF IMPACT: Most research ethics learning consists of meeting NIH RCR mandates and these mandates contribute to a disproportionate emphasis on bad scientists and the behaviors they engage in. We are exploring a fundamentally different approach to organizing and delivering research ethics education. We are seeking ways to address knowledge, skills, behaviors and aspirations that all contribute to trustworthy, excellent science. A mix of learning approaches is being employed and efforts are underway to expand research ethics learning from the current course into a larger curriculum.
TRANSLATIONAL RESEARCH ETHICS EDUCATION NEEDS ASSESSMENT
McKinney E1 1University of Texas Medical Branch, Galveston, TX, USA
OBJECTIVES/SPECIFIC AIMS: Readers will: 1. Gain insight into the investigator's perspective on translational research and ethics. 2. Recognize the need to enhance existing ethics education programs to meet the evolving needs of translational researchers. 3. Learn how to begin to address researchers' concerns about ethics education for translational research. METHODS/STUDY POPULATION: Study Population: Twenty (20) leaders in the Institute for Translational Sciences at University of Texas Medical Branch Methods: Face-to-face semi-structured interviews, digitally recorded and transcribed Analysis: Qualitative methods. RESULTS/ANTICIPATED RESULTS: Leaders' knowledge of basic concepts of responsible conduct of research (RCR) was widespread. However, many leaders tend to equate fulfillment of regulatory requirements with ethical conduct of research and ethical treatment of volunteers or those who otherwise provide samples, data, and support for biomedical research. Broadening perspectives and finding productive ways to negotiate, collaborate, and share information, recognition, and benefits are among the challenges that must be addressed in translational research ethics education. Leaders note that they will have to do a better job of leading by example to fulfill ethical goals in translational research. DISCUSSION/SIGNIFICANCE OF IMPACT: With the change in focus of biomedical research from silo driven to multidisciplinary team-based research using vast biobanks, big data, far-reaching consortia, and “omics” of various stripes comes the need to consider how best to achieve respect for persons, beneficence, and justice for all who contribute to and benefit from the research. This study begins small by examining where we are in one CTSA institution, yet may act to spur others toward finding better options for preparing current and future researchers to conduct ethical translational research.
TRANSLATIONAL SCIENCE AND THE SELF-IDENTITY OF THE SCIENTIST
Kotarba JA1,2 1Institute for Translational Science, Galveston, TX, USA; 2Texas State University, San Marcos, TX, USA
OBJECTIVES/SPECIFIC AIMS: Translational science is changing the ways scientists accomplish their work, with greater emphasis on formal tasks such as novel methodologies and creative procedures for health care delivery. A key feature of the related cultural change is the way translational science impacts not only what the scientist does, but also how the scientist perceives him or herself. My aim is to examine the process of self-identity change among translational scientists. METHODS/STUDY POPULATION: I have to date conducted a series of individual, conversational-style interviews over the course of a year with 45 scientists. I organize my analysis in terms of the narratives or stories the scientists relate about themselves, their careers, and their work. RESULTS/ANTICIPATED RESULTS: The translational science movement creates a series of contrasts between the traditional and the evolving self-identity. Translational science now places expectations on scientists to become specific kinds of leaders with organizational and business skills as well as scientific skills. A common concern among them is the potential loss of key features of their self-identity, especially that of the intellectual. DISCUSSION/SIGNIFICANCE OF IMPACT: The scientists generally feel that they can become organizational/team leaders with acquired business acumen. Yet, they voice a bit of caution for depending totally on rationalistic strategies for essential processes such as creativity and innovation. Translational science leadership can perhaps enhance the accomplishment of these essential processes by acknowledging as well as nurturing the humanistic and intellectual sense of self important to many scientists.
Health Services Research
- Top of page
- SCHOLAR/TRAINEE ABSTRACTS
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
- Research Professionals Abstracts Best Practices
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
THE INFLUENCE OF TRADITIONAL HEALERS AND “HEAL THYSELF” RELIGIONS ON PATIENT LINKAGE TO HIV TESTING SERVICES IN ZAMBÉZIA PROVINCE, MOZAMBIQUE
Audet C1, Blevins M1, Rosenberg C2, Farnsworth S2, Salato J2, Moon TD1, Vermund SH1 1Vanderbilt University- University School of Medicine & Institute for Global Health, Nashville, TN, USA; 2Friends in Global Health, Maputo, Mozambique
OBJECTIVES/SPECIFIC AIMS: To identify and quantify potential sources of delay to HIV testing among symptomatic, newly-diagnosed HIV positive patients in southern Zambézia Province, Mozambique. METHODS/STUDY POPULATION: We conducted a cross-sectional study in southern Zambézia Province as to the health-seeking behavior of 530 symptomatic adults who recently tested HIV-positive. We conducted pairwise comparisons used chi-squared statistic and rank sum tests. Multivariable linear regression with log-transformed delay to testing, including district, age, gender, travel time to clinic, care from traditional healer, and care from a religious leader. RESULTS/ANTICIPATED RESULTS: We found that 60% of newly diagnosed HIV positive patients first sought care from traditional healer(s). Visiting healer(s) before seeking care from the clinic was associated with a 228% increase in time until testing. Those who saw >1 healer were delayed a median of 315 days (compared with 120 days for those who saw only one healer). DISCUSSION/SIGNIFICANCE OF IMPACT: Use of traditional healers is significantly associated with delays in HIV testing in rural Mozambique. When healers do refer, patients are very likely to go to clinic. Despite extensive training on identifying HIV symptoms in these districts, very few healers refer patients for ART. Our data suggest that education of healers is not sufficient– a new model to incentivize healer referral behavior is needed.
WHO IS COUNSELING ADOLESCENTS ABOUT INTRAUTERINE CONTRACEPTION?
Rubin SE1, McKee MD1 1Albert Einstein College of Medicine, Bronx, NY, USA
OBJECTIVES/SPECIFIC AIMS: Use of the intrauterine contraceptive device (IUD) decreases adolescent pregnancy rates, yet only 5% of contracepting U.S. adolescents have IUDs.The low IUD utilization is multi-factorial and includes primary care providers (PCPs) reluctance to offer IUDs to teens.Our objective was to identify factors associated with a PCPs' frequently counseling adolescents about IUDs. METHODS/STUDY POPULATION: An internet survey was sent to Pediatricians (Peds), Family Physicians (FP) and Obstetrician-Gynecologists (Gyn) associated with an academic medical center in the Bronx, NY. Respondents were asked about knowledge, practice and attitudes around IUDs and adolescents with a focus on PCPs' perception of teen pregnancy and sexually transmitted infection (STI) risk. We conducted univariate analyses by PCP type and logistic regression with our dependent variable “In a typical office visit conversation with a female adolescent about contraception, how frequently do you include information about IUDs”. We will conduct further multivariate analyses. RESULTS/ANTICIPATED RESULTS: Univariate analysis shows that Peds are least likely to: (1) learn about IUDs in training, (2) counsel adolescents about IUDs, and (3) feel competent with IUD counseling.There was no significant difference between the PCPs and their attitudes around IUDs, adolescents and STI risk.Preliminary logistic regression results show that PCPs who frequently counsel adolescents about IUDs (1) feel more competent with IUD counseling and management of side effects and (2) do not think IUD use, as compared to contraceptive pill use, decreases teens' condom use. DISCUSSION/SIGNIFICANCE OF IMPACT: Preliminary results suggest that interventions aimed at increasing PCP competency in IUD counseling and management may increase the proportion of PCPs who routinely counsel their adolescent patients about IUDs.
- Top of page
- SCHOLAR/TRAINEE ABSTRACTS
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
- Research Professionals Abstracts Best Practices
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
A CALL TO URGENT ACTION ON STANDARD ONTOLOGIES FOR TRANSLATIONAL SCIENCE
Hogan WR1, Brochhausen M1, Smith B2 1University of Arkansas for Medical Sciences, Little rock, AR, USA; 2University at Buffalo, Buffalo, NY, USA
OBJECTIVES/SPECIFIC AIMS: The Clinical and Translational Science Award (CTSA) Program has always emphasized data standards to promote sharing and comparison of data across the CTSA Consortium and beyond. Yet creation and adoption of such standards is still painfully slow. Urgent action remains necessary. METHODS/STUDY POPULATION: History shows the high value of standard terms, definitions, and symbols (i.e. ontology) to science. But the creation and adoption of such standards often takes decades, as we show with two examples: chemical elements and units of measure. Translational science requires a consistent set of standard ontologies spanning all scales, from molecule to organism to population. But clinical terminologies at the macro-scale—such as SNOMED and ICD—inhibit translational science. They are inconsistent with successful micro-scale ontologies such as the Gene Ontology, and they also cannot change rapidly with the advance of science. RESULTS/ANTICIPATED RESULTS: Translational science must settle on standards that evolve in a way that is closely tied to scientific advance. In the case of chemical symbols and SI Units adoption proceeded in three overlapping stages. First came widespread recognition and understanding of the problem. Second, influential stakeholders helped to develop, test, and select appropriate standards. Third, once scientifically useful standards emerged, the community enforced them via peer review. DISCUSSION/SIGNIFICANCE OF IMPACT: How can we accelerate progress on clinical ontologies through all three stages? The authors, all of whom are involved in the new CTS Ontology Affinity Group, sketch a plan for the Consortium to invest in and adopt standard clinical ontologies that are open and well disseminated, following the strategies of the Open Biomedical Ontologies Foundry, Neuroscience Information Framework, and eagle-i/VIVO/CTSAconnect communities.
DIMENSIONAL CLASSIFICATION OF SCHIZOPHRENIA (D.C.).THE NEUROCOGNITIVE ENDOPHENOTYPES (N.E.)
Estani P1 1Professor Dr.Leon S.Morra Medical Hospital., Córdoba, Argentina
OBJECTIVES/SPECIFIC AIMS: The actual classifications of mental disorders by multiple categories, such as the DSM IV TR, (APA, 1994), have stopped being effective, specially in the field of the new psychiatric genetics, neurobiological investigations of schizophrenia and mental disorders in general, because they do not fit in our actual neurobiological data in field. At the moment we are looking for simple, dimensional classifications (D.C.) cradles in the concept of endophenotype, as this concept was defined by Dr. Gottesman in its application to psychiatric research. (Gottesman,2003). METHODS/STUDY POPULATION: A total of 1200 participants (patients with a diagnosis of schizophrenia and normal controls) were the subjects. Neuropsychological tests were applied. We operationalized two neurocognitive endophenotypes (N.E.) : sensorial gating and cognitive performance. RESULTS/ANTICIPATED RESULTS: The dimensional classification permits the inclusion of the mechanism of neurocognitive endophenotypes . DISCUSSION/SIGNIFICANCE OF IMPACT: The dimensional classifications of mental disorders allow full inclusion of the concept of endophenotypes,defining the genotypes to benefit the understanding and treatment of schizophrenia. The full explanation of this categorization will be presented along with the results of this pilot sample.
EVALUATING IMPLEMENTATION OF EVIDENCE-BASED CHILDHOOD ASTHMA PRACTICES IN PRIMARY CARE
Markus A1, Gerstein MT1, Lyons V1 1George Washington University, Washington, DC, USA
OBJECTIVES/SPECIFIC AIMS: We offer a viable approach for the evaluation of implementation activities in primary care through our experience investigating the translation of evidence-based childhood asthma interventions in federally qualified health centers (FQHCs). METHODS/STUDY POPULATION: Three FQHCs (and three matched comparison FQHCs) were selected for participation in this study to reflect differing contextual and organizational features. FHQCs were engaged in participatory planning to identify a minimum set of evidence-based asthma counseling activities and data measures for implementation across study sites. Using a strengthened case study approach, we are gathering concurrent and prospective qualitative and quantitative data from multiple sources over multiple points in time to assess the process of implementation at each FQHC while simultaneously evaluating patient-level outcomes. Data will be integrated and plotted on a timeline so that results of clinical and cost effectiveness evaluations are linked directly to decision points at the FQHC and to the influence of the context. RESULTS/ANTICIPATED RESULTS: Preliminary findings show relatively seamless integration of consensus driven asthma interventions into varying real-world practices. DISCUSSION/SIGNIFICANCE OF IMPACT: The case study has been emphasized as a data collection method for exploratory research rather than as a research strategy for explanatory inquiries, but our experience suggests that a strengthened case study approach that uses a common protocol across study sites offers a rigorous and pragmatic design with significant advantages for implementation research. Triangulating process data with clinical and cost effectiveness measures will allow us to characterize how successful implementation occurs across varying contexts in order to better inform what practitioners and policymakers might replicate and disseminate to benefit the system at large.
TEAM SCIENCE IDENTIFIES NOVEL DRUGS FOR NEURODEVELOPMENTAL DISEASES
Bigio B1, Sonaky V1, Wittkowski KM1 1The Rockefeller University, New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: GWAS for complex diseases have had limited success. Analyzing SNPs individually requires several large studies to integrate the often divergent results. Methods based on assumptions (independence, linearity) motivated by computational convenience, rather than biological plausibility, often cause false positives. Teaming up statistics, genetics, and computer science provides more powerful approaches to identify genetic risk factors for common diseases. METHODS/STUDY POPULATION: In the presence of epistasis between SNPs, intragenic regions, or genes, statistical approaches based on the linear model (stepwise logistic regression) often have low sensitivity and generate an abundance of artifacts. Advances in distributed and parallel processing spurred development of novel non-parametric statistics. U-statistics for multivariate data (μGWAS) are not confounded by unrealistic assumptions, can now incorporate information about epistatic relationships (sequence of neighboring SNPs, recombination hotspots) to increase power, and use novel strategies to guard against artifacts. RESULTS/ANTICIPATED RESULTS: We present a reanalysis of autism data publicly available at dbGaP. Where previous analyses were inconclusive, μGWAS confirms the role of the Ras pathway and identifies a novel class of genes, suggesting repurposing of existing drugs. The computational environment is accessible for investigators at http://mustat.rockefeller.edu. DISCUSSION/SIGNIFICANCE OF IMPACT: Reducing artifacts caused by inappropriate models reduces the need for independent studies. For personalized diagnostics and treatment to live up to their full potential, cases and controls need to be closely matched to the patient involved. Adequately controlling for environmental and social confounders is much easier to achieve when only a few hundred subjects, rather than several thousands need to be matched.
WORKING COLLABORATIVELY; DEFINING PRACTICE PARAMETERS, WORKLOAD TOOL, STANDARD AND ABOVE STANDARD SERVICES
Hall K1, Anderson E1, Jordan KS1, Radziejowska J1 1Harvard Catalyst, Boston, MA, USA
OBJECTIVES/SPECIFIC AIMS: The Harvard Catalyst CTSC was established in May 2008 under a new NIH structure that brought together four existing General Clinical Research Centers under one granting institution. Harvard Catalyst provides a structure that encourages cross-institutional collaboration across five sites for research projects. The Harvard-affiliated administrative, nurse and nutrition directors determined that defining the respective staff time per study visit is essential for allocating resources consistently across sites. METHODS/STUDY POPULATION: Each of the three disciplines met to develop a standardized workload measure for each discipline, and then later defined a rationale and methodology for standard and above-standard services for each discipline. Collaboration took place between the groups to gather feedback, and ultimate vetting took place at the Harvard Catalyst Executive Committee, before the communication to the larger research community. RESULTS/ANTICIPATED RESULTS: We will share the specifics of the journey that each discipline took to develop their respective workload measure, as well as their initial, and revised standard and above standard definitions for cost recovery. At two of the five CRCs, the workload measurement tool has been implemented, with the remaining sites being implemented in 2013. DISCUSSION/SIGNIFICANCE OF IMPACT: The workload tool for each discipline allows us to compare visits across sites and the data can be used for budgetary and staffing purposes. In the future, all of the Harvard Catalyst CRCs will be utilizing the same methodology for defining Standard and Above Standard Services, and charging for Above Standard Services based on these principles. This methodology provides a transparent system for investigators and has facilitated cost sharing for services which are intense.
- Top of page
- SCHOLAR/TRAINEE ABSTRACTS
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
- Research Professionals Abstracts Best Practices
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
COMPARING INFORMAL CAREGIVERS OF CANCER AND NON-CANCER PATIENTS: THEIR BURDENS AND THEIR DESIRE TO TRAIN FOR FUTURE FORMAL CAREGIVING
Phillips SS1, Simon MA1, Nonzee NJ1 1Northwestern University, Chicago, IL, USA
OBJECTIVES/SPECIFIC AIMS: To better support current informal caregivers and help past caregivers transition back into the workforce, we studied the unique burdens of caregivers (CG) of both cancer (CA) and non-cancer (NC) patients. METHODS/STUDY POPULATION: We interviewed 30 CG (8 CA CG and 22 NC CG), gathering 99 closed-ended questions, 3 open-ended questions, and a 14 question caregiver burden assessment. RESULTS/ANTICIPATED RESULTS: First, more CA CG maintained full-time work after beginning caregiving than NC CG as evidenced by a 25% decrease in full-time work for CA CG compared with a 54% decrease for NC CG. Secondly, similar percentages of CA CG (50%) and NC CG (45%) said the cost of illness and medical care was “A great hardship” (from options: “A great hardship,” “A little hardship,” “A moderate hardship,” “No hardship”). However, a striking 25% of CA CG as opposed to 59% NC CG said they expected the cost of illness and medical care by the time the patient no longer needs care to be “A great hardship.” Thirdly, similar percentages of CA CG (57%, n = 7 due to one non-responder) and NC CG (55%) said they would like to get training to work outside the home caring for patients in other households. Lastly, we observed that CA CG experienced more extreme emotional and lifestyle burdens than NC CG as evidenced by the caregiver burden assessments. DISCUSSION/SIGNIFICANCE OF IMPACT: The emotional burden of CA CG is greater than NC CG while the expected financial burden is lower for CA CG compared with NC CG. Surprisingly, although the emotional burden of CA CG exceeds NC CG, both groups are equally interested in training to transform their informal caregiving skills into formal caregiving skills. Current caregiving support programs and post-caregiving training programs should strive to meet these demonstrated needs of both cancer caregivers and non-cancer caregivers.
COST AND OUTCOMES OF PUBLICALLY-INSURED KIDNEY TRANSPLANT RECIPIENTS BY BODY MASS INDEX
Ercole P1, Buchanan PM2, Lentine KL2, Brruoughs TE2, Schnitzler MA2, Modanlou KA3 1Goldfarb School of Nursing, Saint Louis, MO, USA; 2Saint Louis University, Saint Louis, MO, USA; 3University of Tennessee / Methodist Transplant Institute, Memphis, TN, USA
OBJECTIVES/SPECIFIC AIMS: Kidney transplantation recipients' recovery is exacerbated by non-ideal health prior to surgery. Given a growing prevalence of obese transplant candidates, research on the impact to posttransplant costs and outcomes of recipients along the full scale of body mass index (BMI) is essential. METHODS/STUDY POPULATION: This retrospective cohort study design included data for Medicare patients linked with records from the Organ Procurement and Transplantation Network and the United States Renal Data System to examine costs and health outcomes following kidney transplantation. Costs and outcomes were compared by recipient BMI. Posttransplant cost, defined as insurance charges, was the primary outcome of interest. Medical charges were summed during transplant hospitalization and each of three years following transplantation. Additional outcomes included delayed graft function (DGF), graft failure, patient survival, and length of transplant hospitalization. RESULTS/ANTICIPATED RESULTS: Accumulated costs were significantly enlarged for both extremely morbidly obese and underweight recipients. Extremely morbidly obese recipients also had inferior rates of graft and patient survival. Unadjusted rates of DGF were significantly increased for recipients with elevated BMI. DISCUSSION/SIGNIFICANCE OF IMPACT: Kidney transplant recipients with non-normal BMI should be of concern. Weight management programs aimed at waitlist candidates may reduce posttransplant costs and improve long-term outcomes. BMI may not be the most accurate assessment of body composition. The cross-sectional nature of this research prohibits determining causality of BMI and outcomes.
EXPLORATORY ANALYSIS OF PATIENT PERCEPTIONS OF WORSENING HEART FAILURE SYMPTOMS
Reeder KM1 1Goldfarb School of Nursing at Barnes-Jewish College, St. Louis, MO, USA
OBJECTIVES/SPECIFIC AIMS: The purpose of this project was to explore symptom perceptions, reporting patterns, and treatment-seeking behavior prior to hospitalization. METHODS/STUDY POPULATION: Retrospective interviews were conducted with 60 patients hospitalized for HF. RESULTS/ANTICIPATED RESULTS: Over 2/3 of patients reported having early warning signs before overt symptoms; however, only 15% of patients attributed warning signs to their heart. Changes in self-management routines for diet (18.3%) and medications (13.3%) were identified as probable triggers of symptoms. Most often, patients reported having typical HF symptoms, including shortness of breath (70%), fatigue (51.7%), and swelling (28.3%). Although over half of the patients were nearly constantly bothered by their symptoms and felt they needed to contact their health care provider, only 22 (37.3%) patients contacted their provider. Delay times between health provider contact and hospitalization ranged from 1 hour to more than 14 days. Prior to hospitalization, 11 patients were seen by their health care provider in the outpatient clinic. After patient contact, wait times for outpatient clinic appointments ranged from 10 minutes to 37 days. The time between being seen by a health care provider in the outpatient setting and hospital admission ranged from 20 minutes to 14 days. Decisions to either contact providers or go to the hospital were most frequently made by patients, themselves and least frequently made by health care providers. DISCUSSION/SIGNIFICANCE OF IMPACT: Research focused on personalized interventions amenable to symptom-management issues in HF is needed.
- Top of page
- SCHOLAR/TRAINEE ABSTRACTS
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
- Research Professionals Abstracts Best Practices
- Clinical Epidemiology
- Clinical Trial
- Health Services Research
- Outcomes Research
- Translational Basic-to-Clinical
A FRAMEWORK TO DEVELOP A MULTI-INSTITUTIONAL NETWORK COLLABORATION TO ADDRESS HEALTH DISPARITIES THROUGH DISTANCE EDUCATION
Ortiz CA1, Shaheen M5, Zheng J6, Tse A4, Adigun JO7, Torres M2, Segarra B1, De Jesus L1, Velez H3, Estape ES1, Soto LE1 1University of Puerto Rico, San Juan, USA; 2Universidad Central del Caribe, Bayamón, USA; 3Ponce School of Medicine & Health Sciences, Ponce, USA; 4University of Hawaii, Hawaii, HI, USA; 5Charles Drew School of Medicine, Los Angeles, CA, USA; 6Morehouse School of Medicine, Atlanta, GA, USA; 7Meharry Medical College, Nashville, TN, USA
OBJECTIVES/SPECIFIC AIMS: The Distance Learning Networking Group (DLNG) is a collaborative initiative organized by the University of Puerto Rico, Medical Sciences Campus, and clinical and translational leadership, co-sponsored by National Institute of Health (CRECD, HiREC and PRCTRC) programs. The goal of the initiative is to help in developing a multi-institutional online network for the dissemination and sharing of clinical and translational training and educational resources through various technological methods. METHODS/STUDY POPULATION: The purpose of the group is to help determine the feasibility of distance education as a mode of instruction to introduce the multi-disciplinary components of health disparities. The group includes representatives from seven (7) different universities in USA with the challenge to define strategies to strength the multicultural, and multi-institutional online collaboration. RESULTS/ANTICIPATED RESULTS: The DLNG explore and promote the use of online platforms and information technologies in offering and evaluating courses among institutions. As part of our analysis and results of this network a multi-institutional evaluation rubric for online courses have been developed. DISCUSSION/SIGNIFICANCE OF IMPACT: The DLNG represents an opportunity to share and disseminate knowledge to help reduce health disparities. The DLNG continues to work to increase the value of the research collaboration and dissemination through the use of Technology.
EQUIPPING NURSES FOR CLINICAL RESEARCH: IMPLEMENTATION OF A TRAINING PROGRAM TO CONDUCT RESEARCH ON “PREFERENCES TO RECEIVE INFORMATION ABOUT PALLIATIVE CARE FOR ADULT PATIENTS”
Pittroff GE1, Hendricks-Ferguson V2, Boehmer V3 1Goldfarb School of Nursing at Barnes Jewish College, St. Louis, MO, USA; 2St. Louis University, St. Louis, MO, USA; 3Missouri Baptist Medical Center, St. Louis, MO, USA
OBJECTIVES/SPECIFIC AIMS: To evaluate the usefulness of training methods utilized to equip nurses to conduct clinical research. METHODS/STUDY POPULATION: As part of our larger study, a structured training program was provided by senior nurse investigators to equip (N = 9), 2 palliative nurse specialists to recruit and consent, and 7 staff nurses to collect data for a descriptive pilot study with palliative care (PC) patients and/or the primary decision makers. Our training program encompassed didactic content, group-reflective sessions, and role-playing activities. This educational research evaluation included a pre-training survey, a post-training survey and a final binary evaluation of all training methodologies including narrative responses. Univariate summary statistics were completed on demographic variables. Non-parametric Wilcoxon signed-rank tests were used to compare Likert-type items across study period. Alpha was preset at 5% for all testing of significance. RESULTS/ANTICIPATED RESULTS: All responses to Likert items (except two) demonstrated a statistical increase in agreement. Responses to binary evaluation questions reported a unanimous agreement. Thus, indicating adequacy of training to prepare staff nurses to participate in research. DISCUSSION/SIGNIFICANCE OF IMPACT: Implications of this evaluation have relevance for primary investigator researchers, practicing nurses engaged in clinical research, and clinical research institutions. Evaluation of our completed training procedures provides evidence to develop a standardized training program to plan a future larger study to evaluate PC preferences of adult patients with a life-threatening illness.
EXAMINING THE PSYCHOMETRICS AND UTILITY OF THE RESEARCH COMPETENCY SELF-REPORT TOOL
Baker CN1, Bevans K2,3, Blum NJ2,3, Leff SS2,3 1Tulane University, New Orleans, LA, USA; 2The Children's Hospital of Philadelphia, Philadelphia, PA, USA; 3Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA
OBJECTIVES/SPECIFIC AIMS: Research training for healthcare professionals improves patient care and facilitates translational research efforts, but little is known about how to best evaluate such training programs. This study describes the development and psychometric evaluation of a brief self-report measure designed to address trainees' research-related competencies. METHODS/STUDY POPULATION: The authors used a Participatory Action Research approach to design a tool to evaluate the effectiveness of a MCHB-funded Leadership in Education in Neurodevelopmental Disabilities (LEND) interdisciplinary research training program. The Research Competency Self-Report (RCSR) is a 12-item measure that assesses research competencies using a 10-point rating scale. Participants were LEND trainees from 10 cohorts (2002–2012) representing 15 healthcare disciplines (N = 173, 96.5% with at least a master's, 90.2% female, 75.3% White) who completed the RCSR before and after a one-year research training program. RESULTS/ANTICIPATED RESULTS: A two-factor exploratory factor model that differentiated between traditional and non-traditional research competencies explained 71.45% of the variance in items. Internal (αt = .95; αnt = .69) and test-retest (rt = .96; rnt = .90) reliability were adequate and construct and criterion validity were supported by evidence of expected differences among disciplines and over time. DISCUSSION/SIGNIFICANCE OF IMPACT: This study describes the development and evaluation of a brief, psychometrically sound self-report measure of perceived research competency, which can be utilized to evaluate the research training of healthcare professionals with the ultimate goal of informing the translation of research to practice and improving patient outcomes.
HIREC'S ENDOWMENT ADVANCES AND SUSTAIN RESEARCH CAREER DEVELOPMENT
Huertas A1, Estape E1, Diaz C1, Soto L1 1University of Puerto Rico, San Juan, USA
OBJECTIVES/SPECIFIC AIMS: advance clinical/ translational research education and career development, keeping abreast of scientific changes,establishing effective collaborations, maximizing resources, and focusing in developing research capacities activities aimed at eliminating health disparities. METHODS/STUDY POPULATION: A 15 m NIH-NIMHD Endowment Hispanics-In-Research Capability:SoHP & SoM Partnership (HiREC)was awarded in 2008 as a permanent fund to expand/sustain high quality research training programs and infrastructure.The core program for accomplishing its goals is the postdoctoral Master inClinical/Translational Research. An operational infrastructure responsible of the annual action plan implemnetation. RESULTS/ANTICIPATED RESULTS: multiinstitutional course, Health Disparities:Translational Research Approach, distance learning implementation, scientific article in CTS.Curriculum revision of the MSc addressed NIH guidelines, created MSc courses.Technology infrastructure: complex data analysis, distance education.Visiting Professorship and Visiting Endowed Chair Programs: promote mentoring and new funding resources. Awards to study in MSc: scholars from UPR, PonceSoM&HS, UniversidadCentraldelCaribe, Bayamon increased scope of research in Puerto Rico. DISCUSSION/SIGNIFICANCE OF IMPACT: HiREC is a permanent funding resource to develop research infrastructure at UPR/MSC, enhance research career development of scholars and faculty, to facilitate the transfer of scientific advances at a faster pace and improve current health policy and practice Acknowledgement:Endowment “Hispanics-In-Research Capability:SoHP&SoM Partnership (HiREC)”, NIH-NIMHDGrant S21MD001830.The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
INNOVATION IN TRANSLATIONAL RESEARCH ETHICS EDUCATION
Farroni JS1, Klugman CM2 1UT MD Anderson Cancer Center, Houston, TX, USA; 2UT Health Science Center San Antonio, San Antonio, TX, USA
OBJECTIVES/SPECIFIC AIMS: Translational research increases the pace of discovery, development and demand for clinical impact. The compressed timeline reduces opportunities for reflection and increases the need for consideration of ethical issues. Translational research ethicists assist researchers to engage on difficult issues through education and dialog. The aim of this self-paced podcast series is to illuminate ethical issues that arise in the conduct of translational research through audio narrative in a manner that captures philosophical and practical complexity. METHODS/STUDY POPULATION: The podcast series is composed of episodes that that highlights a current ethical issue in translational research. Episodes are 10–15 minutes comprised of directed interviews with researchers, clinicians, research participants, caregivers, and the public. RESULTS/ANTICIPATED RESULTS: Each episode is a rich account of stakeholder perspectives on difficult ethical dilemmas encountered in the research context such as data management and peer review. The podcasts provide an amalgam of the theoretical/philosophical underpinnings of these issues alongside the practical experience and perceptions that manifest in the real world. Listeners are more engaged in the material than they would be during a case presentation, class or seminar. DISCUSSION/SIGNIFICANCE OF IMPACT: The podcast will offer a resource that compliments current research ethics training and education. It is a medium by which issues can be experienced in a way that deepens one's understanding of the complex and challenging issues facing translational researchers across institutions, disciplines, and professional stages.
PILOT PROJECT: IMPLEMENTATION OF THE COURSE IN HEALTH DISPARITIES A RESEARCH APPROACH
Soto de Laurido LE1, Estape ES1, Acosta- Perez E1, Mayor AM2,1 1MSC.UPR, San Juan, USA; 2Universidad Central del Caribe, Bayamon, USA
OBJECTIVES/SPECIFIC AIMS: Describe the Pilot Project to implement the Health Disparities: A Translational Research Approach Course. Discuss the different phases in the implementation process: instructional design, videos, lecturers from inside and outside Puerto Rico, course content, format, communication with scholars. Discuss the achievement of the course objectives by units. Evaluate the role of the course coordinators. METHODS/STUDY POPULATION: This course was designed with a multidisciplinary and interdisciplinary focus to address translational research in health disparities. RESULTS/ANTICIPATED RESULTS: We achieved an enrollment of nine (9) scholars. Two of the scholars were of two universities outside the University of Puerto Rico. The use of technology was not user friendly. The course did not plan any synchronous experience, but it was necessary to carry it out. DISCUSSION/SIGNIFICANCE OF IMPACT: It is the first time that the Post-Doctoral Master in Clinical and Translational Research of the Medical Sciences Campus implemented a course completely On-line. We will describe the Pilot Project developed to implement the Health Disparities: A Translational Research Approach Course from October to December 2012. The course defined health disparities taking into consideration the historical context, determinants and theoretical frameworks among other topics. At the same time we will compare the experience of the scholars living in Puerto Rico and the one from one School of Medicine from United States. Evaluation outcomes will be presented and the different rubrics used during the teaching process.
PROJECT DEVELOPMENT TEAMS: THE OUTCOMES AND IMPACTS OF TEAM SCIENCE FACILITATING AND ACCELERATING TEAM SCIENCE
Sajdyk T1, Sors T2, Shekhar A1, Denne S1 1Indiana Clinical and Translational Sciences Institute, Indianapolis, IN, USA; 2Purdue University, West Lafayette, IN, USA
OBJECTIVES/SPECIFIC AIMS: The complexity and interdisciplinary nature of translational research often requires input by basic and clinical researchers as well as other scientific expertise to develop a project. To facilitate this process, The Indiana Clinical and Translational Sciences Institute implemented a program to deal specifically with connecting investigators to all these individual resources through a team mentoring program for team science called the Project Development Teams (PDTs). The purpose of this study was to determine the impact of the PDT process for overcoming the barriers to translational research. METHODS/STUDY POPULATION: Using a REDCap database and survey, information was gathered from investigators for over 400 projects. Data was analyzed for multiple metrics including return on investment (ROI), time to funding for junior faculty, and satisfaction of the program. RESULTS/ANTICIPATED RESULTS: Analysis of the data indicate a strong ROI for projects at all translational stages (T1 – T3), accelerated time to external funding by junior faculty who received PDT assistance, and high satisfaction levels for investigators regardless of actual dollars received from the PDTs. DISCUSSION/SIGNIFICANCE OF IMPACT: The results of our data, when taken together and compared to the information in the current literature, indicate that investigators working with the PDTs are more successful than would be expected. The impact of the PDTs is multifold. First, they provide critical assistance to junior faculty which decreases the amount of time for funding; second, they help develop translational projects so that grants are more likely to succeed; and third, the interaction is a positive experience for all people involved, which makes investigators more likely to use the teams for assistance.
TEAM SCIENCE CULTURE: OUR EXPERIENCE IN A MULTIDISCIPLINARY TEAM
Carter MA1, Kotarba JA3, Farroni JS2 1UT Medical Branch, Galveston, TX, USA; 2UT MD Anderson Cancer Center, Houston, TX, USA; 3TX State University, San Marcos, TX, USA
OBJECTIVES/SPECIFIC AIMS: Translational research supports the development of novel methods and processes needed to determine health priorities, foster community research involvement and develop best practices that improve human health. Its focus is on increasing the pace of discovery, building infrastructure, enhancing training opportunities and accelerating innovation. This model of scientific inquiry incorporates new expectations regarding scientific teams and the forms of collaboration needed to accomplish these goals. An often neglected topic is precisely how researchers with different disciplinary backgrounds can learn to leverage their strengths and expertise to accomplish the work of translational science. We offer a team science framework which guides the development of translational research culture and seeks to illustrate how convergence of multiple disciplinary perspectives that makes the promises of translational science possible. METHODS/STUDY POPULATION: We present our own experiences in forming a Team Science Study Group. We discuss our values as a group, means of communication, style of inquiry and management, as well as feedback mechanisms to produce tangible deliverables. RESULTS/ANTICIPATED RESULTS: We demonstrate the expectations for multidisciplinary teams whereby each member embodies the shared values of: pre-competitive transparency, fair stewardship of resources, relationship-based partnership, shared decision making of team goals, honesty, commitment to success of the team and adherence to high ethical standards. DISCUSSION/SIGNIFICANCE OF IMPACT: The benefits of forging these teams include fostering perspectives that enhance intellectual investment in ideas, increased value of networking based upon common goals, openness, improved knowledge sharing, and increased trust through enhanced interpersonal relationships.
THE DUKE HUMAN HEART BIOREPOSITORY
Bowles D1, Schechter M1, Piacentino V1, Njoroge L1, Feger B1, Moseley A1, Milano C1 1Duke University, Durham, NC, USA
OBJECTIVES/SPECIFIC AIMS: Most interventions for cardiac disease and heart failure have been (and continue to be) developed in animal models. Human models of cardiac dysfunction and disease are needed to systematically study the biology of heart failure and test the efficacy of possible treatments as well as to develop better treatment strategies. Our long-term goal is to develop a multidisciplinary scientific infrastructure that focuses on human heart tissue to enable the discovery, testing, and translation of drug, gene, cellular and organ transplant therapeutic modalities for heart injury, disease, and failure. METHODS/STUDY POPULATION: To reach this goal, we have created a readily accessible, well curated, clinically relevant human heart tissue repository. We are building the infrastructure to acquire and understand the data from these human tissues. We provide an example of how our group has utilized this resource to futher our research program. In our example, we compared the proteomic and phosphoproteomic profiles of heart tissue from patients diagnosed with either ischemic or non-ischemic heart failure to non-failing controls using mass spectrometry to better understand the molecular changes underlying heart failure. RESULTS/ANTICIPATED RESULTS: This study identified quantifiable differences in the cardiac proteome and phosphoproteome specific to heart failure etiology and further characterized the cellular pathways central to disease development. Continued identification of novel pathways and molecular targets through proteomic analysis of human heart tissue will help improve prognostication and therapeutics for HF. DISCUSSION/SIGNIFICANCE OF IMPACT: Required are the means to disseminate the tissue and acquired information to scientific and medical experts for further development and testing of treatment options for heart disease, predicting heart failure, and improving transplant outcomes.
THE MURDOCK STUDY: ENROLLING A DIVERSE POPULATION INTO A COMMUNITY REGISTRY & BIOREPOSITORY FOR PATIENT POPULATION CHARACTERIZATION
Nunes PB1, Cornish M1, Bouk L1, Smerek M1, Plez M1, Dunham A1, Wixted D1, Dolor R1, Christian V1, Califf R1, Newby L1 1DTMI-MURDOCK Study, Kannapolis, NC, USA
OBJECTIVES/SPECIFIC AIMS: (1) Articulate MURDOCK Study goals for building a large-scale epidemiological resource; (2) describe recruitment strategies for engaging a diverse community. METHODS/STUDY POPULATION: Persons 18 years and older, residing in selected zip codes in Cabarrus County region of NC are recruited by open enrollment. Participants complete a questionnaire about demographics, health-related quality of life, activity/lifestyle, and diet/nutrition. Vital signs, waist circumference and samples of blood and urine are banked for future research. Participants consent to yearly follow up, geospatial mapping and access to their medical/electronic health records. RESULTS/ANTICIPATED RESULTS: The MURDOCK Study has established strong, on-the-ground operations in select zip codes since 2009. Of the 9,000 participants already enrolled, 13% are African American and 9% Hispanic, a racial composition similar to Cabarrus County, NC. Engagement activities span community organizations, churches, schools, and businesses to reach a diverse population. A pilot representative sampling of households in two zip codes begins in January 2013 to evaluate operations before implementing a full-scale recruitment effort that will enroll 15,000 adult residents. DISCUSSION/SIGNIFICANCE OF IMPACT: The MURDOCK Study Community Registry and Biorepository is a valuable research engine and vehicle for the conduct of translational medicine. This population-based registry of health information, paired with banked biological specimens and access to EHRs, will serve as a viable foundation for research efforts across academic, private, governmental and other collaborators by providing a diverse and representative sample.
TRAINING THE NEXT GENERATION OF TRANSLATIONAL SCIENTISTS: THE UPENN EXPERIENCE
Meagher EA1, Bastian RA1 1University of Pennsylvania, Philadelphia, PA, USA
OBJECTIVES/SPECIFIC AIMS: The Master of Science in Translational Research (MTR) at the University of Pennsylvania's Perelman School of Medicine began in 2004 with a mission to produce a new cadre of translational scientists who are competitive in seeking research support and knowledgeable about the complex issues associated with conducting sound translational research. The program regularly collects outcome data to determine if the training program is achieving its mission of preparing these trainees for careers as independent investigators in academia. METHODS/STUDY POPULATION: Metrics used to evaluate the success of the MTR program include student and graduate career progression measured by number of grant awards, progression of academic appointment, and journal publications. The program is also evaluated qualitatively through student satisfaction. Data was collected from curriculum vitaes, NIH RePORTER, PubMed, academic enrollment records, and program evaluations. RESULTS/ANTICIPATED RESULTS: The overall MTR graduation rate is 78%. The rate in 2004–2007 was 65% and increased to 90% in 2008–2010. MTR graduates have been awarded 16 training grants (12 K23, 4 K08), 3 R01s, and 18 other federal grants. Additionally graduates were awarded 33 foundation and 15 industry grants. The majority of graduates remain in academia with 75% currently in academic faculty positions and 19% in trainee positions. Eighty MTR students have authored 528 publications. Qualitative measurements show 81% of students rated the overall program as very good or excellent, 100% of graduates rated the program helpful or very helpful in their career, and 85% rated the program helpful or very helpful in obtaining funding. DISCUSSION/SIGNIFICANCE OF IMPACT: The MTR program is meeting its primary objective to train translational scientists who are competitive in seeking research support and prepared for career paths to become independent investigators in academia.
UPR REVISED MASTER OF SCIENCE IN CLINICAL AND TRANSLATIONAL RESEARCH
Segarra B1, Estapé ES1, Huertas A1, Diaz C1 1University of Puerto Rico, San Juan, USA
OBJECTIVES/SPECIFIC AIMS: The post-doctoral Master of Science in Clinical Research of the University of Puerto Rico has been funded by NIH since 2003. Recognizing the importance of keeping this program abreast with innovation and the trends in translational research, a curricular revision process took place in 2009. The curricular revision was based on NIH guidelines that define knowledge, skills, and attributes that a master's-level scholar should attain while training to become clinical and translational researcher. METHODS/STUDY POPULATION: The MSc Faculty reviewed their courses aligning the course objectives with recommended NIH 103 competencies, and ascertained the level in which they were addressed (Generalist, Specialist, and Advanced). Gaps, strengths, and content overlapping were analyzed. RESULTS/ANTICIPATED RESULTS: The curricular revision led to a written academic proposal for the implementation of the post-doctoral Master of Science in Clinical and Translational Research Program. Three new courses were developed to fulfill the gaps: Scientific Communication in Clinical and Translational Research, Introduction to Biomedical Informatics, and Health Disparities: Translational Research Approach. Other minor modifications to existing courses also took place. DISCUSSION/SIGNIFICANCE OF IMPACT: The curricular revision allowed the faculty and MSc leadership to critically evaluate the courses' content, sequence, and its relationship with the new trends in translational research. Although most of the competencies recommended by NIH were reflected in the Program, the gaps were addressed by the development of new courses and other minor changes. The revised curriculum is characterized by its innovation in response to the current research education paradigm in clinical and translational research. Partially supported by the National Institute on Minority Health And Health Disparities of the NIH: R25MD007607 and HiREC S21MD00183.
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AMELIORATION OF EXPERIMENTAL DIABETIC NEPHROPATHY BY CURCUMIN IS ASSOCIATED WITH FUNCTIONAL AND HISTOLOGICAL IMPROVEMENT OF THE KIDNEY
Warraich I1, Park S1, Prabhakar S1 1Texas Tech University Health Sciences Center, Lubbock, TX, USA
OBJECTIVES/SPECIFIC AIMS: Diabetic nephropathy (DN) is the leading cause of end stage renal failure but currently there is no effective therapy for DN. Since curcumin (diferuloylmethane) had been shown to have antioxidant and anti-inflammatory effects especially in cancer, we investigated it's effects on experimental DN in a rat model. METHODS/STUDY POPULATION: ZSF rats- murine models of DN characterized in our laboratory, were used to test our aims and were maintained from 8th to 26th week and given either curcumin at 1 mg/ml in drinking water (test group) or plain water (control group). Urine and blood samples were obtained at the start and end of the study to examine the creatinine clearence (Ccr) and proteinuria. At 26 weeks, rats were sacrificed and kidneys harvested to examine the histopathology. RESULTS/ANTICIPATED RESULTS: At 8th week, all rats had normal renal functional parameters but by 26th week, control rats exhibited severe proteinuria (1458 ± 168 mg/G creat) and decline in Ccr (3.1 ± 0.7 L/Kg/BW/d).Curcumin treated rats had lesser proteinuria (862 ± 96 mg/G creat) and better preserved Ccr (4.3 ± 0.9 L/Kg/BW/d). Renal histology in control rats showed glomerular sclerosis (nodular and diffuse) with tubular atrophy, interstitial infiltration and severe arteriolar thickening. Curcumin treatment ameliorated glomerular and interstitial changes significantly while not affecting the vascular thickening. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results show that curcumin improved renal function in experimental DN and this was accompanied by significant amelioration of renal pathological changes of DN. We conclude that based on our experimental data, curcumin may have potential therapeutic benefits in human DN.
DEVELOPMENT OF HIGHLY SENSITIVE NANO-IMMUNOASSAYS TO DEFINE ABERRANT SIGNALING AND DRUG RESPONSE IN PEDIATRIC ACUTE MYELOID LEUKEMIA
Sabnis HS1, Bradley H1, Cooper T1, Bunting KD1 1Emory University - Aflac Cancer and Blood Disorders Center, Atlanta, GA, USA
OBJECTIVES/SPECIFIC AIMS: Disease relapse in acute myeloid leukemia (AML) is a major cause of mortality. Leukemic cell survival results from up- regulation of survival pathways such as JAK-STAT, Ras/Raf/ERK and/or the PI3K-Akt-mTOR pathway. Improvement in AML therapy relies on understanding the dysregulation of these key signaling nodes. Aim 1: To develop a sensitive nano-immunoassay to obtain a protein signaling profile in acute myeloid leukemia. Aim 2: To utilize the NanoPro 1000 to demonstrate effective inhibition of drug targets in mTOR and PI3K pathways. METHODS/STUDY POPULATION: The NanoPro 1000 system is an automated capillary-based immunoassay platform that enables rapid and quantitative phospho-protein analysis. It can compare multi-phosphorylation sites on the same protein and enables innovative analysis of potential new biomarkers and targets. We have examined several AML cell lines and have been able to profile key proteins involved in mTOR, ERK and PI3K cell survival pathways in AML. RESULTS/ANTICIPATED RESULTS: We will use the NanoPro to obtain real time 'activation signatures' in patient samples with an antibody panel comprising JAK-STAT, ERK and PI3K-Akt-mTOR targets. These signatures can be compared pre- and post therapy and at diagnosis and relapse. This will provide insight into the evolution of specific leukemia cell populations. We will treat primary samples with mTOR, ERK and PI3K inihbitors and use the NanoPro 1000 to demonstrate effective inhibition of drug targets. DISCUSSION/SIGNIFICANCE OF IMPACT: This platform has the potential to interrogate the biologic efficacy of new drugs with defined targets and our long term aim is to utilize this platform in de-novo AML to be able to identify poor responders who might benefit from early introduction of targeted therapy.
DISCOVERING LINEAGE-SPECIFIC BIOMARKERS FOR METASTATIC STEM CELLS IN LUNG ADENOCARCINOMA
Nguyen D1 1Yale School of Medicine, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: Cancer metastasis is a dynamic process fueled by heterogeneous malignant cell populations with numerous molecular alterations. These factors all contribute to variable clinical courses, likely requiring disease-specific therapeutic approaches. Lung adenocarcinoma (ADC) is a particularly aggressive subtype of lung cancer that will frequently metastasize despite treatment with curative intent. This implies the presence of therapeutically resistant malignant cells with a high degree of metastatic competence within primary lung ADCs. The cellular and molecular identity of this cell type(s) remains unknown. METHODS/STUDY POPULATION: To identify disease-specific biomarkers of early stage metastastic lung ADCs, we implemented a systematic computational analysis of gene expression patterns from normal pulmonary epithelial cell lineages, activated embryonic stem cells, annotated cohorts of patient lung tumors, and an experimental model of lung cancer metastasis that we previously developed. Through this integrated approach, we identified a gene module that specifically marks a subgroup of poorly differentiated metastatic stage I ADCs. RESULTS/ANTICIPATED RESULTS: These putative biomarkers include several known cell surface antigens whose expression correlates with epithelial regeneration and the metastatic potential of experimentally derived malignant cell populations. Thus these may endow tumor cells with self-renewing properties within unique tissue niches. DISCUSSION/SIGNIFICANCE OF IMPACT: Based on these results, we have characterized the cell surface expression pattern and functions of putative biomarkers for the identification, isolation, and characterization of metastatic stem cells during various stages of lung ADC progression and therapeutic response.
EFFECT OF ENDOTHELIAL CELL FUNCTION ON INTERDIALYTIC BLOOD PRESSURE IN HEMODIALYSIS PATIENTS WITH INTRADIALYTIC HYPERTENSION
Van Buren PN1, Toto R1, Inrig J2 1University of Texas Southwestern Medical Center at Dallas, Dallas, TX, USA; 2Duke University, Durham, NC, USA
OBJECTIVES/SPECIFIC AIMS: Intradialytic hypertension is an increase in blood pressure (BP) during hemodialysis (HD) associated with high mortality in HD patients. These patients have higher BP between HD treatments and worse endothelial cell function than HD controls with BP decreases during HD. We hypothesized endothelial cell function was associated with interdialytic BP patterns in intradialytic hypertension patients. METHODS/STUDY POPULATION: In a case control study of intradialytic hypertension patients and HD controls, we measured endothelin-1 (ET-1) and asymetric dimethylarginine (ADMA) before and after HD; interdialytic ambulatory BP; and flow-mediated vasodilation (FMD) on a non-HD day. Using mixed linear models, we determined associations of FMD and intradialytic changes in ET-1 and ADMA with ambulatory BP slopes. RESULTS/ANTICIPATED RESULTS: Of 50 subjects (25 per group), the mean age was 54 yrs, 80% were male, 38% African American, and 86% diabetic. Ambulatory BP was higher in patients with intradialytic hypertension than controls (155 vs. 145 mmHg, p = 0.005). For the first 24 hrs after HD, BP decreased in patients with intradialytic hypertension (–0.3 mmHg/hr, p = 0.1). In these patients higher FMD and higher ADMA removal during HD were associated with a steeper BP slope (β = –0.9 and –0.03, p = 0.07 and 0.003). In controls, BP increased throughout the interdialytic period (+0.5 mmHg/hr, p = 0.002). FMD and changes in ET-1 or ADMA had no effect on BP slope (β = 0.001, 0.002, –0.05; p = 0.9, 0.9, 0.8). DISCUSSION/SIGNIFICANCE OF IMPACT: Both acute and chronic endothelial cell dysfunction is associated with interdialytic BP patterns in intradialytic hypertension patients. Treatments that modify endothelial cell function may result in better overall management of hypertension in these patients.
FEASIBILITY OF QUANTIFYING INTRAMYOCARDIAL BLOOD VOLUME USING SPECT/CT
Liu C1, Chan C1, Stacy M1, Miller K1, Bregasi A1, Dione D1, Zhuang Z1, Sinusas A1 1Yale University, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: SPECT/CT quantification of intramyocardial blood volume (IMBV) using 99mTc-labeled red blood cell (RBC) can provide information on myocardial microcirculatory function. However, the RBC tracer concentration within the blood pool is much higher than the adjacent myocardium, causing substantial contamination to the myocardium because of partial volume effect (PVE), scatter, and respiratory/cardiac motion, resulting in substantial overestimation of IMBV. In this study, we developed methods to quantify IMBV and evaluated the IMBV responses to various vasodilators. METHODS/STUDY POPULATION: Respiratory motion was corrected by extracting listmode data from end-expiration phases, which were further rebinned into dual gated data to correct for cardiac motion. CT angiography was used to define the myocardium to facilitate region-based voxel-wise partial volume correction (PVC). To reduce noise while preserving quantification, an anatomical-based non-local means filter (ANLM) was applied. Triple energy window (TEW) method was used to correct for scatter. We studied the IMBV change as a response to various vasodilators in a dog study to investigate its sensitivity to the change of microcirculation function. All data were acquired using a dedicated cardiac SPECT/CT system (GE Discovery 570c). RESULTS/ANTICIPATED RESULTS: Corrections of PVC, respiratory and cardiac motion, and scatter effectively reduced the activity contamination to myocardium. PVC with ANML filter reduced image noise while preserving accurate quantification. In animal study, the IMBV increased by 22.5%, 9.7%, and 15.4% with adenosine, nitroglycerin, and dobutamine, respectively. DISCUSSION/SIGNIFICANCE OF IMPACT: With quantitative correction methods, it is feasible to accurately quantify IMBV using 99mTc-RBC SPECT/CT. The relative change of IMBV is sensitive to the intervention of microcirculation function using stressors.
IMMUNE RESPONSE ALTERATION BY THE ARGININE METABOLIC PATHWAY: IMPLICATION FOR THE ONSET OF FATIGUE INDUCED BY LOCALIZED RADIATION THERAPY
Wang X1, Hsiao C1, Reddy S1, Saligan L1 1NINR/NIH, Behtesda, MD, USA
OBJECTIVES/SPECIFIC AIMS: Fatigue is a common side effect of radiation therapy (RT) for cancer. The molecular mechanisms underlying radiation-induced fatigue remain unclear. This study investigated the differential expression of genes and pathways during the development or worsening of fatigue while receiving localized RT. METHODS/STUDY POPULATION: Thirty men with prostate cancer scheduled to receive external beam radiation therapy (EBRT) were recruited. Fatigue scores measured by 7-item PROMIS-fatigue scale and blood were collected prior to treatment (baseline) and at midpoint (day 21) of EBRT. RNA was used to conduct an unbiased whole genome microarray analysis using HG-U133+2.0 array. Differences in gene expression between timepoints were analyzed using ANOVA followed by post hoc and corrected by a filtering criteria of a > 2.0 or < -2.0 fold and p valus corrected for false discovery rate using using Partek® Genomic Suite. RESULTS/ANTICIPATED RESULTS: Fatigue scores increased significantly from baseline (44.9 + 5.8) to midpoint (49.1 + 5.3, p = 0.01) of EBRT. ARG1 (encoding arginase type 1) was significantly upregulated from baseline to midpoint of EBRT (fold change = 2.41, p < .05); whereas the genes associated with innate immune functional pathway (CD28, CD27, CD3D, CD8A and HLA-DOB) were significantly downregulated over 2-fold between the study timepoints. DISCUSSION/SIGNIFICANCE OF IMPACT: Increased expression of ARG1 results in arginine deficiency which leads to immune depression by impairment of lymphocyte proliferation and formation of the T-cell receptor. Our findings indicate that arginine deficiency-associated immunosuppresion may play a role in the development or worsening of fatigue during EBRT. These findings provide evidence at the transcriptional level that the use of arginase inhibitor or arginine supplementation may reduce the fatigue symptoms of this population.
INFLAMMATORY CYTOKINES CONTRIBUTING TO ALTERED INSULIN SIGNALING AND NEPHROPATHY ARE MEDIATED BY OXIDATIVE STRESS IN EXPERIMENTAL TYPE II DIABETES
Prabhakar S1, Sohal D1, Park S1 1Texas Tech University Health Sciences Center, Lubbock, TX, USA
OBJECTIVES/SPECIFIC AIMS: Since the inflammatory basis for diabetic nephropathy (DN) is not well understood, we hypothesized that oxidative stress (OS), an event in the pathogenesis of DN stimulates inflammation and contributes not only nephropathy but altered insulin signaling in DN. METHODS/STUDY POPULATION: Obese ZSF rats, a model of DN characterized in the author's laboratory, were used to test our hypothesis. Lean ZSF (non-diabetic hypertensive) rats were used as controls. We examined the effects of curcumin (diferuloylmethane), an antioxidant and losartan, a blocker of angiotensin and OS. Rats were maintained from 8th-26th week on high calorie diet and given curcumin at 2 mg/ml or losartan at 25 ml/L in water or just plain water. Rats were monitored for weekly wt. and urine output. Renal functional parameters urine albumin excretion (UAE) and creatinine clearance (Ccr) were measured at the start and end of the study. At 26th week rats were sacrificed to harvest kidneys and to examine the expression of NFkB, TNF-α, IL-6 (inflammatory markers), 8 isoprostane and 8 hydroxydeoxyguanosine (oxidative markers). We also examined Hb A1c, IRS-1 phosphorylation (pIRS-1) and adiponectin receptor (AdipoR) expression in the muscle as of measures of insulin signaling and sensitivity. RESULTS/ANTICIPATED RESULTS: Diabetic rats at 26th week had features of DN along with high levels of OS markers and also increased expression of NFkB, TNF-α, IL-6 in the kidney. Curcumin and losartan improved renal function and inhibited inflammatory markers by reducing OS. This was also associated with better insulin signaling shown by lower HbA1c, increased pIRS-1 and AdipoR expression. DISCUSSION/SIGNIFICANCE OF IMPACT: Inflammation in DN may be initiated by OS as antioxidants improve insulin signaling and DN by ameliorating oxidative stress and inhibiting inflammation.
MODELING NEUROMUSCULAR DISORDERS VIA INDUCED PLURIPOTENT STEM CELLS
Huttner A1, Bao Y1 1Yale University School of Medicine, New Haven, CT, USA
OBJECTIVES/SPECIFIC AIMS: The generation of human induced pluripotent stem cells (iPSCs) opened new avenues for human disease modeling. IPSCs allow the generation of large numbers of genetically modifiable cells specific to the underlying human genetic background. We focused on Walker-Warburg-Syndrome (WWS), a severe form of congenital muscular dystrophy, also associated with lissencephaly. Several genes have been implicated in the etiology of this syndrome, however, its pathogenesis is still poorly understood. In addition, none of the animal models appears to faithfully reflect the human condition. Patient derived IPSCs allow the targeted differentiation of cells into tissue specific phenotypes of brain and muscle, and thus provide an assay for the recapitulation of disease specific pathophysiology. METHODS/STUDY POPULATION: IPSC lines were derived from skin biopsy specimens of patients with WWS and normal age matched controls. Reprogramming of human fibroblasts was achieved with a lentiviral construct expressing OCT4, SOX2, KLF4 and c-Myc/mCherry. The cells were grown in-vitro and differentiated into neurons and skeletal muscle. The in-vivo potential of these progenitor cells was tested via in-utero transplantation, demonstrating the cells' ability to follow microenvironmental cues, migrate and differentiate appropriately. RESULTS/ANTICIPATED RESULTS: Directed differentiation of iPSCs into neuronal and myogenic precursors was demonstrated in vitro with antibodies for CNS phenotypes, like GFAP, TUJ1, Tbr1/2 as well as muscle phenotypes. The engraftment and integration of individual cells in-vivo showed the ability of iPSCs to differentiate appropriately and show a disease specific difference to their normal counterpart. DISCUSSION/SIGNIFICANCE OF IMPACT: This model allows the phenotypic recapitulation of complex neurogenetic traits, and provides insights into the pathophysiology of human forms of WWS.
RECURRENT FURUNCULOSIS IN A COMMUNITY ACQUIRED S. AUREUS INFECTION CAUSED BY A STRAIN BELONGING TO THE USA300 CLONE OF MRSA FUNDING: NIH-NCATS GRANT#8UL1-TR000043
Pardos M2, Balachandra S1, Urban T1, Salvato S1, Khalida C3, Kost R2, Tobin JN3,2, Tomasz A2 1Urban Health Plan, Bronx, NY, USA; 2The Rockefeller University, New York, NY, USA; 3Clinical Directors Network (CDN), New York, NY, USA
OBJECTIVES/SPECIFIC AIMS: NA METHODS/STUDY POPULATION: As part of a multicenter study on community acquired methicillin resistant Staphylococcus aureus (CA-MRSA) in the New York metropolitan area, we enrolled a previously healthy 24-year old female with recurrent skin and soft tissue infections (SSTI). Folliculitis and furuncles were first observed on her buttocks and thighs, evolving to cellulitis requiring ER admission and IV Vancomycin. One month later, lesions reappeared on the abdomen and persisted for one month. Three months after the first episode, folliculitis was observed on the abdomen and an abscess on the right elbow. Wound samples were taken in the first 3 episodes from the lower body and nares, oropharynx, axillae, groin, cubital and patellar folds were also sampled. DNA probes for the mecA determinant identified MRSA in all wound samples and in the groin, and one month later in the groin and the right patellar fold. No MRSA nasal carriage was detected at any time. RESULTS/ANTICIPATED RESULTS: The MRSA strain recovered from all locations belonged to spa type t008 with a PFGE profile characteristic of MRSA clone USA300, which also carried the virulence gene pvl and ACME type I. The strain was resistant to beta-lactams, erythromycin and levofloxacin, and susceptible to clindamycin, gentamycin, linezolid, tetracycline, co-trimoxazole and vancomycin. The patient was treated with co-trimoxazole and vancomycin. The latest episode was treated with doxycycline. During the first two episodes of infection the patient reported that a sister who shared the household had a similar SSTI and was treated in another facility, indicating potential household transmission or common exposure. DISCUSSION/SIGNIFICANCE OF IMPACT: NA.
TISSUE DYNAMICS IMAGING TO PREDICT DRUG THERAPEUTIC RESPONSE IN CANCER PATIENTS
Turek JJ1, An R1, Matei DE2, Nolte DD1 1Purdue University, West Lafayette, IN, USA; 2Indiana University School of Medicine, Indianapolis, IN, USA
OBJECTIVES/SPECIFIC AIMS: The goal of the research is to determine if a new biodynamic imaging technology can be used to predict therapeutic response in cancer patients by analyzing tissue drug response in tumor biopsies. METHODS/STUDY POPULATION: The tissue models were tumor spheroids (rat osteogenic sarcoma UMR-106, human colon adenocarcinoma HT-29 and DLD-1, and human pancreatic carcinoma PaCa-2) and xenografts from nude mice (human ovarian carcinoma cell lines A2780 and A2780-CP70-resistant to cisplatin). A series of drugs and biophysical treatments were applied to the tumor spheroids to generate a library of phenotypic spectrograms. RESULTS/ANTICIPATED RESULTS: TDI provided information from a 3D tissue volume that was used to assign a viability metric in response to an applied drug. TDI is label free and drug response over time may be tracked. TDI also provided a spectral fingerprint for drug mode of action and identify differential drug effects in normoxic and hypoxic tumor zones. Sub-sampling of the spectral fingerprint solved the problem of predicting drug treatment response due to tumor heterogeneity. TDI can map the zones in a tumor biopsy that are responsive or non-responsive to a particular drug cocktail or sequence. Response mapping can provide a basis to choose a drug combination or sequence that will treat the maximum volume of tumor tissue. DISCUSSION/SIGNIFICANCE OF IMPACT: TDI can provide a rapid (4–8 hour) assessment of tissue drug response in multicellular tumor spheroids and xenografts. The tissue viablility metric providese a phenotypic profile of drug response that is unique for a patient. Importantly, TDI could be used to map heterogeneity in drug response in a patient biopsy and provide the information necessary to select a drug combination or sequence that will treat the maximum volume of tumor.
TOWARD A BETTER UNDERSTANDING OF LONELINESS IN COMMUNITY-DWELLING OLDER ADULTS
Smith JM1 1Goldfarb School of Nursing at Barnes Jewish College, St. Louis, MO, USA
OBJECTIVES/SPECIFIC AIMS: The purpose of this interpretive phenomenological study was to explore the meaning of loneliness in community-dwelling older adults and to understand their daily practices in coping with loneliness. METHODS/STUDY POPULATION: The sample consisted of 8 women and 4 men. Interviews were conducted with the 12 participants utilizing several tools, including 3 separate interview guides and the UCLA Loneliness Scale, Version 3 (Russell, 1996). RESULTS/ANTICIPATED RESULTS: A critical finding was that many participants experienced loneliness as a result of disrupted meaningful engagement, due to age-related changes, as well as other losses, including death of spouse, retirement, and giving up the car. Two paradigm cases and themes representing the loneliness and coping experience emerged. Participant coping practices with loneliness included reaching out to others, helping those in need, and seeking companionship with pets. DISCUSSION/SIGNIFICANCE OF IMPACT: Many older adults are at risk for loneliness because of declining health and other age-related losses that prevent them from remaining engaged in meaningful relationships. Health care professionals can screen for loneliness to identify those at risk and can intervene to help older adults maintain connections. Recommendations for those caring for lonely older adults include active listening, vision and hearing screenings, transportation needs, pet therapy, volunteering, and engagement in social activities.
URINARY RENIN AND SOLUBLE PRORENIN RECEPTOR: NOVEL POTENTIAL BIOMARKERS OF INTRARENAL RENIN-ANGIOTENSIN SYSTEM (RAS) STATUS IN TYPE 2 DIABETIC PATIENTS
Prieto MC1, Arita DY1, Luffman C1, Bourgeois CT1, Thethi T2 1Dept. of Physiology, Tulane University School of Medicine, New Orleans, LA, USA; 2Dept. of Medicine. Tulane University HSC, New Orleans, LA, USA
OBJECTIVES/SPECIFIC AIMS: Diabetic patients exhibit a “paradoxical” low plasma renin activity (PRA) but increased intrarenal RAS activation and angiotensin (Ang)II formation. Although, they display low-to-normal PRA, their urine contains high levels of active renin but low levels of prorenin. The prorenin receptor (PRR) increases renin activity and activates prorenin. This receptor is expressed in the collecting ducts, which are the major source of prorenin in diabetes. In AngII hypertensive rats there is augmentation of soluble PRR (sPRR) in renal medulla and urine due to a transition from the full-length PRR to sPRR. The prorenin interaction with PRR provides a novel mechanism to increase intrarenal/intratubular renin activity and AngII. METHODS/STUDY POPULATION: We measured in urine, renin activity (uRen) and sPRR concentration in: Healthy (CT; N = 20); Type-2 diabetic (DM; N = 20), and Type-2 diabetic/hypertensive (DM+HTN; N = 20) patients, using RIA and ELISA, respectively. RESULTS/ANTICIPATED RESULTS: Levels of uRen, corrected by uCre, were higher in DM and DM+HTN compared to CT patients (DM: 130 ± 14; DM+HTN: 162 ± 31 vs. CT: 10 ± 1.3 ng AngI/mL/hr). Plasma sPRR levels were augmented in DM compared to CT and DM+HTN (DM: 9849 ± 828 vs. CT: 7310 ± 715 and DM+HTN: 7121 ± 373 pg/mL). In contrast, in the urine of DM and DM+HTN patients, sPRR levels were decreased (DM: 576.5 ± 130.9; DM+HTN:486.1 ± 81.64 vs. CT:1248.0 ± 345.6 pg/mg). DISCUSSION/SIGNIFICANCE OF IMPACT: These data indicate that uRen could be a potential biomarker of intrarenal RAS activation in type 2 diabetic patients. The lower urinary levels of soluble PRR in DM and DM+HTN patients suggest that membrane-bound PRR on collecting duct play a predominant role in the enhancement of uRen activity.
USING VOLATILE ORGANIC COMPOUND ANALYSIS TO IDENTIFY VIRUS-INFECTED AIRWAY CELLS
Schivo M1,2, Aksenov A3, Linderholm A2, Pasamontes A3, Peirano D3, Harper R1,2, Davis C3 1UCD School of Med, Sacramento, CA, USA; 2UCD Ctr for Comp Respir Biol & Med, Davis, CA, USA; 3UCD Dept Mech & Aero Eng, Davis, CA, USA
OBJECTIVES/SPECIFIC AIMS: Viral respiratory infections (VRI) cause morbidity in asthma and COPD. Early diagnosis may improve outcomes. Current diagnostics for VRIs are limited. Volatile organic compound (VOC) analysis of exhaled human breath may enable VRI diagnosis. The aim here is to identify if VOCs emitted from bronchial epithelial cells (BEC) can distinguish between pre- and post-human rhinovirus (HRV) infection. METHODS/STUDY POPULATION: Primary BECs came from bronchoscopy and were grown to confluence. BECs from the same subject were studied—one set uninfected and another HRV-infected. The BECs were placed into an enclosed vial where VOCs equilibrated in the headspace. VOCs were sampled using solid-phase microextraction fibers from control vials, un-infected BECs, and HRV-infected BECs; these were analyzed using gas chromatography / mass spectrometry. VOCs were identified. Data were analyzed using t-test, principle component, and partial least squares analyses. RESULTS/ANTICIPATED RESULTS: Chromatograms from native and HRV-infected BECs were compared and peak differences were visually seen. Specifically, it appears that several VOCs were expressed at higher abundance after HRV-infection. Individual VOCs identified include hexane, 2,4-dimethylhexanal, and other long-chain hydrocarbons. These VOCs were reproducible between experiments on the same subject's cells. Further testing will demonstrate inter-subject reproducibility. DISCUSSION/SIGNIFICANCE OF IMPACT: We see different expressions of VOCs between uninfected and HRV-infected BECs; we can use this difference to identify infected cells. The results are reproducible. This method can apply to other respiratory viral pathogens such as influenza. Future studies will need to test this method to identify viral infections in clinical populations.