Energy-driven subunit rotation at the interface between subunit a and the c oligomer in the FO sector of Escherichia coli ATP synthase

Subunit rotation within the F1 catalytic sector of the ATP synthase has been well documented, identifying the synthase as the smallest known rotary motor. In the membrane-embedded FO sector, it is thought that proton transport occurs at a rotor/stator interface between the oligomeric ring of c subunits (rotor) and the single-copy a subunit (stator). Here we report evidence for an energy-dependent rotation at this interface. FOF1 was expressed with a pair of substituted cysteines positioned to allow an intersubunit disulfide crosslink between subunit a and a c subunit [aN214C/cM65C; Jiang, W. & Fillingame, R. H. (1998) Proc. Natl. Acad. Sci. USA 95, 6607–6612]. Membranes were treated with N,N′-dicyclohexyl-[14C]carbodiimide to radiolabel the D61 residue on less than 20% of the c subunits. After oxidation to form an a–c crosslink, the c subunit properly aligned to crosslink to subunit a was found to contain very little 14C label relative to other members of the c ring. However, exposure to MgATP before oxidation significantly increased the radiolabel in the a–c crosslink, indicating that a different c subunit was now aligned with subunit a. This increase was not induced by exposure to MgADP/Pi. Furthermore, preincubation with MgADP and azide to inhibit F1 or with high concentrations of N,N′-dicyclohexylcarbodiimide to label most c subunits prevented the ATP effect. These results provide evidence for an energy-dependent rotation of the c ring relative to subunit a.

Rotation of subunits during catalysis by Escherichia coli F1-ATPase.

During oxidative and photo-phosphorylation, F0F1-ATP synthases couple the movement of protons down an electrochemical gradient to the synthesis of ATP. One proposed mechanistic feature that has remained speculative is that this coupling process requires the rotation of subunits within F0F1. Guided by a recent, high-resolution structure for bovine F1 [Abrahams, J. P., Leslie, A. G., Lutter, R. & Walker, J. E. (1994) Nature (London) 370, 621-628], we have developed a critical test for rotation of the central gamma subunit relative to the three catalytic beta subunits in soluble F1 from Escherichia coli. In the bovine F1 structure, a specific point of contact between the gamma subunit and one of the three catalytic beta subunits includes positioning of the homolog of E. coli gamma-subunit C87 (gamma C87) close to the beta-subunit 380DELSEED386 sequence. A beta D380C mutation allowed us to induce formation of a specific disulfide bond between beta and gamma C87 in soluble E. coli F1. Formation of the crosslink inactivated beta D380C-F1, and reduction restored full activity. Using a dissociation/reassembly approach with crosslinked beta D380C-F1, we incorporated radiolabeled beta subunits into the two noncrosslinked beta-subunit positions of F1. After reduction of the initial nonradioactive beta-gamma crosslink, only exposure to conditions for catalytic turnover results in similar reactivities of unlabeled and radiolabeled beta subunits with gamma C87 upon reoxidation. The results demonstrate that gamma subunit rotates relative to the beta subunits during catalysis.

Magnetic activity and differential rotation in the young Sun-like stars KIC 7985370 and KIC 7765135

Aims. We present a detailed study of the two Sun-like stars KIC 7985370 and KIC 7765135, to determine their activity level, spot distribution, and differential rotation. Both stars were previously discovered by us to be young stars and were observed by the NASA Kepler mission. Methods. The fundamental stellar parameters (vsini, spectral type, T_eff, log g, and [Fe/H]) were derived from optical spectroscopy by comparison with both standard-star and synthetic spectra. The spectra of the targets allowed us to study the chromospheric activity based on the emission in the core of hydrogen Hα and Ca ii infrared triplet (IRT) lines, which was revealed by the subtraction of inactive templates. The high-precision Kepler photometric data spanning over 229 days were then fitted with a robust spot model. Model selection and parameter estimation were performed in a Bayesian manner, using a Markov chain Monte Carlo method. Results. We find that both stars are Sun-like (of G1.5 V spectral type) and have an age of about 100–200 Myr, based on their lithium content and kinematics. Their youth is confirmed by their high level of chromospheric activity, which is comparable to that displayed by the early G-type stars in the Pleiades cluster. The Balmer decrement and flux ratio of their Ca ii-IRT lines suggest that the formation of the core of these lines occurs mainly in optically thick regions that are analogous to solar plages. The spot model applied to the Kepler photometry requires at least seven persistent spots in the case of KIC 7985370 and nine spots in the case of KIC 7765135 to provide a satisfactory fit to the data. The assumption of the longevity of the star spots, whose area is allowed to evolve with time, is at the heart of our spot-modelling approach. On both stars, the surface differential rotation is Sun-like, with the high-latitude spots rotating slower than the low-latitude ones. We found, for both stars, a rather high value of the equator-to-pole differential rotation (dΩ ≈ 0.18 rad d^-1), which disagrees with the predictions of some mean-field models of differential rotation for rapidly rotating stars. Our results agree instead with previous works on solar-type stars and other models that predict a higher latitudinal shear, increasing with equatorial angular velocity, that can vary during the magnetic cycle.

Chromospheric activity and rotation of FGK stars in the solar vicinity. An estimation of the radial velocity jitter

Context. Chromospheric activity produces both photometric and spectroscopic variations that can be mistaken as planets. Large spots crossing the stellar disc can produce planet-like periodic variations in the light curve of a star. These spots clearly affect the spectral line profiles, and their perturbations alter the line centroids creating a radial velocity jitter that might “contaminate” the variations induced by a planet. Precise chromospheric activity measurements are needed to estimate the activity-induced noise that should be expected for a given star. Aims. We obtain precise chromospheric activity measurements and projected rotational velocities for nearby (d ≤ 25 pc) cool (spectral types F to K) stars, to estimate their expected activity-related jitter. As a complementary objective, we attempt to obtain relationships between fluxes in different activity indicator lines, that permit a transformation of traditional activity indicators, i.e., Ca II H & K lines, to others that hold noteworthy advantages. Methods. We used high resolution (~50 000) echelle optical spectra. Standard data reduction was performed using the IRAF ECHELLE package. To determine the chromospheric emission of the stars in the sample, we used the spectral subtraction technique. We measured the equivalent widths of the chromospheric emission lines in the subtracted spectrum and transformed them into fluxes by applying empirical equivalent width and flux relationships. Rotational velocities were determined using the cross-correlation technique. To infer activity-related radial velocity (RV) jitter, we used empirical relationships between this jitter and the R’_HK index. Results. We measured chromospheric activity, as given by different indicators throughout the optical spectra, and projected rotational velocities for 371 nearby cool stars. We have built empirical relationships among the most important chromospheric emission lines. Finally, we used the measured chromospheric activity to estimate the expected RV jitter for the active stars in the sample.

Estudo da topologia de motores de fluxo axial de excitação híbrida com foco na utilização em tração elétrica.

Os motores de corrente contínua convencionais são muito bem conhecidos pela sua robustez e pelo seu alto nível de controlabilidade, alem do fato de possibilitarem a operação na região de enfraquecimento de campo (modo motor), quando esta situação se fizer necessária. Por estas características, as máquinas de corrente contínua ainda são empregadas nos dias atuais em nichos específicos de utilização. Não obstante, a máquina c.c. apresenta algumas desvantagens, principalmente a intensiva e dispendiosa manutenção eletromecânica necessária para sua operação. Como opção de sanar este problema, surgiram na década de 60, as máquinas elétricas de corrente contínua sem escovas (brushless) com excitação por ímãs permanentes de fluxo trapezoidal. O problema destas máquinas se deve justamente a impossibilidade da variação de fluxo de excitação uma vez que são produzidos puramente pelos ímãs. Sendo assim, este trabalho tem como propósito, o estudo de topologias diferenciadas da máquina elétrica, através de um circuito magnético não convencional para aplicação e utilização em sistemas de tração elétrica para operação na região de enfraquecimento de campo através da variação do fluxo resultante no entreferro. Como objeto de estudo, foi focada a topologia de fluxo axial com excitação híbrida, ou seja, dupla excitação (excitação a ímãs permanentes e excitação elétrica). Para o projeto da topologia proposta, nesta tese, adicionalmente ao método analítico, foram realizadas simulações computacionais para a comparação e refinamento dos resultados das grandezas eletromagnéticas da máquina.

Modelagem estocástica da dispersão axial: aplicação em um reator tubular de polimerização.

Reatores tubulares de polimerização podem apresentar um perfil de velocidade bastante distorcido. Partindo desta observação, um modelo estocástico baseado no modelo de dispersão axial foi proposto para a representação matemática da fluidodinâmica de um reator tubular para produção de poliestireno. A equação diferencial foi obtida inserindo a aleatoriedade no parâmetro de dispersão, resultando na adição de um termo estocástico ao modelo capaz de simular as oscilações observadas experimentalmente. A equação diferencial estocástica foi discretizada e resolvida pelo método Euler-Maruyama de forma satisfatória. Uma função estimadora foi desenvolvida para a obtenção do parâmetro do termo estocástico e o parâmetro do termo determinístico foi calculado pelo método dos mínimos quadrados. Uma análise de convergência foi conduzida para determinar o número de elementos da discretização e o modelo foi validado através da comparação de trajetórias e de intervalos de confiança computacionais com dados experimentais. O resultado obtido foi satisfatório, o que auxilia na compreensão do comportamento fluidodinâmico complexo do reator estudado.

Choroidal Thickness And Volume In Healthy Young Whites And Their Relationship With Axial Length, Ammetropy And Sex

Purpose: To evaluate choroidal thickness in young subjects using Enhanced Depth Imaging Spectral Domain Optical Coherence Tomography (EDI SD-OCT) describing volume differences between all the defined areas of the Early Treatment Diabetic Retinopathy Study (ETDRS). Design: Prospective, clinical study. Methods: Seventy-nine eyes of 95 healthy, young (23.8±3.2years), adult volunteers were prospectively enrolled. Manual choroidal segmentation on a 25-raster horizontal scan protocol was performed. The measurements of the nine subfields defined by the ETDRS were evaluated. Results: Mean subfoveal choroidal thickness was 345.67±81.80μm and mean total choroidal volume was 8.99±1.88mm3. Choroidal thickness and volume were higher at the superior and temporal areas compared to inferior and nasal sectors of the same diameter respectively. Strong correlations between subfoveal choroidal thickness and axial length (AL) and myopic refractive error were obtained, r = -0.649, p<0.001 and r = 0.473, p<0.001 respectively. Emmetropic eyes tended to have thicker subfoveal choroidal thickness (381.94±79.88μm versus 307.04±64.91μm) and higher total choroidal volume than myopic eyes (9.80± 1.87mm3 versus 8.14±1.48mm3). The estimation of the variation of the subfoveal choroidal thickness with the AL was-43.84μm/mm. In the myopic group, the variation of the subfoveal choroidal thickness with the myopic refractive error was -10.45μm/D. Conclusions: This study establishes for the first time a normal database for choroidal thickness and volume in young adults. Axial length, and myopic ammetropy are highly associated with choroidal parameters in healthy subjects. EDI SD-OCT exhibited a high degree of intraobserver and interobserver repeatability.

Cyclobutadiene automerization and rotation of ethylene: Energetics of the barriers by using Spin-polarized wave functions

Spin-projected spin polarized Møller–Plesset and spin polarized coupled clusters calculations have been made to estimate the cyclobutadiene automerization, the ethylene torsion barriers in their ground state, and the gap between the singlet and triplet states of ethylene. The results have been obtained optimizing the geometries at MP4 and/or CCSD levels, by an extensive Gaussian basis set. A comparative analysis with more complex calculations, up to MP5 and CCSDTQP, together with others from the literature, have also been made, showing the efficacy of using spin-polarized wave functions as a reference wave function for Møller–Plesset and coupled clusters calculations, in such problems.

Evidence of Intracrystalline Mesostructured Porosity in Zeolites by Advanced Gas Sorption, Electron Tomography and Rotation Electron Diffraction

The small size of micropores (typically <1 nm) in zeolites causes slow diffusion of reactant and product molecules in and out of the pores and negatively impacts the product selectivity of zeolite based catalysts, for example, fluid catalytic cracking (FCC) catalysts. Size-tailored mesoporosity was introduced into commercial zeolite Y crystals by a simple surfactant-templating post-synthetic mesostructuring process. The resulting mesoporous zeolite Y showed significantly improved product selectivity in both laboratory testing and refinery trials. Advanced characterization techniques such as electron tomography, three-dimensional rotation electron diffraction, and high resolution gas adsorption coupled with hysteresis scanning and density functional theory, unambiguously revealed the intracystalline nature and connectivity of the introduced mesopores. They can be considered as molecular highways that help reactant and product molecules diffuse quickly to and away from the catalytically active sites within the zeolite crystals and, thus, shift the selectivity to favor the production of more of the valuable liquid fuels at reduced yields of coke and unconverted feed.

Relationship among Corneal Biomechanics, Refractive Error, and Axial Length

Purpose: To evaluate the relationship between different ocular and corneal biomechanical parameters in emmetropic and ametropic healthy white children. Methods: This study included 293 eyes of 293 healthy Spanish children (135 boys and 158 girls), ranging in age from 6 to 17 years. Subjects were divided according to the refractive error: control (emmetropia, 99 children), myopia (100 children), and hyperopia (94 children) groups. In all cases, corneal hysteresis (CH) and corneal resistance factor (CRF) were evaluated with the Ocular Response Analyzer system. Axial length (AL) and mean corneal power were also measured by partial coherence interferometry (IOLMaster), and central corneal thickness (CCT) and anterior chamber depth were measured by anterior segment optical coherence tomography (Visante). Results: Mean (±SD) CH and CRF were 12.12 (±1.71) and 12.30 (±1.89) mm Hg, respectively. Mean (±SD) CCT was 542.68 (±37.20) μm and mean (±SD) spherical equivalent was +0.14 (±3.41) diopters. A positive correlation was found between CH and CRF (p < 0.001), and both correlated as well with CCT (p < 0.0001). Corneal resistance factor was found to decrease with increasing age (p = 0.01). Lower levels of CH were associated with longer AL and more myopia (p < 0.001 and p = 0.001, respectively). Higher values of CH were associated with increasing hyperopia. Significant differences in CH were found between emmetropic and myopic groups (p < 0.001) and between myopic and hyperopic groups (p = 0.011). There were also significant differences in CRF between emmetropic and myopic groups (p = 0.02). Multiple linear regression analysis showed that lower CH and CRF significantly associated with thinner CCT, longer AL, and flatter corneal curvature. Conclusions: The Ocular Response Analyzer corneal biomechanical properties seem to be compromised in myopia from an early age, especially in high myopia.

The Gaia-ESO Survey: Empirical determination of the precision of stellar radial velocities and projected rotation velocities

Context. The Gaia-ESO Survey (GES) is a large public spectroscopic survey at the European Southern Observatory Very Large Telescope. Aims. A key aim is to provide precise radial velocities (RVs) and projected equatorial velocities (vsini) for representative samples of Galactic stars, which will complement information obtained by the Gaia astrometry satellite. Methods. We present an analysis to empirically quantify the size and distribution of uncertainties in RV and vsini using spectra from repeated exposures of the same stars. Results. We show that the uncertainties vary as simple scaling functions of signal-to-noise ratio (S/N) and vsini, that the uncertainties become larger with increasing photospheric temperature, but that the dependence on stellar gravity, metallicity and age is weak. The underlying uncertainty distributions have extended tails that are better represented by Student’s t-distributions than by normal distributions. Conclusions. Parametrised results are provided, which enable estimates of the RV precision for almost all GES measurements, and estimates of the vsini precision for stars in young clusters, as a function of S/N, vsini and stellar temperature. The precision of individual high S/N GES RV measurements is 0.22–0.26 km s-1, dependent on instrumental configuration.

Earth’s Rotation: A Challenging Problem in Mathematics and Physics

A suitable knowledge of the orientation and motion of the Earth in space is a common need in various fields. That knowledge has been ever necessary to carry out astronomical observations, but with the advent of the space age, it became essential for making observations of satellites and predicting and determining their orbits, and for observing the Earth from space as well. Given the relevant role it plays in Space Geodesy, Earth rotation is considered as one of the three pillars of Geodesy, the other two being geometry and gravity. Besides, research on Earth rotation has fostered advances in many fields, such as Mathematics, Astronomy and Geophysics, for centuries. One remarkable feature of the problem is in the extreme requirements of accuracy that must be fulfilled in the near future, about a millimetre on the tangent plane to the planet surface, roughly speaking. That challenges all of the theories that have been devised and used to-date; the paper makes a short review of some of the most relevant methods, which can be envisaged as milestones in Earth rotation research, emphasizing the Hamiltonian approach developed by the authors. Some contemporary problems are presented, as well as the main lines of future research prospected by the International Astronomical Union/International Association of Geodesy Joint Working Group on Theory of Earth Rotation, created in 2013.

Avaliação da composição corporal e do estado nutricional em doentes com neoplasias gastrointestinais : bioimpedância elétrica e tomografia axial computorizada, que correlação?

Tese de mestrado, Nutrição Clínica, Faculdade de Medicina, Universidade de Lisboa, 2015