Role of G protein coupled inwardly rectifier K+ channels (GIRK) on the neurobiology depression

353 págs.

On the origin of shear banding instability in metallic glasses

To uncover the physical origin of shear-banding instability in metallic glass (MG), a theoretical description of thermo-mechanical deformation of MG undergoing one-dimensional simple shearing is presented. The coupled thermo-mechanical model takes into account the momentum balance, the energy balance and the dynamics of free volume. The interplay between free-volume production and temperature increase being two potential causes for shear-banding instability is examined on the basis of the homogeneous solution. It is found that the free-volume production facilitates the sudden increase in the temperature before instability and vice versa. A rigorous linear perturbation analysis is used to examine the inhomogeneous deformation, during which the onset criteria and the internal length and time scales for three types of instabilities, namely free-volume softening, thermal softening and coupling softening, are clearly revealed. The shear-banding instability originating from sole free-volume softening takes place easier and faster than that due to sole thermal softening, and dominates in the coupling softening. Furthermore, the coupled thermo-mechanical shear-band analysis does show that an initial slight distribution of local free volume can incur significant strain localization, producing a shear band. During such a localization process, the local free-volume creation occurs indeed prior to the increase in local temperature, indicating that the former is the cause of shear localization, whereas the latter is its consequence. Finally, extension of the above model to include the shear-induced dilatation shows that such dilatation facilitates the shear instability in metallic glasses.

Growth, movement, and attrition of northern bluefin tuna, Thunnus thynnus, in the Pacific Ocean, as determined by tagging

ENGLISH: The growth of northern bluefin tuna is described by a two-stanza model. For fish between 191 and 564 mm in length the Gompertz curve, with values of 581 mm and 4.32 for Loo and K (annual), respectively, is used. The fish between 564 and 1530 mm grow linearly, at the rate of 0.709 mm per day. Age-O fish tagged and released in the western Pacific Ocean have been recaptured in the western, central, and eastern Pacific. The minimum time between release in the western Pacific and recapture in the eastern Pacific is 215 days. Older fish, mostly Land 2-year olds, tagged and released in the eastern Pacific have been recaptured in the eastern and western Pacific. The minimum time between release in eastern Pacific and recapture in the western Pacific is 674 days. The coefficient of natural mortality is estimated from data on growth and ambient temperature to be 0.276 on an annual basis, with 90-percent confidence limits of 0.161 and 0.47L Spawning of northern bluefin takes place only in the western Pacific. Some of the juveniles migrate to the eastern Pacific, where they reside for several months to several years before returning to the western Pacific. The portion of fish which migrate to the eastern Pacific varies among years, and this appears to be an important cause of the annual variation in the catches in the eastern Pacific Ocean. SPANISH: El crecimiento del atún aleta azul del norte es descrito por un modelo de dos estadios. Para los peces de entre 191 y 564 mm de talla se usa la curva de Gompertz, con valores de 581 mm y 4.32 para Loo y K (anual), respectivamente. Los peces de entre 564 y 1530 mm crecen de forma lineal, a 0.709 mm por día. Peces de edad Omarcados y liberados en el Pacífico occidental han sido recapturados en el Pacífico occidental, central, y oriental. La demora mínima entre la liberación en el Pacífico occidental y la recaptura en el Pacífico oriental es de 215 días. Peces mayores, principalmente de 1 ó 2 años de edad, marcados y liberados en el Pacífico oriental han sido re capturados en el Pacífico occidental y oriental. La demora mínima entre la liberación en el Pacífico oriental y la recaptura en el Pacífico occidental es de 674 días. Se estima el coeficiente de mortalidad natural a partir de los datos de crecimiento y temperatura ambiental en un 0.276 anual, con límites de confianza al 90% de 0.161 y 0.471. El aleta azul del norte desova únicamente en el Pacífico occidental. Algunos de los juveniles migran al Pacífico oriental, donde permanecen entre varios meses y varios años antes de regresar al Pacífico occidental. La porción de los peces que migran al Pacífico oriental varía entre años, y ésto parece ser una causa importante de la variación anual en las capturas en el Océano Pacífico oriental. (PDF contains 94 pages.)

Direct observation of adsorption sites of protein impurities and their effects on step advancement of protein crystals

We measured noninvasively step velocities of elementary two-dimensional (2D) islands on {110} faces of tetragonal lysozyme crystals, under various supersaturations, by laser confocal microscopy combined with differential interference contrast microscopy. We studied the correlation between the effects of protein impurities on the growth of elementary steps and their adsorption sites on a crystal surface, using three kinds of proteins: fluorescent-labeled lysozyme (F-lysozyme), covalently bonded dimers of lysozyme (dimer), and a 18 kDa polypeptide (18 kDa). These three protein impurities suppressed the advancement of the steps. However, they exhibited different supersaturation dependencies of the suppression of the step velocities. To clarify the cause of this difference, we observed in situ the adsorption sites of individual molecules of F-lysozyme and fluorescent-labeled dimer (F-dimer) on the crystal surface by single-molecule visualization. We found that F-lysozyme adsorbed preferentially on steps (i.e., kinks), whereas F-dimer adsorbed randomly on terraces. Taking into account the different adsorption sites of F-lysozyme and F-dimer, we could successfully explain the different effects of the impurities on the step velocities. These observations strongly suggest that 18 kDa also adsorbs randomly on terraces. Seikagaku lysozyme exhibited a complex effect that could not alone be explained by the two major impurities (dimer and 18 kDa) present in Seikagaku lysozyme, indicating that trace amounts of other impurities significantly affect the step advancement.

Stormwater runoff - modeling impacts of urbanization and climate change

Development pressure throughout the coastal areas of the United States continues to build, particularly in the southeast (Allen and Lu 2003, Crossett et al. 2004). It is well known that development alters watershed hydrology: as land becomes covered with surfaces impervious to rain, water is redirected from groundwater recharge and evapotranspiration to stormwater runoff, and as the area of impervious cover increases, so does the volume and rate of runoff (Schueler 1994, Corbett et al. 1997). Pollutants accumulate on impervious surfaces, and the increased runoff with urbanization is a leading cause of nonpoint source pollution (USEPA 2002). Sediment, chemicals, bacteria, viruses, and other pollutants are carried into receiving water bodies, resulting in degraded water quality (Holland et al. 2004, Sanger et al. 2008). (PDF contains 5 pages)

Wind-driven desertification: process modeling, remote monitoring, and forecasting

Arid and semiarid landscapes comprise nearly a third of the Earth's total land surface. These areas are coming under increasing land use pressures. Despite their low productivity these lands are not barren. Rather, they consist of fragile ecosystems vulnerable to anthropogenic disturbance.

The purpose of this thesis is threefold: (I) to develop and test a process model of wind-driven desertification, (II) to evaluate next-generation process-relevant remote monitoring strategies for use in arid and semiarid regions, and (III) to identify elements for effective management of the world's drylands.

In developing the process model of wind-driven desertification in arid and semiarid lands, field, remote sensing, and modeling observations from a degraded Mojave Desert shrubland are used. This model focuses on aeolian removal and transport of dust, sand, and litter as the primary mechanisms of degradation: killing plants by burial and abrasion, interrupting natural processes of nutrient accumulation, and allowing the loss of soil resources by abiotic transport. This model is tested in field sampling experiments at two sites and is extended by Fourier Transform and geostatistical analysis of high-resolution imagery from one site.

Next, the use of hyperspectral remote sensing data is evaluated as a substantive input to dryland remote monitoring strategies. In particular, the efficacy of spectral mixture analysis (SMA) in discriminating vegetation and soil types and detennining vegetation cover is investigated. The results indicate that hyperspectral data may be less useful than often thought in determining vegetation parameters. Its usefulness in determining soil parameters, however, may be leveraged by developing simple multispectral classification tools that can be used to monitor desertification.

Finally, the elements required for effective monitoring and management of arid and semiarid lands are discussed. Several large-scale multi-site field experiments are proposed to clarify the role of wind as a landscape and degradation process in dry lands. The role of remote sensing in monitoring the world's drylands is discussed in terms of optimal remote sensing platform characteristics and surface phenomena which may be monitored in order to identify areas at risk of desertification. A desertification indicator is proposed that unifies consideration of environmental and human variables.

Functional evaluation of noncovalent interactions in neuroreceptors and progress toward the expansion of unnatural amino acid methodology

This dissertation primarily describes chemical-scale studies of G protein-coupled receptors and Cys-loop ligand-gated ion channels to better understand ligand binding interactions and the mechanism of channel activation using recently published crystal structures as a guide. These studies employ the use of unnatural amino acid mutagenesis and electrophysiology to measure subtle changes in receptor function.

In chapter 2, the role of a conserved aromatic microdomain predicted in the D3 dopamine receptor is probed in the closely related D2 and D4 dopamine receptors. This domain was found to act as a structural unit near the ligand binding site that is important for receptor function. The domain consists of several functionally important noncovalent interactions including hydrogen bond, aromatic-aromatic, and sulfur-π interactions that show strong couplings by mutant cycle analysis. We also assign an alternate interpretation for the linear fluorination plot observed at W6.48, a residue previously thought to participate in a cation-π interaction with dopamine.

Chapter 3 outlines attempts to incorporate chemically synthesized and in vitro acylated unnatural amino acids into mammalian cells. While our attempts were not successful, method optimizations and data for nonsense suppression with an in vivo acylated tRNA are included. This chapter is aimed to aid future researchers attempting unnatural amino acid mutagenesis in mammalian cells.

Chapter 4 identifies a cation-π interaction between glutamate and a tyrosine residue on loop C in the GluClβ receptor. Using the recently published crystal structure of the homologous GluClα receptor, other ligand-binding and protein-protein interactions are probed to determine the similarity between this invertebrate receptor and other more distantly related vertebrate Cys-loop receptors. We find that many of the interactions previously observed are conserved in the GluCl receptors, however care must be taken when extrapolating structural data.

Chapter 5 examines inherent properties of the GluClα receptor that are responsible for the observed glutamate insensitivity of the receptor. Chimera synthesis and mutagenesis reveal the C-terminal portion of the M4 helix and the C-terminus as contributing to formation of the decoupled state, where ligand binding is incapable of triggering channel gating. Receptor mutagenesis was unable to identify single residue mismatches or impaired protein-protein interactions within this domain. We conclude that M4 helix structure and/or membrane dynamics are likely the cause of ligand insensitivity in this receptor and that the M4 helix has an role important in the activation process.

Fundamental Studies of Carbon Oxygen Bond Activation in Nickel Diphosphine Ether Complexes. And, Metallomacrocycles as Ligands: Synthesis and Characterization of Aluminum-Bridged Bisglyoximato Complexes of Iron and Cobalt

In order to develop better catalysts for the cleavage of aryl-X bonds fundamental studies of the mechanism and individual steps of the mechanism have been investigated in detail. As the described studies are difficult at best in catalytic systems, model systems are frequently used. To study aryl-oxygen bond activation, a terphenyl diphosphine scaffold containing an ether moiety in the central arene was designed. The first three chapters of this dissertation focus on the studies of the nickel complexes supported by this diphosphine backbone and the research efforts in regards to aryl-oxygen bond activation.

Chapter 2 outlines the synthesis of a variety of diphosphine terphenyl ether ligand scaffolds. The metallation of these scaffolds with nickel is described. The reactivity of these nickel(0) systems is also outlined. The systems were found to typically undergo a reductive cleavage of the aryl oxygen bond. The mechanism was found to be a subsequent oxidative addition, β-H elimination, reductive elimination and (or) decarbonylation.

Chapter 3 presents kinetic studies of the aryl oxygen bond in the systems outlined in Chapter 2. Using a series of nickel(0) diphosphine terphenyl ether complexes the kinetics of aryl oxygen bond activation was studied. The activation parameters of oxidative addition for the model systems were determined. Little variation was observed in the rate and activation parameters of oxidative addition with varying electronics in the model system. The cause of the lack of variation is due to the ground state and oxidative addition transition state being affected similarly. Attempts were made to extend this study to catalytic systems.

Chapter 4 investigates aryl oxygen bond activation in the presence of additives. It was found that the addition of certain metal alkyls to the nickel(0) model system lead to an increase in the rate of aryl oxygen bond activation. The addition of excess Grignard reagent led to an order of magnitude increase in the rate of aryl oxygen bond activation. Similarly the addition of AlMe3 led to a three order of magnitude rate increase. Addition of AlMe₃ at -80 °C led to the formation of an intermediate which was identified by NOESY correlations as a system in which the AlMe3 is coordinated to the ether moiety of the backbone. The rates and activation parameters of aryl oxygen bond activation in the presence of AlMe3 were investigated.

The last two chapters involve the study of metalla-macrocycles as ligands. Chapter 5 details the synthesis of a variety of glyoxime backbones and diphenol precursors and their metallation with aluminum. The coordination chemistry of iron on the aluminum scaffolds was investigated. Varying the electronics of the aluminum macrocycle was found to affect the observed electrochemistry of the iron center.

Chapter 6 extends the studies of chapter 5 to cobalt complexes. The synthesis of cobalt dialuminum glyoxime metal complexes is described. The electrochemistry of the cobalt complexes was investigated. The electrochemistry was compared to the observed electrochemistry of a zinc analog to identify the redox activity of the ligand. In the presence of acid the cobalt complexes were found to electrochemically reduce protons to dihydrogen. The electronics of the ancillary aluminum ligands were found to affect the potential of proton reduction in the cobalt complexes. These potentials were compared to other diglyoximate complexes.

The isotopic composition of energetic particles emitted from a large solar flare

We report measurements of isotope abundance ratios for 5-50 MeV/nuc nuclei from a large solar flare that occurred on September 23, 1978. The measurements were made by the Heavy Isotope Spectrometer Telescope (HIST) on the ISEE-3 satellite orbiting the Sun near an Earth-Sun libration point approximately one million miles sunward of the Earth. We report finite values for the isotope abundance ratios ¹³C/¹²C, ¹⁵N/¹⁴N, ¹⁸O/¹⁶O, ²²Ne/ ²⁰Ne, ²⁵Mg/²⁴Mg, and ²⁶Mg/²⁴Mg, and upper limits for the isotope abundance ratios ³He/⁴He, ¹⁴C/¹²C, ¹⁷O/¹⁶O, and ²¹Ne/²⁰Ne.

We measured element abundances and spectra to compare the September 23, 1978 flare with other flares reported in the literature. The flare is a typical large flare with "low" Fe/O abundance (≤ 0.1).

For ¹³C/¹²C, ¹⁵N/¹⁴N, ¹⁸O/¹⁶O, ²⁵Mg/ ²⁴Mg, and ²⁶Mg/²⁴Mg, our measured isotope abundance ratios agree with the solar system abundance ratios of Cameron (1981). For neon we measure ²²Ne/²⁰Ne = 0.109 + 0.026 - 0.019, a value that is different with confidence 97.5% from the abundance measured in the solar wind by Geiss at al. (1972) of ²²Ne/²⁰Ne = 0.073 ± 0.001. Our measurement for ²²Ne/²⁰Ne agrees with the isotopic composition of the meteoritic component neon-A.

Separate arguments appear to rule out simple mass fractionation in the solar wind and in our solar energetic particle measurements as the cause of the discrepancy in the comparison of the apparent compositions of these two sources of solar material.

Stationary Eddies and Zonal Variations of the Global Hydrological Cycle in a Changing Climate

This thesis advances our physical understanding of the sensitivity of the hydrological cycle to global warming. Specifically, it focuses on changes in the longitudinal (zonal) variation of precipitation minus evaporation (P - E), which is predominantly controlled by planetary-scale stationary eddies. By studying idealized general circulation model (GCM) experiments with zonally varying boundary conditions, this thesis examines the mechanisms controlling the strength of stationary-eddy circulations and their role in the hydrological cycle. The overarching goal of this research is to understand the cause of changes in regional P - E with global warming. An understanding of such changes can be useful for impact studies focusing on water availability, ecosystem management, and flood risk.

Based on a moisture-budget analysis of ERA-Interim data, we establish an approximation for zonally anomalous P - E in terms of surface moisture content and stationary-eddy vertical motion in the lower troposphere. Part of the success of this approximation comes from our finding that transient-eddy moisture fluxes partially cancel the effect of stationary-eddy moisture advection, allowing divergent circulations to dominate the moisture budget. The lower-tropospheric vertical motion is related to horizontal motion in stationary eddies by Sverdrup and Ekman balance. These moisture- and vorticity-budget balances also hold in idealized and comprehensive GCM simulations across a range of climates.

By examining climate changes in the idealized and comprehensive GCM simulations, we are able to show the utility of the vertical motion P - E approximation for splitting changes in zonally anomalous P - E into thermodynamic and dynamic components. Shifts in divergent stationary-eddy circulations dominate changes in zonally anomalous P - E. This limits the local utility of the "wet gets wetter, dry gets drier” idea, where existing P - E patterns are amplified with warming by the increase in atmospheric moisture content, with atmospheric circulations held fixed. The increase in atmospheric moisture content manifests instead in an increase in the amplitude of the zonally anomalous hydrological cycle as measured by the zonal variance of P - E. However, dynamic changes, particularly the slowdown of divergent stationary-eddy circulations, limit the strengthening of the zonally anomalous hydrological cycle. In certain idealized cases, dynamic changes are even strong enough to reverse the tendency towards "wet gets wetter, dry gets drier” with warming.

Motivated by the importance of stationary-eddy vertical velocities in the moisture budget analysis, we examine controls on the amplitude of stationary eddies across a wide range of climates in an idealized GCM with simple topographic and ocean-heating zonal asymmetries. An analysis of the thermodynamic equation in the vicinity of topographic forcing reveals the importance of on-slope surface winds, the midlatitude isentropic slope, and latent heating in setting the amplitude of stationary waves. The response of stationary eddies to climate change is determined primarily by the strength of zonal surface winds hitting the mountain. The sensitivity of stationary-eddies to this surface forcing increases with climate change as the slope of midlatitude isentropes decreases. However, latent heating also plays an important role in damping the stationary-eddy response, and this damping becomes stronger with warming as the atmospheric moisture content increases. We find that the response of tropical overturning circulations forced by ocean heat-flux convergence is described by changes in the vertical structure of moist static energy and deep convection. This is used to derive simple scalings for the Walker circulation strength that capture the monotonic decrease with warming found in our idealized simulations.

Through the work of this thesis, the advances made in understanding the amplitude of stationary-waves in a changing climate can be directly applied to better understand and predict changes in the zonally anomalous hydrological cycle.

A control of gas-vacuolate cyanobacteria

The cyanobacteria that cause problems in water supply are principally the colonial forms that are buoyed up by gas vesicles. The success of these organisms is due, in part, to their gas vesicles, which enable them to perform vertical migrations or to maintain themselves in the euphotic zone. The gas vesicles are also the root cause of the problems. In calm periods they cause the cyanobacteria to float to the water surface forming noxious scums, and they may prevent the colonies from sedimenting in water treatment plants. Gas vesicles are hollow, gas-filled structures; they are rigid but can be collapsed by the application of pressure. Their critical collapse pressure is influenced by their dimensions, which vary in different organisms. Gas vesicles are formed by the assembly of two types of protein, which determine their mechanical and physical properties. Methods for collapsing gas vesicles in natural populations of cyanobacteria will be considered. They may have application to the control of cyanobacteria in water supply.

Perinatal hipoxic ischemic injury in the auditory pathway and the effect of several neuroprotective agents

192 p.

Genotypes, Recombinant Forms, and Variants of Norovirus GII.4 in Gipuzkoa (Basque Country, Spain), 2009-2012

Background: Noroviruses (NoVs) are genetically diverse, with genogroup II-and within it-genotype 4 (GII.4) being the most prevalent cause of acute gastroenteritis worldwide. The aim of this study was to characterize genogroup II NoV causing acute gastroenteritis in the Basque Country (northern Spain) from 2009-2012. Methods: The presence of NoV RNA was investigated by reverse transcriptase-polymerase chain reaction (RT-PCR) in stool specimens from children younger than 15 years old with community-acquired acute gastroenteritis, and from hospitalized adults or elderly residents of nursing homes with acute gastroenteritis. For genotyping, the open reading frames ORF1 (encoding the polymerase) and ORF2 (encoding the major capsid protein) were partially amplified and sequenced. Recombinant strains were confirmed by PCR of the ORF1/ORF2 junction region. Results: NoV was detected in 16.0% (453/2826) of acute gastroenteritis episodes in children younger than 2 years, 9.9% (139/1407) in children from 2 to 14 years, and 35.8% (122/341) in adults. Of 317 NoVs characterized, 313 were genogroup II and four were genogroup I. The GII.4 variants Den Haag-2006b and New Orleans-2009 predominated in 2009 and 2010-2011, respectively. In 2012, the New Orleans-2009 variant was partially replaced by the Sydney-2012 variant (GII.Pe/GII.4) and New Orleans-2009/Sydney-2012 recombinant strains. The predominant capsid genotype in all age groups was GII.4, which was the only genotype detected in outbreaks. The second most frequent genotype was GII.3 (including the recently described recombination GII.P16/GII.3), which was detected almost exclusively in children. Conclusion: Nine different genotypes of NoV genogroup II were detected; among these, intergenotype recombinant strains represented an important part, highlighting the role of recombination in the evolution of NoVs. Detection of new NoV strains, not only GII.4 strains, shortly after their first detection in other parts of the world shows that many NoV strains can spread rapidly.

Carbapenem-resistant and carbapenem-susceptible isogenic isolates of Klebsiella pneumoniae ST101 causing infection in a tertiary hospital

Background: In this study we describe the clinical and molecular characteristics of an outbreak due to carbapenem-resistant Klebsiella pneumoniae (CR-KP) producing CTX-M-15 and OXA-48 carbapenemase. Isogenic strains, carbapenem-susceptible K. pneumoniae (CS-KP) producing CTX-M-15, were also involved in the outbreak. Results: From October 2010 to December 2012 a total of 62 CR-KP and 23 CS-KP were isolated from clinical samples of 42 patients (22 had resistant isolates, 14 had susceptible isolates, and 6 had both CR and CS isolates). All patients had underlying diseases and 17 of them (14 patients with CR-KP and 3 with CS-KP) had received carbapenems previously. The range of carbapenem MICs for total isolates were: imipenem: 2 to >32 mu g/ml vs. <2 mu g/ml; meropenem: 4 to >32 mu g/ml vs. <2 mu g/ml; and ertapenem: 8 to >32 mu g/ml vs. <2 mu g/ml. All the isolates were also resistant to gentamicin, ciprofloxacin, and cotrimoxazole. Both types of isolates shared a common PFGE pattern associated with the multilocus sequence type 101 (ST101). The bla(CTX-M-15) gene was detected in all the isolates, whereas the bla(OXA-48) gene was only detected in CR-KP isolates on a 70 kb plasmid. Conclusions: The clonal spread of K. pneumoniae ST101 expressing the OXA-48 and CTX-M-15 beta-lactamases was the cause of an outbreak of CR-KP infections. CTX-M-15-producing isolates lacking the blaOXA-48 gene coexisted during the outbreak.

Temporal variation in growth of yellowfin tuna (Thunnus albacares) larvae in the Panama Bight, 1990

Tuna larvae (at flexion, postflexion, and transformation stages) were collected by dip net and light traps at night in the northwestern Panama Bight during the season of reduced upwelling (June−September) of 1990, 1991, 1992, and 1997. The larvae were identified as yellowfin tuna (Thunnus albacares) by mtDNA analysis. Ichthyoplankton data from bongo and Tucker trawl tows were used to examine the potential prey abundance in relation to the mean size-at-age and growth rates of the yellowfin tuna larvae and their otoliths. The most rapid growth rates occurred during June 1990 when plankton volumes were at their highest levels. The lowest plankton volumes coincided with the lowest growth rates and mean sizes-at-age during the August−September 1991 period. High densities of larval fish were prevalent in the ichthyoplankton tows during the 1991 period; therefore intra- and interspecific competition for limited food resources may have been the cause of slower growth (density-dependent growth) in yellowfin tuna larvae The highest mean seasurface temperature and the lowest mean wind stress occurred during an El Niño-Southern Oscillation (ENSO) event during the 1997 period. There appeared to be no clear association between these environmental factors and larval growth rates, but the higher temperatures may have caused an increase in the short-term growth of otoliths in relat