Pharmacology and pore-forming domains of the cystic fibrosis transmembrane conductance regulator

The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel member of the ATP-binding cassette (ABC) superfamily of membrane proteins. CFTR has two homologous halves, each consisting of six transmembrane spanning domains (TM) followed by a nucleotide binding fold, connected by a regulatory (R) domain. This thesis addresses the question of which domains are responsible for Cl^- selectivity, i.e., which domains line the channel pore.

To address this question, novel blockers of CFTR were characterized. CFTR was heterologously expressed in Xenopus oocytes to study the mechanism of block by two closely related arylaminobenzoates, diphenylamine-2-carboxylic acid (DPC) and flufenamic acid (FFA). Block by both is voltage-dependent, with a binding site ≈ 40% through the electric field of the membrane. DPC and FFA can both reach their binding site from either side of the membrane to produce a flickering block of CFTR single channels. In addition, DPC block is influenced by Cl^- concentration, and DPC blocks with a bimolecular forward binding rate and a unimolecular dissociation rate. Therefore, DPC and FFA are open-channel blockers of CFTR, and a residue of CFTR whose mutation affects their binding must line the pore.

Screening of site-directed mutants for altered DPC binding affinity reveals that TM-6 and TM-12 line the pore. Mutation of residue 5341 in TM-6 abolishes most DPC block, greatly reduces single-channel conductance, and alters the direction of current rectification. Additional residues are found in TM-6 (K335) and TM-12 (T1134) whose mutations weaken or strengthen DPC block; other mutations move the DPC binding site from TM-6 to TM-12. The strengthened block and lower conductance due to mutation T1134F is quantitated at the single-channel level. The geometry of DPC and of the residues mutated suggest α-helical structures for TM-6 and TM-12. Evidence is presented that the effects of the mutations are due to direct side-chain interaction, and not to allosteric effects propagated through the protein. Mutations are also made in TM-11, including mutation S1118F, which gives voltage-dependent current relaxations. The results may guide future studies on permeation through ABC transporters and through other Cl^- channels.

Study of the origins of toughness in amorphous metals

Amorphous metals that form fully glassy parts over a few millimeters in thickness are still relatively new materials. Their glassy structure gives them particularly high strengths, high yield strains, high hardness values, high resilience, and low damping losses, but this can also result in an extremely low tolerance to the presence of flaws in the material. Since this glassy structure lacks the ordered crystal structure, it also lacks the crystalline defect (dislocations) that provides the micromechanism of toughening and flaw insensitivity in conventional metals. Without a sufficient and reliable toughness that results in a large tolerance of damage in the material, metallic glasses will struggle to be adopted commercially. Here, we identify the origin of toughness in metallic glass as the competition between the intrinsic toughening mechanism of shear banding ahead of a crack and crack propagation by the cavitation of the liquid inside the shear bands. We present a detailed study over the first three chapters mainly focusing on the process of shear banding; its crucial role in giving rise to one of the most damage-tolerant materials known, its extreme sensitivity to the configurational state of a glass with moderate toughness, and how the configurational state can be changed with the addition of minor elements. The last chapter is a novel investigation into the cavitation barrier in glass-forming liquids, the competing process to shear banding. The combination of our results represents an increased understanding of the major influences on the fracture toughness of metallic glasses and thus provides a path for the improvement and development of tougher metallic glasses.

I. Geology of the Southwest quarter of the Elsinore Quadrangle. II. Geochemical properties of the waters of the Elsinore Quadrangle [= Geology of the Lake Elsinore Quadrangle, California. Mineral deposits of Lake Elsinore Quadrangle, California]

The Lake Elsinore quadrangle covers about 250 square miles and includes parts of the southwest margin of the Perris Block, the Elsinore trough, the southeastern end of the Santa Ana Mountains, and the Elsinore Mountains.

The oldest rocks consist of an assemblage of metamorphics of igneous effusive and sedimentary origin, probably, for the most part, of Triassic age. They are intruded by diorite and various hypabyssal rocks, then in turn by granitic rocks, which occupy over 40 percent of the area. Following this last igneous activity of probable Lower Cretaceous age, an extended period of sedimentation started with the deposition of the marine Upper Cretaceous Chico formation and continued during the Paloecene under alternating marine and continental conditions on the margins of the blocks. A marine regression towards the north, during the Neocene, accounts for the younger Tertiary strata in the region under consideration.

Outpouring of basalts to the southeast indicates that igneous activity was resumed toward the close of the Tertiary. The fault zone, which characterizes the Elsinor trough, marks one of the major tectonic lines of southem California. It separates the upthrown and tilted block of the Santa Ana Mountains to the south from the Perris Block to the north.

Most of the faults are normal in type and nearly parallel to the general trend of the trough, or intersect each other at an acute angle. Vertical displacements generally exceed the horizontal ones and several periods of activity are recognized.

Tilting of Tertiary and older Quaternary sediments in the trough have produced broad synclinal structures which have been modified by subsequent faulting.

Five old surfaces of erosion are exposed on the highlands.

The mineral resources of the region are mainly high-grade clay deposits and mineral waters.

Characterization of the human SCF ubiquitin ligases - structure, function, and regulation

The SCF ubiquitin ligase complex of budding yeast triggers DNA replication by cata lyzi ng ubiquitination of the S phase CDK inhibitor SIC1. SCF is composed of several evolutionarily conserved proteins, including ySKP1, CDC53 (Cullin), and the F-box protein CDC4. We isolated hSKP1 in a two-hybrid screen with hCUL1, the human homologue of CDC53. We showed that hCUL1 associates with hSKP1 in vivo and directly interacts with hSKP1 and the human F-box protein SKP2 in vitro, forming an SCF-Iike particle. Moreover, hCUL1 complements the growth defect of yeast CDC53^(ts) mutants, associates with ubiquitination-promoting activity in human cell extracts, and can assemble into functional, chimeric ubiquitin ligase complexes with yeast SCF components. These data demonstrated that hCUL1 functions as part of an SCF ubiquitin ligase complex in human cells. However, purified human SCF complexes consisting of CUL1, SKP1, and SKP2 are inactive in vitro, suggesting that additional factors are required.

Subsequently, mammalian SCF ubiquitin ligases were shown to regulate various physiological processes by targeting important cellular regulators, like lĸBα, β-catenin, and p27, for ubiquitin-dependent proteolysis by the 26S proteasome. Little, however, is known about the regulation of various SCF complexes. By using sequential immunoaffinity purification and mass spectrometry, we identified proteins that interact with human SCF components SKP2 and CUL1 in vivo. Among them we identified two additional SCF subunits: HRT1, present in all SCF complexes, and CKS1, that binds to SKP2 and is likely to be a subunit of SCF5^(SKP2) complexes. Subsequent work by others demonstrated that these proteins are essential for SCF activity. We also discovered that COP9 Signalosome (CSN), previously described in plants as a suppressor of photomorphogenesis, associates with CUL1 and other SCF subunits in vivo. This interaction is evolutionarily conserved and is also observed with other Cullins, suggesting that all Cullin based ubiquitin ligases are regulated by CSN. CSN regulates Cullin Neddylation presumably through CSNS/JAB1, a stochiometric Signalosome subunit and a putative deneddylating enzyme. This work sheds light onto an intricate connection that exists between signal transduction pathways and protein degradation machinery inside the cell and sets stage for gaining further insights into regulation of protein degradation.

Two overrun phenomena and their effects on fusion yield in the Coulomb explosion of heteronuclear clusters

Two overrun effects in the Coulomb explosion dynamics of heteronuclear clusters have been investigated theoretically by the use of a simplified electrostatic model. When the charge-to-mass ratio of light ions is higher than that of heavy ions, the light ions can overtake the heavy ions inside the cluster and acquire a higher kinetic energy. Further, if the charge density of the heavy ions is twice as high as that of the light ions, i.e. a proposed competitive parameter xi = rho BqB/rho AqA > 2, the inner light ions can overtake those light ions on the surface of the cluster and form a shock shell during the explosion, which might drive the intracluster collision and fusion of the light ions. Different regimes of nuclear fusion are discussed and the corresponding neutron yields are estimated. Our analysis indicates that the probability of intracluster fusion is quite low even if deuterated heteronuclear clusters such as (DI)(n) with large size and high competitive parameter are employed. However, heteronuclear clusters are still a better candidate compared with homonuclear clusters for enhancing the total intercluster fusion yield because both a higher energy region and a higher proportion of deuterons distributing in the energy region can be created in the deuterated heteronuclear clusters.

Structural geology of the region between Pacoima and Little Tujunga Canyons, San Gabriel Mountains, California

An area of about 25 square miles in the western part of the San Gabriel Mountains was mapped on a scale of 1000 feet to the inch. Special attention was given to the structural geology, particularly the relations between the different systems of faults, of which the San Gabriel fault system and the Sierra Madre fault system are the most important ones. The present distribution and relations of the rocks suggests that the southern block has tilted northward against a more stable mass of old rocks which was raised up during a Pliocene or post-Pliocene orogeny. It is suggested that this northward tilting of the block resulted in the group of thrust faults which comprise the Sierra Madre fault system. It is show that this hypothesis fits the present distribution of the rocks and occupies a logical place in the geologic history of the region as well or better than any other hypothesis previously offered to explain the geology of the region.

Decarbonising urban transportation

18 p.

Plasma loop and strapping field dynamics: reproducing solar eruptions in the laboratory

Coronal mass ejections (CMEs) are dramatic eruptions of large, plasma structures from the Sun. These eruptions are important because they can harm astronauts, damage electrical infrastructure, and cause auroras. A mysterious feature of these eruptions is that plasma-filled solar flux tubes first evolve slowly, but then suddenly erupt. One model, torus instability, predicts an explosive-like transition from slow expansion to fast acceleration, if the spatial decay of the ambient magnetic field exceeds a threshold.

We create arched, plasma filled, magnetic flux ropes similar to CMEs. Small, independently-powered auxiliary coils placed inside the vacuum chamber produce magnetic fields above the decay threshold that are strong enough to act on the plasma. When the strapping field is not too strong and not too weak, expansion force build up while the flux rope is in the strapping field region. When the flux rope moves to a critical height, the plasma accelerates quickly, corresponding to the observed slow-rise to fast-acceleration of most solar eruptions. This behavior is in agreement with the predictions of torus instability.

Historically, eruptions have been separated into gradual CMEs and impulsive CMEs, depending on the acceleration profile. Recent numerical studies question this separation. One study varies the strapping field profile to produce gradual eruptions and impulsive eruptions, while another study varies the temporal profile of the voltage applied to the flux tube footpoints to produce the two eruption types. Our experiment reproduced these different eruptions by changing the strapping field magnitude, and the temporal profile of the current trace. This suggests that the same physics underlies both types of CME and that the separation between impulsive and gradual classes of eruption is artificial.

Study of the transmitter antenna gain for intersatellite laser communications

The gain of a transmitter for intersatellite communications is closely related to the performance of all the links. We calculate the transmitter telescope's gain with the help of the rigorous scalar diffraction theory and equivalent optical layout method. Furthermore, a comparison is performed with the conventional imaging method. The results show that the stop inside the telescope can affect the gain of the telescope. Finally, the gain is calculated under the condition of the aberrations. We find that different aberrations cause different effects. (C) 2006 Society of Photo-Optical Instrumentation Engineers.

Study of the transmitter antenna gain for intersatellite laser communications

The gain of a transmitter for intersatellite communications is closely related to the performance of all the links. We calculate the transmitter telescope's gain with the help of the rigorous scalar diffraction theory and equivalent optical layout method. Furthermore, a comparison is performed with the conventional imaging method. The results show that the stop inside the telescope can affect the gain of the telescope. Finally, the gain is calculated under the condition of the aberrations. We find that different aberrations cause different effects. (C) 2006 Society of Photo-Optical Instrumentation Engineers.

Improved uniformity of target illumination by combining a lens array and the technique of spectral dispersion

A scheme using a lens array and the technique of spectral dispersion is presented to improve target illumination uniformity in laser produced plasmas. Detailed two-dimensional simulation shows that a quasi-near-field target pattern, of steeper edges and without side lobes, is achieved with a lens array, while interference stripes inside the pattern are smoothed out by the use of the spectral dispersion technique. Moving the target slightly from the exact focal plane of the principal focusing lens can eliminate middle-scale-length intensity fluctuation further. Numerical results indicate that a well-irradiated laser spot with small nonuniformity and great energy efficiency can be obtained in this scheme. (c) 2007 American Institute of Physics.

Two-dimensional performance of uniform irradiation with the use of an edge-softened lens array and spectral dispersion

A lens array composed of edge-softened elements is used to improve on-target irradiation uniformity in the Shenguang II Laser Facility, with which a Fresnel pattern of suppressed diffraction peaks is obtained. Additional uniformity can be reached by reducing short-wavelength interference speckles inside the pattern when the technique of smoothing by spectral dispersion is also used. Two-dimensional performance of irradiation is simulated and the results indicate that a pattern of steeper edges and a flat top can be achieved with this joint technique. (c) 2007 Optical Society of America.

A bacia do Rio Imboassu, no município de São Gonçalo (RJ) como unidade viabilizadora da educação ambiental no contexto do planejamento e gestão ambiental

Os problemas ambientais são cada vez mais comuns e de magnitudes e escalas variadas, atingindo principalmente as áreas urbanas. Poluição atmosférica e dos corpos dágua, deslizamentos de encostas e enchentes são alguns dessas situações adversas. São Gonçalo não foge á regra. Localizada na Região Metropolitana do Rio de Janeiro, essa cidade sofre com as situações exemplificadas, principalmente em relação a degradação dos rios urbanos e as enchentes que acometem algumas áreas do município. Partindo-se da problemática exposta, a presente pesquisa tem como objetivo principal promover uma discussão teórica e reflexiva sobre a importância da efetivação da educação ambiental com foco na bacia hidrográfica urbana, tomando-se a bacia hidrográfica do rio Imboassú no município de São Gonçalo (RJ), como recorte. A fundamentação metodológica deste trabalho está pautada na análise ambiental que prioriza a participação pública e a educação ambiental no processo de planejamento e gestão, visando minimizar as diversas situações de desequilíbrio e degradação que acometem a bacia. A operacionalização deu-se do seguinte modo: análise inicial a partir da sistematização de estudos diagnósticos e de consultas de documentos, relatórios, dentre outros de órgãos governamentais; análise intermediária a partir da observação em campo do atual estado de degradação da bacia hidrográfica do rio Imboassú e da identificação do conjunto de políticas públicas vigentes; análise integrada análise dos estudos diagnósticos, análise da situação atual da bacia (in loco e documental), análise final com identificação de lacunas de gestão e propostas viáveis no âmbito da educação ambiental. Verificou-se que essa bacia está urbanizada e os rios que a compõem descaracterizados e poluídos, com histórico de enchentes. Falta de ordenamento urbano, degradação ambiental e descaso do poder público são alguns problemas enfrentados pelos moradores dessa bacia, resultando na falta de ordenamento territorial urbano e falta de qualidade de vida da população. Diante do exposto, fica claro a necessidade de se criar mecanismos que amenizem essa degradação e tragam equilíbrio ao funcionamento e dinâmica da bacia, influenciando positivamente o dia-a-dia da população residente em seu interior. Um eficiente caminho para promover essa mudança é a educação ambiental como ferramenta para a transformação da sociedade, conscientizando-a de seu papel participativo e modificando a maneira como esta se relaciona com o meio ambiente. Verificou-se que na prática, na bacia do Imboassú, o poder público não efetiva os pressupostos presentes, tanto na legislação ambiental, quanto no plano diretor municipal e alguns órgãos (CEDAE, INEA, SEMMA) não atuam de forma satisfatória. Logo, levantar a discussão da importância da Educação Ambiental como política pública a ser promovida pelos gestores municipais e apontar para a participação e atuação da sociedade de forma crítica na estruturação do espaço urbano é de extrema importância para promover melhoria na relação sociedade meio ambiente.

Métodos espectronodais para cálculos de transporte de partículas neutras com fonte fixa na formulação de ordenadas discretas e multigrupo de energia

Um método espectronodal é desenvolvido para problemas de transporte de partículas neutras de fonte fixa, multigrupo de energia em geometria cartesiana na formulação de ordenadas discretas (SN). Para geometria unidimensional o método espectronodal multigrupo denomina-se método spectral Greens function (SGF) com o esquema de inversão nodal (NBI) que converge solução numérica para problemas SN multigrupo em geometria unidimensional, que são completamente livre de erros de truncamento espacial para ordem L de anisotropia de espalhamento desde que L < N. Para geometria X; Y o método espectronodal multigrupo baseia-se em integrações transversais das equações SN no interior dos nodos de discretização espacial, separadamente nas direções coordenadas x e y. Já que os termos de fuga transversal são aproximados por constantes, o método nodal resultante denomina-se SGF-constant nodal (SGF-CN), que é aplicado a problemas SN multigrupo de fonte fixa em geometria X; Y com espalhamento isotrópico. Resultados numéricos são apresentados para ilustrar a eficiência dos códigos SGF e SGF-CN e a precisão das soluções numéricas convergidas em cálculos de malha grossa.

Formar almas, plasmar corações, dirigir vontades: o projeto educacional das Filhas da Caridade da Sociedade São Vicente de Paulo (1898-1905)

Foi analisado, nesta pesquisa, as condições de possibilidades para criação do Colégio Nossa Senhora das Dores na cidade de São João del-Rei, Minas Gerais. Fundado em1898 pelas Filhas da Caridade, iniciou suas atividades com o ensino voltado para um público exclusivamente feminino. O prédio foi construído para abrigar cem alunas internas e tantas outras externas para a formação primária, "ginasial", secundária e o Curso Normal, tendo funcionado em regime de internato, semi-internato e externato. Ao considerar os diferentes estratos de formação para os quais a escola se voltou, enfatizei o exame no nível da formação docente, buscando compreender as relações entre sociedade e educação escolar religiosa e suas articulações com os modelos educacionais difundidos à época. Para tanto, analisei os dispositivos disciplinares destinados a institucionalizar um programa, tendo como princípio a difusão da doutrina cristã associada à formação docente praticada no Colégio no início do seu funcionamento. No projeto de construção é possível observar a preocupação dos idealizadores em reafirmar um modelo escolar de formação do magistério em meio ao debate da qualificação docente em que diferentes modelos de formação estiveram em pauta. Em função do entrelaçamento dos ambientes urbanos e escolares produzidos no final do século XIX e início do XX, as remodelações de cidade fizeram emergir, em São João del-Rei, um edifício exemplar, representado na configuração do ecletismo mineiro. Nessa investigação, os saberes e tempos escolares se tornaram imprescindíveis para proceder à análise da criação e funcionamento da instituição examinada. Da mesma forma, o exame da instituição foi perseguido à medida que essas preocupações apareceram associadas, visto que os lugares e as práticas que os instituíram não podem ser deslocados do que se conseguiu produzir. O estudo enfocou o período entre a criação da instituição (1898) até 1905, ano em que o Curso Normal do CNSD foi equiparado aos cursos das escolas normais oficiais do Estado. Foi baseado na análise de leis, periódicos, relatórios de presidentes de província/estado, normas e estatutos da congregação e de uma vasta documentação localizada no CNSD. Ao entrecruzar essas fontes, a presente pesquisa buscou problematizar as dimensões de governo contidas nos planos da Igreja Católica, em termos gerais, e da Congregação da Missão de São Vicente de Paulo e das Filhas da Caridade, em específico. Pode-se dizer que o CNSD, por meio das Filhas da Caridade da Sociedade São Vicente de Paulo, deixou marcas em São João del-Rei como uma escola brasileira católica cristã, o que reforça a larga tradição de fazer da escola um instrumento de afirmação da fé. Para tal, muito contribuiu o novo estatuto jurídico adquirido em 1905, ampliando a legitimidade conquistada pelo CNSD, constituindo um sinal de forças das "irmãs" na cidade, assim como na eficiência de estratégias e de efetividade da ação educativa protagonizada pelas vicentinas em São João del-Rei. Nessa perspectiva, é possível afirmar que o projeto expansionista do catolicismo romanizado buscou alargar seu raio de ação em diversos locais, seja no interior de uma cidade mineira, seja no Brasil ou em outros países.