Alginate Hydrogels as Three-Dimensional Scaffolds for \u3ci\u3ein Vitro\u3c/i\u3e Culture Models of Growth Plate Cartilage Development and Porcine Embryo Elongation

The establishment of in vitro culture models utilizes tissue engineering principles to design functional mimics of in vivo environments in vitro. Advantages for the use of in vitro culture models include ethical alleviation of animal models for therapeutic testing, cost efficiency, and a greater ability to study specific mechanisms via a systematic, ground-up approach to development. In this thesis, alginate hydrogels are utilized in the development of in vitro culture models of porcine embryo elongation and growth plate cartilage development. First, the effect of scaffold and modifications to the scaffold were explored in both projects. In order to modulate cell-scaffold interactions, the alginate molecule was covalently modified with the arginine-glycine-aspartic acid (RGD) peptide, which has been extensively used to promote cell adhesion in vitro. In the porcine embryo elongation studies, a previously established culture system was used as a tool to encapsulate embryos and explore the effect of soluble factors on embryo survivability, rate of morphological changes, gene expression and hormone production by using RGD-modified alginate and alginate mixed with osteopontin (SPP1), a glycoprotein known to be secreted in the uterus. RGD-alginate encapsulation led to significant increases in embryo survivability and morphological changes along with corresponding cellular outputs. In the growth plate cartilage development model, chondrocytes were encapsulated in alginate and RGD-alginate to elucidate effects of cell adhesion, mimicking cell-matrix interactions within the growth plate, on chondrocyte phenotype. Additionally, alginate encapsulation was utilized to culture chondrocytes in media supplemented with parathyroid hormone (PTH) or Indian hedgehog (IHH) to begin to study the mechanisms behind these two molecules on the establishment of the zonal architecture and column formation that is unique to the growth plate. Cell cycle distribution, gene expression immunofluorescence and fluorescent in situ hybridization techniques together indicated our ability to suppress hypertrophy using alginate hydrogel scaffolds and/or signaling molecules. Overall, utilization of these in vitro culture models as tools to study developmental mechanisms can be used to advance both the fields of porcine embryology and cartilage developmental biology to improve pregnancy outcomes in pigs and to engineer therapies for the treatment of growth plate injuries and diseases. Advisor: Angela K. Pannie

SESSION 1: CREW Seattle Presentation

ABSTRACT: Is there Still a Place in Seattle for the Single-Family Detached Housing Typology, Given the Acute Need for Affordable Housing? This expert panel will explore the intersection between existing zoning laws and well-established neighborhood patterns of development, on the one hand, and the acute need for the increased production and availability of affordable housing, in the greater Seattle area, including in and near the City of Seattle’s Central Business District, as well as other close-in employment centers, on the other hand. The genesis of this Special Topic in the Innovating the Built Environment SITIE2020 course came out of a series of articles published during the SITIE2019 course reporting on several cities throughout the U.S., including Minneapolis, MN, contemplating the elimination of single-family detached zoning from their zoning and land use codes as part of a larger strategy for ramping up affordable housing production

Evidence for internal field in graphite: A conduction electron spin resonance study

We report conduction electron spin resonance measurements performed on highly oriented pyrolitic graphite samples between 10 K and 300 K using S (f = 4 GHz), X (f = 9.4 GHz), and Q (f = 34.4 GHz) microwave bands for the external dc-magnetic field applied parallel (H || c) and perpendicular (H perp c) to the sample hexagonal c-axis. The results obtained in the H || c geometry are interpreted in terms of the presence of an effective internal ferromagnetic-like field Heff-int(T,H) that increases as the temperature decreases and the applied dc-magnetic field increases. We associate the occurrence of the Heff-int(T,H) with the field-induced metal-insulator transition in graphite and discuss its origin in the light of relevant theoretical models.Comment: 10 pages (tex), 5 figures (ps

A Tunable, Three-Dimensional \u3ci\u3eIn Vitro\u3c/i\u3e Culture Model of Growth Plate Cartilage Using Alginate Hydrogel Scaffolds

Defining the final size and geometry of engineered tissues through precise control of the scalar and vector components of tissue growth is a necessary benchmark for regenerative medicine, but it has proved to be a significant challenge for tissue engineers. The growth plate cartilage that promotes elongation of the long bones is a good model system for studying morphogenetic mechanisms because cartilage is composed of a single cell type, the chondrocyte; chondrocytes are readily maintained in culture; and growth trajectory is predominately in a single vector. In this cartilage, growth is generated via a differentiation program that is spatially and temporally regulated by an interconnected network composed of long- and short-range signaling mechanisms that together result in the formation of functionally distinct cellular zones. To facilitate investigation of the mechanisms underlying anisotropic growth, we developed an in vitro model of the growth plate cartilage by using neonatal mouse growth plate chondrocytes encapsulated in alginate hydrogel beads. In bead cultures, encapsulated chondrocytes showed high viability, cartilage matrix deposition, low levels of chondrocyte hypertrophy, and a progressive increase in cell proliferation over 7 days in culture. Exogenous factors were used to test functionality of the parathyroid-related protein–Indian hedgehog (PTHrP-IHH) signaling interaction, which is a crucial feedback loop for regulation of growth. Consistent with in vivo observations, exogenous PTHrP stimulated cell proliferation and inhibited hypertrophy, whereas IHH signaling stimulated chondrocyte hypertrophy. Importantly, the treatment of alginate bead cultures with IHH or thyroxine resulted in formation of a discrete domain of hypertrophic cells that mimics tissue architecture of native growth plate cartilage. Together, these studies are the first demonstration of a tunable in vitro system to model the signaling network interactions that are required to induce zonal architecture in growth plate chondrocytes, which could also potentially be used to grow cartilage cultures of specific geometries to meet personalized patient needs

Interstellar Comets from Post-Main Sequence Systems as Tracers of Extrasolar Oort Clouds

Interstellar small bodies are unique probes into the histories of exoplanetary systems. One hypothesized class of interlopers are "Jurads," exo-comets released into the Milky Way during the post-main sequence as the thermally-pulsing asymptotic giant branch (AGB) host stars lose mass. In this study, we assess the prospects for the Legacy Survey of Space and Time (LSST) to detect a Jurad and examine whether such an interloper would be observationally distinguishable from exo-comets ejected during the (pre-)main sequence. Using analytic and numerical methods, we estimate the fraction of exo-Oort Cloud objects that are released from 1-8 solar mass stars during post-main sequence evolution. We quantify the extent to which small bodies are altered by the increased luminosity and stellar outflows during the AGB, finding that some Jurads may lack hypervolatiles and that stellar winds could deposit dust that covers the entire exo-comet surface. Next, we construct models of the interstellar small body reservoir for various size-frequency distribution slopes, characteristic sizes, and the total mass sequestered in the minor planets of exo-Oort Clouds. Even with the LSST's increased search volume compared to contemporary surveys, we find that detecting a Jurad is unlikely but not infeasible given the current understanding of (exo)planet formation.Comment: 28 pages, 13 figures; accepted to PS

Metallicities for 13 nearby open clusters from high-resolution spectroscopy of dwarf and giant stars. Stellar metallicity, stellar mass, and giant planets

We present a study of accurate stellar parameters and iron abundances for 39 giants and 16 dwarfs in the 13 open clusters IC2714, IC4651, IC4756, NGC2360, NGC2423, NGC2447 (M93), NGC2539, NGC2682 (M67), NGC3114, NGC3680, NGC4349, NGC5822, NGC6633. The analysis was done using a set of high-resolution and high-S/N spectra obtained with the UVES spectrograph (VLT). These clusters are currently being searched for planets using precise radial velocities. For all the clusters, the derived average metallicities are close to solar. Interestingly, the values derived seem to depend on the line-list used. This dependence and its implications for the study of chemical abundances in giants stars are discussed. We show that a careful choice of the lines may be crucial for the derivation of metallicities for giant stars on the same metallicity scale as those derived for dwarfs. Finally, we discuss the implications of the derived abundances for the metallicity- and mass-giant planet correlation. We conclude that a good knowledge of the two parameters is necessary to correctly disentangle their influence on the formation of giant planets.Comment: accepted for publication in A&

Trans-Gulf of Mexico loop migration of tree swallows revealed by solar geolocation

One of the greatest feats of avian migration is the non-stop crossing of extensive areas of inhospitable habitat such as deserts and seas. Differences in spring and autumn migration routes have been reported in species that cross such barriers, and are thought to have evolved in response to seasonal variation in prevailing wind direction. We tested the hypothesis that migration routes vary seasonally with respect to the Gulf of Mexico in the tree swallow Tachycineta bicolor using solar geolocators attached and retrieved at 4 breeding sites in central North America. We found that 100 % of birds (n = 10) made a trans-Gulf flight of \u3e850 km from Louisiana south to their wintering grounds in the Yucatan Peninsula in 12–36 hours, achieving minimum ground speeds as high as 32 m/s. Although most days during autumn migration were characterized by unfavorable headwinds blowing to the northwest, migration over the Gulf mostly occurred on days with strong winds blowing to the south. In contrast, in 8 of 9 (88 %) birds on spring migration returned from the wintering grounds towards Louisiana following a clockwise loop pat tern flying over land to the west around the Gulf. During this spring period there were few days with prevailing winds from the south to assist northward migration. Results suggest that, despite being up to three times further (ca. 2,700 km), a coastal cir- cum-Gulf spring migration represents the less risky route when wind conditions are not favorable. These findings also help to re solve a long-standing dispute in the literature concerning migration patterns between the US Gulf coast and Mexico, and provide insight into the factors shaping migration strategies of small songbirds migrating across large bodies of water

Kinematics, ages and metallicities for F and G type stars in the solar neighbourhood

A new metallicity distribution and an age-metallicity relation are presented for 437 nearby F and G turn-off and sub-giant stars selected from radial velocity data of Nidever et al. Photometric metallicities are derived from uvby-H\beta photometry, and the stellar ages from the isochrones of Bergbusch & VandenBerg as transformed to $uvby$ photometry using the methods of Clem et al. The X (stellar-population) criterion of Schuster et al., which combines both kinematic and metallicity information, provides 22 thick-disk stars. \sigma_{\rm W} = 32 \pm 5 km s^{-1}, = 154 \pm 6 km s^{-1} and = -0.55 \pm 0.03 dex for these thick-disk stars, which is in agreement with values from previous studies of the thick disk. \alpha -element abundances which are available for some of these thick-disk stars show the typical alpha-element signatures of the thick disk, supporting the classification procedure based on the $X$ criteria. Both the scatter in metallicity at a given age and the presence of old, metal-rich stars in the age-metallicity relation make it difficult to decide whether or not an age-metallicity relation exists for the older thin-disk stars. For ages greater than 3 Gyr, our results agree with the other recent studies that there is almost no correlation between age and metallicity, \Delta ([M/Fe])/\Delta(age) = -0.01 \pm 0.005 dex Gyr^{-1}. For the 22 thick-disk stars there is a range in ages of 7-8 Gyr, but again almost no correlation between age and metallicity.Comment: 11 pages, including 10 figures and 3 tables, accepted for publication in MNRA

Theory for Metal Hydrides with Switchable Optical Properties

Recently it has been discovered that lanthanum, yttrium, and other metal hydride films show dramatic changes in the optical properties at the metal-insulator transition. Such changes on a high energy scale suggest the electronic structure is best described by a local model based on negatively charged hydrogen (H$^-$) ions. We develop a many-body theory for the strong correlation in a H$^-$ ion lattice. The metal hydride is described by a large $U$-limit of an Anderson lattice model. We use lanthanum hydride as a prototype of these compounds, and find LaH$_3$ is an insulator with a substantial gap consistent with experiments. It may be viewed either as a Kondo insulator or a band insulator due to strong electron correlation. A H vacancy state in LaH$_3$ is found to be highly localized due to the strong bonding between the electron orbitals of hydrogen and metal atoms. Unlike the impurity states in the usual semiconductors, there is only weak internal optical transitions within the vacancy. The metal-insulator transition takes place in a band of these vacancy states.Comment: 18 pages, 16 figures and 6 tables. Submitted to PR