Nuclease-based editing in the porcine genome : a strategy to facilitate porcine-to human xenotransplantation

Indiana University-Purdue University Indianapolis (IUPUI)Solid organ transplantation is severely limited by a shortage of available donor allografts. Pig-to-human xenotransplantation offers a potential solution to this growing problem. For xenotransplantation to achieve clinical relevance, both immunologic and physiologic barriers must be understood. Genetic modification of pigs has proven to be a valuable means of both studying and eliminating these barriers. The present body of work describes a method for greatly increasing the efficiency and precision of genome editing within the porcine genome. By combining non-integrating selection and homologous recombination of exogenous oligonucleotides, a method for rapidly creating genetic modification without reliance on phenotypic sorting was achieved. Furthermore this work employs the technique of CRISPR/Cas9-directed mutagenesis to create and analyze several new animal models of porcine-to-human xenotransplantation with respect to both immunologic and physiologic parameters. First, Isoglobotrihexosylceramide -a controversial glycan to the field of xenotransplantation- was studied in a knockout model and found not to affect human-anti-porcine humoral reactions. Second, a new combination of glycan modifications is described that significantly lowers the human anti-porcine humoral immune response. This model animal suggests that glycan-deletion alone will be sufficient to promote clinical application, and that conventional immunosuppression will be successful in mediating the human cellular response. Finally, two potential physiologic barriers to xenotransplantation are studied in genetically modified model animals. Xenogenic consumption of human platelets was studied across hepatic and renal organ systems; xenogenic platelet consumption was reduced by glycan modifications to the porcine liver while human platelet sequestration was not identified in the study of renal endothelium. Porcine FcRN –an essential receptor expressed in kidneys to maintain serum proteostasis- was studied as a final potential barrier to pig-to human renal transplantation. Because albumin is the primary driver of serum oncotic pressure, the protein-protein interaction of endogenous porcine FcRN and human albumin was studied. Porcine FcRN was found capable of binding human albumin under physiologic parameters. In summary, the results of the present work suggest that the salient barriers to clinical xenotransplantation have been removed and that porcine-to human renal transplantation may soon offer an answer to the current organ shortage

Deriving Iodine-free spectra for high-resolution echelle spectrographs

We describe a new method to derive clean, iodine-free spectra directly from observations acquired using high-resolution echelle spectrographs equipped with iodine cells. The main motivation to obtain iodine-free spectra is to use portions of the spectrum that are superimposed with the dense forest of iodine absorption lines, in order to retrieve lines that can be used to monitor the magnetic activity of the star, helping to validate candidate planets. In short, we provide a straight-forward methodology to clean the spectra by using the forward model used to derive radial velocities, the Line Spread Function information plus the stellar spectrum without iodine to reconstruct and subtract the iodine spectrum from the observations. We show our results using observations of the star $\tau$ Ceti acquired with the PFS, HIRES and UCLES spectrographs, reaching an iodine-free spectrum correction at the $\sim$1% RMS level. We additionally discuss the limitations and further applications of the method.Comment: 15 pages, 7 figures. Accepted for publication in A

Quenched Chiral Perturbation Theory for Baryons

We develop quenched chiral perturbation theory for baryons using the graded-symmetry formalism of Bernard and Golterman and calculate non-analytic contributions to the baryon masses coming from quenched chiral loops. The usual term proportional to $m_{q}^{3/2}$ is substantially altered due to the cancellation of diagrams with internal quark loops. In addition, the $\eta'$ ``hairpin'' vertex leads to a new correction, proportional to $m_{q}^{1/2}$. We compare our results to numerical lattice data and use them to estimate the size of the quenching error in the octet baryon masses.Comment: 7 pages (An abridged version of this note will appear in the proceedings of Lattice'93. Latex + 14 postscript files, bundled using uufiles. Needs psfig.) UW/PT-93-0

Less government intervention in biodiversity management: risks and opportunities

n a changing global environment, with increasing pressure on ecosystem goods and services, biodiversity conservation is likely to become increasingly important. However, with the current global financial crisis, governments are increasingly trying to stabilise economies through spending cuts aiming to reduce national deficits. Within such an economic climate, the devolution of governance through public participation is an intrinsically appealing concept. We outline a number of challenges that explain why increased participation in biodiversity management has been and may continue to be problematic. Using as a case study the local stakeholder-driven Moray Firth Seal Management Plan in Scotland, we identify four key conditions that were crucial to the successful participatory management of a biodiversity conflict: a local champion, the emergence of a crisis point, the involvement of decision-makers, and long-term financial and institutional support. Three of the four conditions point to the role of direct government involvement, highlighting the risk of devolving responsibility for biodiversity conflict management to local communities. We argue that without an informed debate, the move towards a more participatory approach could pose a danger to hard-won policy gains in relation to public participation, biodiversity conservation and conflict management

Crossing the Brown Dwarf Desert Using Adaptive Optics: A Very Close L-Dwarf Companion to the Nearby Solar Analog HR 7672

We have found a very faint companion to the active solar analog HR 7672 (HD 190406; GJ 779; 15 Sge). Three epochs of high resolution imaging using adaptive optics (AO) at the Gemini-North and Keck II Telescopes demonstrate that HR 7672B is a common proper motion companion, with a separation of 0.79" (14 AU) and a 2.16 um flux ratio of 8.6 mags. Using follow-up K-band spectroscopy from Keck AO+NIRSPEC, we measure a spectral type of L4.5+/-1.5. This is the closest ultracool companion around a main sequence star found to date by direct imaging. We estimate the primary has an age of 1-3 Gyr. Assuming coevality, the companion is most likely substellar, with a mass of 55-78 Mjup based on theoretical models. The primary star shows a long-term radial velocity trend, and we combine the radial velocity data and AO imaging to set a firm (model-independent) lower limit of 48 Mjup. In contrast to the paucity of brown dwarf companions at <~4 AU around FGK dwarfs, HR 7672B implies that brown dwarf companions do exist at separations comparable to those of the giant planets in our own solar system. Its presence is at variance with scenarios where brown dwarfs form as ejected stellar embryos. Moreover, since HR 7672B is likely too massive to have formed in a circumstellar disk as planets are believed to, its discovery suggests that a diversity of physical processes act to populate the outer regions of exoplanetary systems.Comment: Astrophysical Journal, in pres

Aerobrake assembly with minimum Space Station accommodation

The minimum Space Station Freedom accommodations required for initial assembly, repair, and refurbishment of the Lunar aerobrake were investigated. Baseline Space Station Freedom support services were assumed, as well as reasonable earth-to-orbit possibilities. A set of three aerobrake configurations representative of the major themes in aerobraking were developed. Structural assembly concepts, along with on-orbit assembly and refurbishment scenarios were created. The scenarios were exercised to identify required Space Station Freedom accommodations. Finally, important areas for follow-on study were also identified

Metal-ligand interplay in strongly-correlated oxides: a parametrized phase diagram for pressure induced spin transitions

We investigate the magnetic properties of archetypal transition-metal oxides MnO, FeO, CoO and NiO under very high pressure by x-ray emission spectroscopy at the K\beta line. We observe a strong modification of the magnetism in the megabar range in all the samples except NiO. The results are analyzed within a multiplet approach including charge-transfer effects. The pressure dependence of the emission line is well accounted for by changes of the ligand field acting on the d electrons and allows us to extract parameters like local d-hybridization strength, O-2p bandwidth and ionic crystal field across the magnetic transition. This approach allows a first-hand insight into the mechanism of the pressure induced spin transition.Comment: 5 pages, 3 figure

The Long Road to Developing Native Herbaceous Summer Forage Legume Ecotypes

Only a handful of well-adapted herbaceous summer forage legumes are currently marketed for drier regions of North America and even fewer are true natives. There is a growing demand for native germplasm in the region as a new generation of landowner attempts to return grasslands to a semblance of their original species and diversity. The objective of this paper is to describe preliminary research results of a grasslands team collecting, studying and promulgating native leguminous germplasm in Texas

Inequalities in bariatric surgery in Australia: findings from 49,364 obese participants in a prospective cohort study

OBJECTIVES: To investigate variation, and quantify socioeconomic inequalities, in receipt of primary bariatric surgery in an obese population. DESIGN, SETTING AND PARTICIPANTS: Prospective population-based cohort study of 49,364 individuals aged 45-74 with body mass index (BMI) ≥30kg/m2, with questionnaire (2006-09) linked to hospital and death data to July 2010. Sample drawn from the 45 and Up Study (~10% of NSW population aged ≥45 included, response rate ~18%). MAIN OUTCOME MEASURES: Rates of bariatric surgery and adjusted rate ratios (RR) in relation to health and sociodemographic characteristics. RESULTS: Over 111,757 person-years (py) of follow-up, 312 participants underwent bariatric surgery, a rate of 27.92/10,000 py (95%CI: 24.91-31.19). Rates were highest in women, people living in major cities and those with diabetes, and increased significantly with increasing body-mass-index and number of chronic health conditions. Adjusted RRs were: 5.27 (3.18-8.73) for annual household-income ≥$70,000 versus <$20,000; and 4.01 (2.41-6.67) for those living in areas in the least- versus most-disadvantaged quintile. Private health insurance (PHI) coverage (age-sex adjusted RR: 9.25; 5.70-15.00) partially explained the observed socioeconomic inequalities. CONCLUSIONS: Bariatric surgery has been shown to be cost-effective in treating severe obesity and associated illnesses. While bariatric surgery rates in Australia are higher in those with health problems, large socioeconomic inequalities are apparent. Our findings suggest these procedures are largely available to those who can afford PHI and associated out-of-pocket costs, with poor access in populations who are most in need. Continuing inequalities in access are likely to exacerbate existing inequalities in obesity and related health problems.National Health and Medical Research Council (NHMRC

Dynamic interpretation of slug tests in highly permeable aquifers

This is the published version. Copyright American Geophysical Union[1] Considerable progress has been made in developing a theoretical framework for modeling slug test responses in formations with high hydraulic conductivity K. However, several questions of practical significance remain unresolved. Given the rapid and often oscillatory nature of test responses, the traditional hydrostatic relationship between the water level and the transducer-measured head in the water column may not be appropriate. A general dynamic interpretation is proposed that describes the relationship between water level response and transducer-measured head. This theory is utilized to develop a procedure for transforming model-generated water level responses to transducer readings. The magnitude of the difference between the actual water level position and the apparent position based on the transducer measurement is a function of the acceleration and velocity of the water column, test geometry, and depth of the transducer. The dynamic approach explains the entire slug test response, including the often-noted discrepancy between the actual initial water level displacement and that measured by a transducer in the water column. Failure to use this approach can lead to a significant underestimation of K when the transducer is a considerable distance below the static water level. Previous investigators have noted a dependence of test responses on the magnitude of the initial water level displacement and have developed various approximate methods for analyzing such data. These methods are re-examined and their limitations clarified. Practical field guidelines are proposed on the basis of findings of this work. The soundness of the dynamic approach is demonstrated through a comparison of K profiles from a series of multilevel slug tests with those from dipole-flow tests performed in the same wells