OBJECTIVES/SPECIFIC AIMS: This study aims to first describe the unique cytokine profile and TGFbeta levels of young children with CF, then understand the pathologic effects of TGFbeta on lung function in a CF animal model. These powerful translational studies linking observations in clinical disease with a transgenic mouse model allow us a unique opportunity to investigate the role of TGFbeta in early CF lung disease. METHODS/STUDY POPULATION: Cytokine levels (TGFbeta, TNFalpha, IL-8, IL-6, HNE, and IL-1beta) in bronchoalveolar lavage fluid (BALF) from CF patients (n = 15) and non-CF control patients (n = 21) under 6 years old were determined by ELISA and Luminex assay. Tracheotomized patients without significant underlying lung disease were chosen as non-CF inflamed control patients, as they had similar levels of neutrophilic inflammation and infection as CF patients. The percentage of BAL neutrophils (% PMNs) in each sample was assessed. The relationships between cytokines were analyzed using linear regression and principal components analysis. In animal studies, CF and non-CF mice (n = 4-5 per group) were treated with intratracheal adenoviral TGFbeta1 vector, an empty vector control, or PBS. After one week, animals were collected; lung function, response to the bronchoconstrictor methacholine, and rescue with albuterol were measured utilizing a FlexiVent machine. Lungs were collected for histology. Immunohistochemistry for alpha-SMA was performed and pictures of all cross-sectional airways were obtained. Burden of ASM was assessed by dividing the square root of alpha-SMA stained airway smooth muscle by the basement membrane perimeter length of each airway. RESULTS/ANTICIPATED RESULTS: Patient characteristics of CF and non-CF inflamed control patients were similar in terms of age (3.6 yrs vs 3.3 yrs respectively, p = 0.49), positive BAL culture (13% vs 14%, p = 0.94), and % PMNs (65% vs 56%, p = 0.64). Despite these similarities, TGFbeta levels were 2-fold higher in CF versus non-CF BAL (p = 0.034). Analysis of BAL cytokines from both patient groups showed that three principal components describe 86% of total variance across the cytokine variables. These components represent different contributions from the cytokines, with TGFbeta, IL6, and % PMNs comprising one component of similarly regulated inflammatory markers. These components can distinguish CF versus non-CF patients with 77% accuracy (area under the curve: 0.77). TGFbeta concentrations were uniquely associated with increased IL-6 in CF samples (r = 0.74; p = 0.0015) but did not demonstrate association with other cytokines. After TGFbeta exposure, CF mice demonstrated greater abnormalities in airway resistance than non-CF mice, with heightened response to methacholine. Importantly, this increase in airway obstruction in CF mice was reversible with albuterol treatment, indicating airway smooth muscle dysfunction as a principal driver of lung function abnormalities. Furthermore, TGFbeta induced an increased ASM burden on lung histology in both CF and non-CF mice (p<0.05). IL-6 levels in the BAL of CF mice showed greater increases after TGFbeta treatment compared to non-CF mice (p<0.05). Empty vector control treatment did not cause lung pathology. DISCUSSION/SIGNIFICANCE OF IMPACT: Young children with CF have a unique pattern of pulmonary inflammation compared to inflamed non-CF control patients. In CF, TGFbeta pulmonary levels are uniquely associated with IL-6, a driver of ASM dysfunction in other pulmonary diseases. We followed up this clinical observation study by investigating the effect of TGFbeta on pulmonary disease in a mouse model. CF mice demonstrate increased pulmonary IL-6, airway obstruction, and ASM dysfunction after TGFbeta exposure. This study provides evidence that TGFbeta is associated with a distinct cytokine pattern that may promote ASM dysfunction in early CF lung disease. Understanding the mechanism of early CF pathophysiology will be critical in developing targeted therapeutics that can prevent early lung damage.

Genetic predispositions play an important role in the development of internalizing and externalizing behaviors. Understanding the mechanisms through which genetic risk unfolds to influence these developmental outcomes is critical for developing prevention and intervention efforts, capturing key elements of Irv's research agenda and scientific legacy. In this study, we examined the role of parenting and personality in mediating the effect of genetic risk on adolescents’ major depressive disorder and conduct disorder symptoms. Longitudinal data were drawn from a sample of 709 European American adolescents and their mothers from the Collaborative Studies on Genetics of Alcoholism. Results from multivariate path analysis indicated that adolescents’ depressive symptoms genome-wide polygenic scores (DS_GPS) predicted lower parental knowledge, which in turn was associated with more subsequent major depressive disorder and conduct disorder symptoms. Adolescents’ DS_GPS also had indirect effects on these outcomes via personality, with a mediating effect via agreeableness but not via other dimensions of personality. Findings revealed that the pattern of associations was similar across adolescent gender. Our findings emphasize the important role of evocative gene–environment correlation processes and intermediate phenotypes in the pathways of risk from genetic predispositions to complex adolescent outcomes.

OBJECTIVES/SPECIFIC AIMS: (1) Identify current barriers to coordinated care between behavior consultation and PCIT services. (2) Identify current facilitators to coordinated care between behavior consultation and PCIT services. (3) Utilize this knowledge to create and pilot a coordinated care model that will enhance PCIT and behavior consultation service outcomes. METHODS/STUDY POPULATION: Objectives 1 and 2: Two focus groups consisting of 8–10 behavior consultants will be conducted to gather initial information on barriers and facilitators to coordinated care. Participants will be recruited from the state-funded behavior consultation team, to represent consultation occurring in rural and urban settings. All focus groups will be recorded and transcribed to capture questions and comments. Focus groups will be provided with an initial 10-minute overview of PCIT, including theory, prescribed strategies, and mode of intervention. A grand tour question will then be asked to elicit consultant perceptions of PCIT (e.g., “What are your thoughts on the compatibility between PCIT and behavior consultation services”), followed by probe questions deigned to elicit more detailed information about any perceived differences based on philosophical approach; differences in what is recommended in childcare settings Versus at home, etc.; and perceived barriers to coordinated care between school and outpatient services (e.g., “What factors make coordinating care with outpatient providers challenging?). Participants will be asked about their willingness to participate in a second focus group to review materials created to enhance coordinated care, based on their feedback. Objective 3. Based on feedback from the focus groups and quantitative data regarding factors associated with PCIT outcomes, we will develop an enhanced childcare component(s) for eventual implementation. To confirm our approach, we will invite the members of both focus groups back for a second session, in which we provide them with the created materials and elicit their feedback. We will start with a grand tour question (e.g., “How do you think parents and teachers would react to these materials?”) and then follow-up with probe questions related to feasibility (e.g., “How do you anticipate using these tools?”), appropriateness (e.g., “How adequately do you feel these materials address concerns with coordinated care?”), and acceptability (e.g., “How likely are you to begin using these tools within your consultation?”). Both focus groups will be recorded and transcribed to capture questions and comments. RESULTS/ANTICIPATED RESULTS: (1) Barriers and facilitators to coordinated care will include individual (e.g., acceptability of PCIT framework) and system-level factors (e.g., ease of communication between providers). (2) There will be significant overlap in coordination between the first phase of PCIT (which focuses on positive parenting strategies) and what is prescribed by behavior consultants. (3) There will be less compatibility between the second phase of PCIT (which focuses on disciplinary strategies) and what is prescribed by behavior consultants. (4) A coordinated are model will be rated as more feasible, appropriate, and acceptable to behavior consultants than PCIT services as currently prescribed. DISCUSSION/SIGNIFICANCE OF IMPACT: Childhood disruptive behaviors are among the most frequent reasons for referral to outpatient child/adolescent mental health clinics (Sukhodolsky et al., 2016). Disruptive and aggressive behaviors are problematic, not only for victims of children who are aggressive but also for aggressive children as they age. Although effective treatments exist, families are often provided with conflicting strategies for behavior management by outpatient clinicians and behavior consultants in the daycare setting, thus providing children inconsistent feedback which will delay their attainment of new skills. These data will provide the initial foundation for the development of a coordinated care model that promotes treatment efficacy by improving the compatibility between clinic-based PCIT and daycare-based behavior consultation services.

OBJECTIVES/SPECIFIC AIMS: Transforming growth factor-beta (TGFβ) is a genetic modifier of cystic fibrosis (CF) lung disease. TGFβ’s pulmonary levels in young CF patients and its mechanism of action in CF are unknown. We examined TGFβ levels in children with CF and investigated responses of human airway epithelial cells (AECs) and mice to TGFβ. METHODS/STUDY POPULATION: TGFβ levels in bronchoalveolar lavage fluid from CF patients (n=15) and non-CF control patients (n=21)<6 years old were determined by ELISA. CF mice and non-CF mice were intratracheally treated with an adenoviral TGFβ1 vector or PBS; lungs were collected for analysis at day 7. Human CF and non-CF AECs were treated with TGFβ or PBS for 24 hours then collected for analysis. RESULTS/ANTICIPATED RESULTS: Young CF patients had higher bronchoalveolar lavage fluid TGFβ than non-CF controls (p=0.03). Mouse lungs exposed to TGFβ demonstrated inflammation, goblet cell hyperplasia, and decreased CFTR expression. CF mice had greater TGFβ-induced lung mechanics abnormalities than controls; both CF human AECs and CF mice showed higher TGFβ induced MAPK and PI3K signaling compared with controls. DISCUSSION/SIGNIFICANCE OF IMPACT: For the first time, we show increased TGFβ levels very early in CF. TGFβ drives CF lung abnormalities in mouse and human models; CF models are more sensitive to TGFβ’s effects. Understanding the role of TGFβ in promoting CF lung disease is critical to developing patient specific treatments.

Crop yields can be similar in organic and conventional systems even when weed biomass is greater in organic systems. Greater weed tolerance in organic systems may be due to differences in management-driven soil fertility properties. The goal of this experiment was to determine whether soil collected from a long-term organic cropping system with a diverse crop rotation and organic fertility inputs would support higher soil nitrogen (N) resource partitioning, as indicated by overyielding of corn–weed mixtures, than a cropping system with a less diverse crop rotation and inorganic N inputs. A replacement series greenhouse experiment was conducted using corn : smooth pigweed and corn : giant foxtail proportions of 0 : 1, 0.25 : 0.75, 0.5 : 0.5, 0.75 : 0.25, and 1 : 0 and harvested at 29, 40, or 48 d after experiment initiation (DAI). The monoculture density of corn was 4 plants pot−1 and the monoculture density of each weed species was 36 plants pot−1. Corn was consistently more competitive than both weed species at 40 and 48 DAI when soil inorganic N was limiting to growth. Corn–smooth pigweed mixtures had greater shoot biomass and shoot N content than expected based on the shoot biomass and shoot N content of monocultures (i.e., overyielding) at the onset of soil inorganic N limitation, providing some evidence for N resource partitioning. However, soil management effects on overyielding were infrequent and inconsistent among harvest dates and corn–weed mixtures, leading us to conclude that management-driven soil fertility properties did not affect corn–weed N resource partitioning during the early stages of corn growth.

Context: The detection and replication of genes involved in psychiatric outcome has been notoriously difficult. Phenotypic measurement has been offered as one explanation, although most of this discussion has focused on problems with binary diagnoses. Objective: This article focuses on two additional components of phenotypic measurement that deserve further consideration in evaluating genetic associations: (1) the measure used to reflect the outcome of interest, and (2) the developmental stage of the study population. We focus our discussion of these issues around the construct of impulsivity and externalizing disorders, and the association of these measures with a specific gene, GABRA2. Design, Setting, and Participants: Data were analyzed from the Collaborative Study on the Genetics of Alcoholism Phase IV assessment of adolescents and young adults (ages 12–26; N = 2,128). Main Outcome Measures: Alcohol dependence, illicit drug dependence, childhood conduct disorder, and adult antisocial personality disorder symptoms were measured by psychiatric interview; Achenbach youth/adult self-report externalizing scale; Zuckerman Sensation-Seeking scale; Barratt Impulsivity scale; NEO extraversion and consciousness. Results: GABRA2 was associated with subclinical levels of externalizing behavior as measured by the Achenbach in both the adolescent and young adult samples. Contrary to previous associations in adult samples, it was not associated with clinical-level DSM symptom counts of any externalizing disorders in these younger samples. There was also association with sensation-seeking and extraversion, but only in the adolescent sample. There was no association with the Barratt impulsivity scale or conscientiousness. Conclusions: Our results suggest that the pathway by which GABRA2 initially confers risk for eventual alcohol problems begins with a predisposition to sensation-seeking early in adolescence. The findings support the heterogeneous nature of impulsivity and demonstrate that both the measure used to assess a construct of interest and the age of the participants can have profound implications for the detection of genetic associations.

The discovery of a pulsar or pulsars orbiting near the Galactic Center (GC) could offer an unprecedented probe of strong-field gravity, the properties of our galaxy's supermassive black hole and insights into the paradoxical star formation history of the region. However, searching for pulsars near the GC is severely hampered by the large electron densities along our line of sight and the scattering-induced pulse broadening of the pulsar emission observed through it. As the broadened pulse length approaches the pulsar period, the periodicity in pulsar emission becomes nearly undetectable. Searches extended to higher frequencies, in an effort to reduce scattering, suffer from reduced intrinsic flux, higher system temperatures and increased atmospheric opacity. We are currently attempting to mitigate the challenges associated with searching for pulsars near the GC by employing new wide bandwidth receivers, upgraded IF distribution systems and novel digital spectrometers in a GC pulsar search campaign at the Green Bank Telescope in West Virginia, USA.

Our search will cover two frequency bands, from 12-15 GHz (Ku Band) and 18-26 GHz (K Band), during a total of approximately 30 hours of observations, with expected characteristic 10-sigma sensitivities between 5-10 micro-Jy. Our first observations are scheduled for mid-March 2012. Here we will present the status of our observations and initial results.