APPLICATIONS OF ANTIOXIDANT AND ANTI-INFLAMMATORY POLYMERS TO INHIBIT INJURY AND DISEASE

There is an undeniable link between oxidative stress, inflammation, and disease. Currently, approaches using antioxidant therapies have been largely unsuccessful due to poor delivery and bioavailability. Responding to these limitations, we have developed classes of polymer and delivery systems that can overcome the challenges of antioxidant and anti-inflammatory therapy. In our initial studies, nanoparticles of poly(trolox), a polymeric form of trolox, were surface-modified with antibodies. This modification allows for specific targeting to endothelial cells, affording controllable and localized protection against oxidative stress. We have shown these targeted nanoparticles bind, internalize, and provide protection against oxidative stress generation and cytotoxicity from iron oxide nanoparticles. In a similar fashion, we have tested the ability of poly(trolox) to prevent rheumatoid arthritis in vivo. Poly(trolox) nanoparticles were encapsulated in a PEGylated polymer to enhance circulation and biocompatibility. These particles were shown to accumulate in inflamed joint tissue, recover natural antioxidant function, suppress protein oxidation, and inhibit inflammatory markers. Lastly, we developed a class of polyphenolic compounds utilizing a non-free radical based reaction chemistry of poly(β-amino esters). The polyphenol apigenin was investigated for its anti-inflammatory properties to inhibit inflammation-mediated tumor cell metastasis. PEGylated nanoparticles that incorporated apigenin poly(β-amino ester) were developed and found to retain their anti-inflammatory efficacy while providing a long term release profile. These inhibited the ability of tumor cells to adhere to inflamed vascular cells. We also have shown that these polymers can suppress markers of inflammation responsible in enhancing tumor cell adhesion

Binding, Transcytosis and Biodistribution of Anti-PECAM-1 Iron Oxide Nanoparticles for Brain-Targeted Delivery

OBJECTIVE: Characterize the flux of platelet-endothelial cell adhesion molecule (PECAM-1) antibody-coated superparamagnetic iron oxide nanoparticles (IONPs) across the blood-brain barrier (BBB) and its biodistribution in vitro and in vivo. METHODS: Anti-PECAM-1 IONPs and IgG IONPs were prepared and characterized in house. The binding affinity of these nanoparticles was investigated using human cortical microvascular endothelial cells (hCMEC/D3). Flux assays were performed using a hCMEC/D3 BBB model. To test their immunospecificity index and biodistribution, nanoparticles were given to Sprague Dawley rats by intra-carotid infusion. The capillary depletion method was used to elucidate their distribution between the BBB and brain parenchyma. RESULTS: Anti-PECAM-1 IONPs were ~130 nm. The extent of nanoparticle antibody surface coverage was 63.6 ± 8.4%. Only 6.39 ± 1.22% of labeled antibody dissociated from IONPs in heparin-treated whole blood over 4 h. The binding affinity of PECAM-1 antibody (KD) was 32 nM with a maximal binding (Bmax) of 17 × 10(5) antibody molecules/cell. Anti-PECAM-1 IONP flux across a hCMEC/D3 monolayer was significantly higher than IgG IONP\u27s with 31% of anti-PECAM-1 IONPs in the receiving chamber after 6 h. Anti-PECAM-1 IONPs showed higher concentrations in lung and brain, but not liver or spleen, than IgG IONPs after infusion. The capillary depletion method showed that 17±12% of the anti-PECAM-1 IONPs crossed the BBB into the brain ten minutes after infusion. CONCLUSIONS: PECAM-1 antibody coating significantly increased IONP flux across the hCMEC/D3 monolayer. In vivo results showed that the PECAM-1 antibody enhanced BBB association and brain parenchymal accumulation of IONPs compared to IgG. This research demonstrates the benefit of anti-PECAM-1 IONPs for association and flux across the BBB into the brain in relation to its biodistribution in peripheral organs. The results provide insight into potential application and toxicity concerns of anti-PECAM-1 IONPs in the central nervous system

Compounds and Methods for Reducing Oxidative Stress

Antioxidant polymeric compounds are provided that comprise a plurality of monomeric portions, where each monomeric portion includes an antioxidant molecule interposed between at least two acrylate molecules, and where at least one acrylate molecule of each monomeric portion is linked by a diamine molecule to an acrylate molecule of an adjacent monomeric portion to thereby form the polymer. Methods of synthesizing polymeric compounds and methods of using the antioxidant polymeric compounds to reduce oxidative stress are also provided

Nkx2-5 and Sarcospan genetically interact in the development of the muscular ventricular septum of the heart

The muscular ventricular septum separates the flow of oxygenated and de-oxygenated blood in air-breathing vertebrates. Defects within it, termed muscular ventricular septal defects (VSDs), are common, yet less is known about how they arise than rarer heart defects. Mutations of the cardiac transcription factor NKX2-5 cause cardiac malformations, including muscular VSDs. We describe here a genetic interaction between Nkx2-5 and Sarcospan (Sspn) that affects the risk of muscular VSD in mice. Sspn encodes a protein in the dystrophin-glycoprotein complex. Sspn knockout (Sspn(KO)) mice do not have heart defects, but Nkx2-5(+/−)/Sspn(KO) mutants have a higher incidence of muscular VSD than Nkx2-5(+/−) mice. Myofibers in the ventricular septum follow a stereotypical pattern that is disrupted around a muscular VSD. Subendocardial myofibers normally run in parallel along the left ventricular outflow tract, but in the Nkx2-5(+/−)/Sspn(KO) mutant they commonly deviate into the septum even in the absence of a muscular VSD. Thus, Nkx2-5 and Sspn act in a pathway that affects the alignment of myofibers during the development of the ventricular septum. The malalignment may be a consequence of a defect in the coalescence of trabeculae into the developing ventricular septum, which has been hypothesized to be the mechanistic basis of muscular VSDs

Revised Stellar Properties of Kepler Targets for the Q1-17 (DR25) Transit Detection Run

The determination of exoplanet properties and occurrence rates using Kepler data critically depends on our knowledge of the fundamental properties (such as temperature, radius and mass) of the observed stars. We present revised stellar properties for 197,096 Kepler targets observed between Quarters 1-17 (Q1-17), which were used for the final transiting planet search run by the Kepler Mission (Data Release 25, DR25). Similar to the Q1--16 catalog by Huber et al. the classifications are based on conditioning published atmospheric parameters on a grid of Dartmouth isochrones, with significant improvements in the adopted methodology and over 29,000 new sources for temperatures, surface gravities or metallicities. In addition to fundamental stellar properties the new catalog also includes distances and extinctions, and we provide posterior samples for each stellar parameter of each star. Typical uncertainties are ~27% in radius, ~17% in mass, and ~51% in density, which is somewhat smaller than previous catalogs due to the larger number of improved logg constraints and the inclusion of isochrone weighting when deriving stellar posterior distributions. On average, the catalog includes a significantly larger number of evolved solar-type stars, with an increase of 43.5% in the number of subgiants. We discuss the overall changes of radii and masses of Kepler targets as a function of spectral type, with particular focus on exoplanet host stars.Comment: 19 pages, 13 figures. ApJS in pres

The Kepler Follow-up Observation Program

The Kepler Mission was launched on March 6, 2009 to perform a photometric survey of more than 100,000 dwarf stars to search for terrestrial-size planets with the transit technique. Follow-up observations of planetary candidates identified by detection of transit-like events are needed both for identification of astrophysical phenomena that mimic planetary transits and for characterization of the true planets and planetary systems found by Kepler. We have developed techniques and protocols for detection of false planetary transits and are currently conducting observations on 177 Kepler targets that have been selected for follow-up. A preliminary estimate indicates that between 24% and 62% of planetary candidates selected for follow-up will turn out to be true planets.Comment: 12 pages, submitted to the Astrophysical Journal Letter

The Composition of Comets

This paper is the result of the International Cometary Workshop, held in Toulouse, France in April 2014, where the participants came together to assess our knowledge of comets prior to the ESA Rosetta Mission. In this paper, we look at the composition of the gas and dust from the comae of comets. With the gas, we cover the various taxonomic studies that have broken comets into groups and compare what is seen at all wavelengths. We also discuss what has been learned from mass spectrometers during flybys. A few caveats for our interpretation are discussed. With dust, much of our information comes from flybys. They include {\it in situ} analyses as well as samples returned to Earth for laboratory measurements. Remote sensing IR observations and polarimetry are also discussed. For both gas and dust, we discuss what instruments the Rosetta spacecraft and Philae lander will bring to bear to improve our understanding of comet 67P/Churyumov-Gerasimenko as "ground-truth" for our previous comprehensive studies. Finally, we summarize some of the initial Rosetta Mission findings.Comment: To appear in Space Science Review

GATA6 modulates the ductular reaction to bile duct ligation

Background GATA6, a transcription factor expressed in cholangiocytes, has been implicated in the response to liver injury. In biliary atresia, a disease characterized by extrahepatic bile duct obstruction, liver expression of GATA6 increases with pathological bile duct expansion and decreases after successful Kasai portoenterostomy. The aim of this study was to garner genetic evidence that GATA6 is involved in ductular formation/expansion. Methods The murine Gata6 gene was conditionally deleted using Alb-cre, a transgene expressed in hepatoblasts (the precursors of hepatocytes and cholangiocytes) and mature hepatocytes. Bile duct ligation (BDL) was used to model biliary obstruction. Results Alb-Cre;Gata6(flox/flox) mice were viable and fertile. Cre-mediated recombination of Gata6 in hepatocytes had little impact on cellular structure or function. GATA6 immunoreactivity was retained in a majority of biliary epithelial cells in adult Alb-Cre;Gata6(flox/flox) mice, implying that surviving cholangiocytes were derived from hepatoblasts that had escaped biallelic Cre-mediated recombination. Although GATA6 immunoreactivity was preserved in cholangiocytes, Alb-cre;Gata6(flox/flox) mice had a demonstrable biliary phenotype. A neutrophil-rich infiltrate surrounded newly formed bile ducts in neonatal Alb-Cre;Gata6(flox/flox) mice. Foci of fibrosis/necrosis, presumed to reflect patchy defects in bile duct formation, were observed in the livers of 37% of adult Alb-cre;Gata6(flox/flox) mice and 0% of controls (p <0.05). Most notably, Alb-cre;Gata6(flox/flox) mice had an altered response to BDL manifest as reduced survival, impaired bile ductule proliferation, increased parenchymal necrosis, reduced fibrosis, and enhanced macrophage accumulation in the portal space. Conclusions GATA6 orchestrates intrahepatic biliary remodeling and mitigates liver injury following extrahepatic bile duct obstruction. Graphic abstractPeer reviewe

Kepler-4B: A Hot Neptune-Like Planet of A G0 Star Near Main-Sequence Turnoff

Early time-series photometry from NASA's Kepler spacecraft has revealed a planet transiting the star we term Kepler-4, at R.A. = 19(h)02(m)27.(s)68, delta = +50 degrees 08'08 '' 7. The planet has an orbital period of 3.213 days and shows transits with a relative depth of 0.87 x 10(-3) and a duration of about 3.95 hr. Radial velocity (RV) measurements from the Keck High Resolution Echelle Spectrometer show a reflex Doppler signal of 9.3(-1.9)(+1.1) m s(-1), consistent with a low-eccentricity orbit with the phase expected from the transits. Various tests show no evidence for any companion star near enough to affect the light curve or the RVs for this system. From a transit-based estimate of the host star's mean density, combined with analysis of high-resolution spectra, we infer that the host star is near turnoff from the main sequence, with estimated mass and radius of 1.223(-0.091)(+0.053) M(circle dot) and 1.487(-0.084)(+0.071) R(circle dot).We estimate the planet mass and radius to be {M(P), R(P)} = {24.5 +/- 3.8 M(circle plus), 3.99 +/- 0.21 R(circle plus)}. The planet's density is near 1.9 g cm(-3); it is thus slightly denser and more massive than Neptune, but about the same size.W. M. Keck FoundationNASA's Science Mission DirectorateAstronom

Kepler Observations of Transiting Hot Compact Objects

Kepler photometry has revealed two unusual transiting companions orbiting an early A-star and a late B-star. In both cases the occultation of the companion is deeper than the transit. The occultation and transit with follow-up optical spectroscopy reveal a 9400 K early A-star, KOI-74 (KIC 6889235), with a companion in a 5.2 day orbit with a radius of 0.08 Rsun and a 10000 K late B-star KOI-81 (KIC 8823868) that has a companion in a 24 day orbit with a radius of 0.2 Rsun. We infer a temperature of 12250 K for KOI-74b and 13500 K for KOI-81b. We present 43 days of high duty cycle, 30 minute cadence photometry, with models demonstrating the intriguing properties of these object, and speculate on their nature.Comment: 12 pages, 3 figures, submitted to ApJL (updated to correct KOI74 lightcurve