Coupling Strategies for the Synthesis of Peptide-Oligonucleotide Conjugates for Patterned Synthetic Biomineralization

This work describes preparation strategies for peptide-oligonucleotide conjugates that combine the self-assembling behavior of DNA oligonucleotides with the molecular recognition capabilities of peptides. The syntheses include a solution-phase fragment coupling reaction and a solid-phase fragment coupling strategy where the oligonucleotide has been immobilized on DEAE Sepharose. The yield of four coupling reagents is evaluated, two reagents in water, EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride) and DMTMM (4-(4,6-dimethoxy[1,3,5]triazin-2-yl)-4-methyl-morpholinium chloride), and two in dimethylformamide (DMF), PyBOP ((Benzotriazol-1-yloxy) tripyrrolidinophosphonium hexafluorophosphate) and HBTU (O-benzotriazole-N,N,N′,N′-tetramethyluronium hexafluorophosphate), while the oligonucleotide fragment is either in solution or immobilized on DEAE. These coupling strategies rely on an unprotected 5′ amino linker on the oligonucleotide reacting with the peptide C-terminus. The peptide, selected from a combinatorial library for its gold-binding behavior, was 12 amino acids long with an N-terminus acetyl cap. Formation of the conjugates was confirmed by gel electrophoresis and mass spectrometry while molecular recognition functionality of the peptide portion was verified using atomic force microscopy. Solution-phase yields were superior to their solid-phase counterparts. EDC resulted in the highest yield for both solution-phase (95%) and solid-phase strategies (24%), while the DMF-based reagents, PyBOP and HBTU, resulted in low yields with reduced recovery. All recoverable conjugates demonstrated gold nanoparticle templating capability

C/EBPβ-1 promotes transformation and chemoresistance in Ewing sarcoma cells.

CEBPB copy number gain in Ewing sarcoma was previously shown to be associated with worse clinical outcome compared to tumors with normal CEBPB copy number, although the mechanism was not characterized. We employed gene knockdown and rescue assays to explore the consequences of altered CEBPB gene expression in Ewing sarcoma cell lines. Knockdown of EWS-FLI1 expression led to a decrease in expression of all three C/EBPβ isoforms while re-expression of EWS-FLI1 rescued C/EBPβ expression. Overexpression of C/EBPβ-1, the largest of the three C/EBPβ isoforms, led to a significant increase in colony formation when cells were grown in soft agar compared to empty vector transduced cells. In addition, depletion of C/EBPβ decreased colony formation, and re-expression of either C/EBPβ-1 or C/EBPβ-2 rescued the phenotype. We identified the cancer stem cell marker ALDH1A1 as a target of C/EBPβ in Ewing sarcoma. Furthermore, increased expression of C/EBPβ led to resistance to chemotherapeutic agents. In summary, we have identified CEBPB as an oncogene in Ewing sarcoma. Overexpression of C/EBPβ-1 increases transformation, upregulates expression of the cancer stem cell marker ALDH1A1, and leads to chemoresistance

DNA Damage Response and Repair: Insights into Strategies for Radiation Sensitization

The incorporation of radiotherapy into multimodality treatment plans has led to significant improvements in glioma patient survival. However, local recurrence from glioma resistance to ionizing radiation remains a therapeutic challenge. The tumoricidal effect of radiation therapy is largely attributed to the induction of dsDNA breaks (DSBs). In the past decade, there have been tremendous strides in understanding the molecular mechanisms underlying DSB repair. The identification of gene products required for DSB repair has provided novel therapeutic targets. Recent studies revealed that many US FDA-approved cancer agents inhibit DSB repair by interacting with repair proteins. This article will aim to provide discussion of DSB repair mechanisms to provide molecular targets for radiation sensitization of gliomas and a discussion of FDA-approved cancer therapies that modulate DSB repair to highlight opportunities for combination therapy with radiotherapy for glioma therapy

Initial Observations of Lunar Impact Melts and Ejecta Flows with the Mini-RF Radar

The Mini-RF radar on the Lunar Reconnaissance Orbiter's spacecraft has revealed a great variety of crater ejecta flow and impact melt deposits, some of which were not observed in prior radar imaging. The craters Tycho and Glushko have long melt flows that exhibit variations in radar backscatter and circular polarization ratio along the flow. Comparison with optical imaging reveals that these changes are caused by features commonly seen in terrestrial lava flows, such as rafted plates, pressure ridges, and ponding. Small (less than 20 km) sized craters also show a large variety of features, including melt flows and ponds. Two craters have flow features that may be ejecta flows caused by entrained debris flowing across the surface rather than by melted rock. The circular polarization ratios (CPRs) of the impact melt flows are typically very high; even ponded areas have CPR values between 0.7-1.0. This high CPR suggests that deposits that appear smooth in optical imagery may be rough at centimeter- and decimeter- scales. In some places, ponds and flows are visible with no easily discernable source crater. These melt deposits may have come from oblique impacts that are capable of ejecting melted material farther downrange. They may also be associated with older, nearby craters that no longer have a radar-bright proximal ejecta blanket. The observed morphology of the lunar crater flows has implications for similar features observed on Venus. In particular, changes in backscatter along many of the ejecta flows are probably caused by features typical of lava flows

The Transit Ingress and the Tilted Orbit of the Extraordinarily Eccentric Exoplanet HD 80606b

We present the results of a transcontinental campaign to observe the 2009 June 5 transit of the exoplanet HD 80606b. We report the first detection of the transit ingress, revealing the transit duration to be 11.64 +/- 0.25 hr and allowing more robust determinations of the system parameters. Keck spectra obtained at midtransit exhibit an anomalous blueshift, giving definitive evidence that the stellar spin axis and planetary orbital axis are misaligned. The Keck data show that the projected spin-orbit angle is between 32-87 deg with 68.3% confidence and between 14-142 deg with 99.73% confidence. Thus the orbit of this planet is not only highly eccentric (e=0.93), but is also tilted away from the equatorial plane of its parent star. A large tilt had been predicted, based on the idea that the planet's eccentric orbit was caused by the Kozai mechanism. Independently of the theory, it is noteworthy that all 3 exoplanetary systems with known spin-orbit misalignments have massive planets on eccentric orbits, suggesting that those systems migrate differently than lower-mass planets on circular orbits.Comment: ApJ, in press [13 pg

KELT-10b: The First Transiting Exoplanet from the KELT-South Survey -- A Hot Sub-Jupiter Transiting a V = 10.7 Early G-Star

We report the discovery of KELT-10b, the first transiting exoplanet discovered using the KELT-South telescope. KELT-10b is a highly inflated sub-Jupiter mass planet transiting a relatively bright $V = 10.7$ star (TYC 8378-64-1), with T$_{eff}$ = $5948\pm74$ K, $\log{g}$ = $4.319_{-0.030}^{+0.020}$ and [Fe/H] = $0.09_{-0.10}^{+0.11}$, an inferred mass M$_{*}$ = $1.112_{-0.061}^{+0.055}$ M$_{\odot}$ and radius R$_{*}$ = $1.209_{-0.035}^{+0.047}$ R$_{\odot}$. The planet has a radius R$_{P}$ = $1.399_{-0.049}^{+0.069}$ R$_{J}$ and mass M$_{P}$ = $0.679_{-0.038}^{+0.039}$ M$_{J}$. The planet has an eccentricity consistent with zero and a semi-major axis $a$ = $0.05250_{-0.00097}^{+0.00086}$ AU. The best fitting linear ephemeris is $T_{0}$ = 2457066.72045$\pm$0.00027 BJD$_{TDB}$ and P = 4.1662739$\pm$0.0000063 days. This planet joins a group of highly inflated transiting exoplanets with a radius much larger and a mass much less than those of Jupiter. The planet, which boasts deep transits of 1.4%, has a relatively high equilibrium temperature of T$_{eq}$ = $1377_{-23}^{+28}$ K, assuming zero albedo and perfect heat redistribution. KELT-10b receives an estimated insolation of $0.817_{-0.054}^{+0.068}$ $\times$ 10$^9$ erg s$^{-1}$ cm$^{-2}$, which places it far above the insolation threshold above which hot Jupiters exhibit increasing amounts of radius inflation. Evolutionary analysis of the host star suggests that KELT-10b is unlikely to survive beyond the current subgiant phase, due to a concomitant in-spiral of the planet over the next $\sim$1 Gyr. The planet transits a relatively bright star and exhibits the third largest transit depth of all transiting exoplanets with V $<$ 11 in the southern hemisphere, making it a promising candidate for future atmospheric characterization studies.Comment: 20 pages, 13 figures, 7 tables, accepted for publication in MNRA

Bayesian Methods for Exoplanet Science

Exoplanet research is carried out at the limits of the capabilities of current telescopes and instruments. The studied signals are weak, and often embedded in complex systematics from instrumental, telluric, and astrophysical sources. Combining repeated observations of periodic events, simultaneous observations with multiple telescopes, different observation techniques, and existing information from theory and prior research can help to disentangle the systematics from the planetary signals, and offers synergistic advantages over analysing observations separately. Bayesian inference provides a self-consistent statistical framework that addresses both the necessity for complex systematics models, and the need to combine prior information and heterogeneous observations. This chapter offers a brief introduction to Bayesian inference in the context of exoplanet research, with focus on time series analysis, and finishes with an overview of a set of freely available programming libraries.Comment: Invited revie

Stellar Spin-Orbit Misalignment in a Multiplanet System

Stars hosting hot Jupiters are often observed to have high obliquities, whereas stars with multiple co-planar planets have been seen to have low obliquities. This has been interpreted as evidence that hot-Jupiter formation is linked to dynamical disruption, as opposed to planet migration through a protoplanetary disk. We used asteroseismology to measure a large obliquity for Kepler-56, a red giant star hosting two transiting co-planar planets. These observations show that spin-orbit misalignments are not confined to hot-Jupiter systems. Misalignments in a broader class of systems had been predicted as a consequence of torques from wide-orbiting companions, and indeed radial-velocity measurements revealed a third companion in a wide orbit in the Kepler-56 system.Comment: Accepted for publication in Science, published online on October 17 2013; PDF includes main article and supplementary materials (65 pages, 27 figures, 7 tables); v2: small correction to author lis

Values of the Federal Public Lands

v, 158 p. ; 28 cmhttps://scholar.law.colorado.edu/books_reports_studies/1038/thumbnail.jp

Obliquities of Hot Jupiter host stars: Evidence for tidal interactions and primordial misalignments

We provide evidence that the obliquities of stars with close-in giant planets were initially nearly random, and that the low obliquities that are often observed are a consequence of star-planet tidal interactions. The evidence is based on 14 new measurements of the Rossiter-McLaughlin effect (for the systems HAT-P-6, HAT-P-7, HAT-P-16, HAT-P-24, HAT-P-32, HAT-P-34, WASP-12, WASP-16, WASP-18, WASP-19, WASP-26, WASP-31, Gl 436, and Kepler-8), as well as a critical review of previous observations. The low-obliquity (well-aligned) systems are those for which the expected tidal timescale is short, and likewise the high-obliquity (misaligned and retrograde) systems are those for which the expected timescale is long. At face value, this finding indicates that the origin of hot Jupiters involves dynamical interactions like planet-planet interactions or the Kozai effect that tilt their orbits, rather than inspiraling due to interaction with a protoplanetary disk. We discuss the status of this hypothesis and the observations that are needed for a more definitive conclusion.Comment: Accepted for publication in ApJ; typos corrected, 2 broken references fixed, 26 pages, 25 figure