OBJECTIVES/GOALS: Lack of blood vessels remains a major obstacle in tissue regeneration. Angiopoietin 1 and 2 modulate angiogenesis through the Tie2 receptor tyrosine kinase. Ang1 activates pAKT to promote endothelial cell survival while Ang2 antagonizes these effects. We aim to dissect the Ang/Tie2 pathway to uncover the molecular basis for these opposing effects. METHODS/STUDY POPULATION: Ang1 and Ang2 bind Tie2 via nearly identical F-domains (Fd). To investigate the molecular basis regulating the Tie2 pathway, we generated a series of computationally designed self-assembling protein scaffolds presenting F-domains in a wide range of valencies and geometries using Rosette Molecular Modeling Suite. We examined the protein kinase activation, cell migration, and blood vessel formation produced by the designed proteins in human umbilical vein endothelial cells. RESULTS/ANTICIPATED RESULTS: Two phenotypic classes were demonstrated by the number of presented F domains: scaffolds presenting 3 or 4 Fd have Ang2 like activity, upregulating pFAK and pERK but not pAKT and failing to induce cell migration and tube formation; scaffolds presenting more than 6 Fd have Ang1 like activity, upregulating the three signaling branches and enhancing cell migration and tube formation. Scaffolds with 8 or more Fd show superagonist activity, producing significantly stronger phenotypes than Ang1. These results suggest that Fd valency largely determines Ang1 vs Ang2 signaling outcomes, and our designed superagonists can outperform Ang1 in vascularization and wound healing. In in vivo experiments, nanoparticles displaying 60 copies of Fd produce significant revascularization in hemorrhagic brains. DISCUSSION/SIGNIFICANCE OF IMPACT: Targeting the Tie2 pathway is a new paradigm in regenerative medicine. Our designed constructs will enable us to generate high-affinity Tie2 agonists and antagonists as drugs to control angiogenesis, enabling tissue regeneration that recapitulates the biological architecture of the native tissue physiology, improving organ transplant outcome.

The Inner Mongolian Plateau lies along the northern limit reached by the East Asian summer monsoon. This geographic setting makes it especially sensitive to environmental change and an excellent site for understanding Quaternary East Asian monsoon variability. In this study we present new results of hydrogen isotopic compositions of fatty acids extracted from sediments, which were used to construct Holocene paleoprecipitation (or moisture) changes in Northern China. The hydrogen isotopic composition (D/H ratio) of n-acids in the sedimentary sequence of the Duoerji peat, Inner Mongolia, was determined with gas chromatography and mass spectrometry. Changes in the precipitation from middle Inner Mongolia are recorded by the D/H ratio of n-C20, n-C22, n-C24, n-C26, n-C28 acids (δD). From 10–9 ka, the relatively high δD values indicate reduced precipitation in the Early Holocene. Subsequently, increased precipitation is reflected by reduced δD values from 9–5.5 ka. After 5.5 ka, gradually increasing δD values record an overall decrease in precipitation. The precipitation trends established for the Duoerji sequence are consistent with other major paleoclimate proxies in the East Asian monsoon region, especially with a distinct Holocene optimum of increased monsoonal activity from 9–5.5 ka. The δD resulting paleo-precipitation record clearly shows that the Holocene climate in Northern China is basically controlled by the insolation changes.

The role that vitamin D plays in pulmonary function remains uncertain. Epidemiological studies reported mixed findings for serum 25-hydroxyvitamin D (25(OH)D)–pulmonary function association. We conducted the largest cross-sectional meta-analysis of the 25(OH)D–pulmonary function association to date, based on nine European ancestry (EA) cohorts (n 22 838) and five African ancestry (AA) cohorts (n 4290) in the Cohorts for Heart and Aging Research in Genomic Epidemiology Consortium. Data were analysed using linear models by cohort and ancestry. Effect modification by smoking status (current/former/never) was tested. Results were combined using fixed-effects meta-analysis. Mean serum 25(OH)D was 68 (sd 29) nmol/l for EA and 49 (sd 21) nmol/l for AA. For each 1 nmol/l higher 25(OH)D, forced expiratory volume in the 1st second (FEV1) was higher by 1·1 ml in EA (95 % CI 0·9, 1·3; P<0·0001) and 1·8 ml (95 % CI 1·1, 2·5; P<0·0001) in AA (Prace difference=0·06), and forced vital capacity (FVC) was higher by 1·3 ml in EA (95 % CI 1·0, 1·6; P<0·0001) and 1·5 ml (95 % CI 0·8, 2·3; P=0·0001) in AA (Prace difference=0·56). Among EA, the 25(OH)D–FVC association was stronger in smokers: per 1 nmol/l higher 25(OH)D, FVC was higher by 1·7 ml (95 % CI 1·1, 2·3) for current smokers and 1·7 ml (95 % CI 1·2, 2·1) for former smokers, compared with 0·8 ml (95 % CI 0·4, 1·2) for never smokers. In summary, the 25(OH)D associations with FEV1 and FVC were positive in both ancestries. In EA, a stronger association was observed for smokers compared with never smokers, which supports the importance of vitamin D in vulnerable populations.

Electron and proton microprobes, along with electron backscatter diffraction (EBSD) analysis were used to study the microstructure of the contemporary Al–Cu–Li alloy AA2099-T8. In electron probe microanalysis, wavelength and energy dispersive X-ray spectrometry were used in parallel with soft X-ray emission spectroscopy (SXES) to characterize the microstructure of AA2099-T8. The electron microprobe was able to identify five unique compositions for constituent intermetallic (IM) particles containing combinations of Al, Cu, Fe, Mn, and Zn. A sixth IM type was found to be rich in Ti and B (suggesting TiB2), and a seventh IM type contained Si. EBSD patterns for the five constituent IM particles containing Al, Cu, Fe, Mn, and Zn indicated that they were isomorphous with four phases in the 2xxx series aluminium alloys including Al6(Fe, Mn), Al13(Fe, Mn)4 (two slightly different compositions), Al37Cu2Fe12 and Al7Cu2Fe. SXES revealed that Li was present in some constituent IM particles. Al SXES mapping revealed an Al-enriched (i.e., Cu, Li-depleted) zone in the grain boundary network. From the EBSD analysis, the kernel average misorientation map showed higher levels of localized misorientation in this region, suggesting greater deformation or stored energy. Proton-induced X-ray emission revealed banding of the TiB2 IM particles and Cu inter-band enrichment.

This paper presents the first major data release and survey description for the ANU WiFeS SuperNovA Programme. ANU WiFeS SuperNovA Programme is an ongoing supernova spectroscopy campaign utilising the Wide Field Spectrograph on the Australian National University 2.3-m telescope. The first and primary data release of this programme (AWSNAP-DR1) releases 357 spectra of 175 unique objects collected over 82 equivalent full nights of observing from 2012 July to 2015 August. These spectra have been made publicly available via the WISEREP supernova spectroscopy repository.

We analyse the ANU WiFeS SuperNovA Programme sample of Type Ia supernova spectra, including measurements of narrow sodium absorption features afforded by the high spectral resolution of the Wide Field Spectrograph instrument. In some cases, we were able to use the integral-field nature of the Wide Field Spectrograph instrument to measure the rotation velocity of the SN host galaxy near the SN location in order to obtain precision sodium absorption velocities. We also present an extensive time series of SN 2012dn, including a near-nebular spectrum which both confirms its ‘super-Chandrasekhar’ status and enables measurement of the sub-solar host metallicity at the SN site.

Accelerator mass spectrometry (AMS) is the most sensitive method for measuring 129I in environmental samples available today, with a detection limit of about 10–15 for 129I/127I. A drawback of the technique is the time-consuming chemical separation required to prepare AMS targets from raw samples. This step significantly limits applications requiring rapid analyses and large numbers of samples, for example, in monitoring studies associated with nuclear accidents. This work introduces a direct method for 129I measurements by AMS that does not require chemical separation. In this approach, stable iodine (127I) is added to a matrix of niobium (Nb) powder and mixed with dried raw sample. This mixture is pressed directly into a sputter target for AMS analysis. Two types of environmental samples have been tested in this work, seaweed and sediment. No anomalous behavior was noted in the Cs+ sputtering behavior of the targets prepared from these materials. The 129I/127I ratios and 129I concentrations measured by this rapid method were found to be in agreement with reported values that used a conventional AMS method for the same material. Based on our findings, we expect that such rapid measurements can be applied to a wide variety of materials, in addition to seaweed and sediment, as long as the sputtering-induced adverse effects do not prevent the stable operation of the ion source. The method is especially useful for screening large numbers of samples before more precise analyses are made.

China is home to three subspecies of tiger Panthera tigris but there are no estimates of the size of any of the populations. We detected a population of the Endangered Amur tiger Panthera tigris altaica in Hunchun Nature Reserve in Jilin Province using both mitochondrial DNA and nuclear microsatellite loci. Four male and one female tigers were detected, indicating the potential for a small breeding group. However, genetic diversity was low overall, with six loci showing a heterozygote deficiency and a mean of 2.55 alleles per locus. This study is the first estimate of the wild Amur tiger population in China to use non-invasive techniques, and the presence of a female tiger indicates this is a potentially viable population. We provide baseline genetic diversity estimates to support monitoring of the population. The small number of tiger scats located indicates the importance of continuing the current conservation efforts for this tiger subspecies in Hunchun Nature Reserve. Such efforts include reducing poaching of tigers and their prey, and implementation of management plans to encourage the persistence and recovery of tigers in this area.

We measured the spin-orbit misalignment for WASP-79b, a transiting hot Jupiter from the WASP survey. Using the Rossiter-McLaughlin effect during the transit event, we determined the sky-projected obliquity to be λ = −106+10−8○. This result indicates that the planet is in a nearly polar orbit.