Pivoting between Calmodulin Lobes Triggered by Calcium in the Kv7.2/Calmodulin Complex

Kv7.2 (KCNQ2) is the principal molecular component of the slow voltage gated M-channel, which strongly influences neuronal excitability. Calmodulin (CaM) binds to two intracellular C-terminal segments of Kv7.2 channels, helices A and B, and it is required for exit from the endoplasmic reticulum. However, the molecular mechanisms by which CaM controls channel trafficking are currently unknown. Here we used two complementary approaches to explore the molecular events underlying the association between CaM and Kv7.2 and their regulation by Ca2+. First, we performed a fluorometric assay using dansylated calmodulin (D-CaM) to characterize the interaction of its individual lobes to the Kv7.2 CaM binding site (Q2AB). Second, we explored the association of Q2AB with CaM by NMR spectroscopy, using N-15-labeled CaM as a reporter. The combined data highlight the interdependency of the N- and C-lobes of CaM in the interaction with Q2AB, suggesting that when CaM binds Ca2+ the binding interface pivots between the N-lobe whose interactions are dominated by helix B and the C-lobe where the predominant interaction is with helix A. In addition, Ca2+ makes CaM binding to Q2AB more difficult and, reciprocally, the channel weakens the association of CaM with Ca2+.

Reflex impairment as a measure of vitality and survival potential of Atlantic cod (Gadus morhua)

A ssur ing the v itality and survival potential of live-caught Atlantic cod (Gadus morhua) is important for improving the sorting of fish before net penning operations designed to hold fish for growth and later market. When Atlantic cod are captured by Danish seine, the most commonly used fishing gear for live-caught fish, they undergo stressors such as forced swimming, net abrasion, and air exposure. Laboratory experiments (at an air temperature of 9°C and water temperature of 8°C) were conducted with the aim of constructing a RAMP (reflex action mortality predictor) curve for prediction of vitality and survival potential in Atlantic cod captured in Danish seines, by varying the levels of these stressors. Atlantic cod exposed to increased duration in air (5–20 min) showed increased reflex impairment and mortality, with 75% mortality at 10 minutes of air exposure. Forced swimming in combination with net abrasion and air exposure did not increase reflex impairment or mortality above that associated with air exposure alone. The Atlantic cod RAMP curves indicated that fish with reflex impairment less than 50% would not show mortality and would likely recover from capture stress.

Identificação da formação de titanato de bário a partir de mistura reacional calcinada em diferentes temperaturas via difração de raios X, espectroscopia fotoacústica e análise térmica

Os materiais ferroelétricos têm sido utilizados em muitas áreas da tecnologia e da ciência, pois possuem um grande número de aplicações, como: sensores; transdutores; capacitores; dispositivos ópticos; dentre outras. A busca por novos materiais cerâmicos ferroelétricos tem sido grande. Um dos materiais cerâmicos ferroelétricos mais estudados é o titanato de bário (BT). São vários os métodos de produção e caracterização do titanato de bário. Neste trabalho, pós cerâmicos de titanato de bário foram obtidos por reação do estado sólido a partir de misturas reacionais calcinadas em diferentes temperaturas entre 400C e 900C. Foram três as misturas reacionais: não dopadas; dopadas com 1%; e dopadas com 5% de dióxido de cério (CeO2). A identificação da formação do BT, nos pós cerâmicos produzidos, foi feita a partir de três técnicas de caracterização: difração de raios X (DRX); espectroscopia fotoacústica (PAS); e técnicas de análise térmica. Com a técnica DRX, difratogramas mostraram que a plena formação do titanato de bário ocorreu a partir da temperatura de calcinação de 700C. Para a amostra não dopada com cério e calcinada a 800C, houve deslocamento de todos os picos de difração. Nas amostras dopadas com dióxido de cério houve deslocamento de todos os picos de difração, em relação as amostras não dopadas. Observou-se também que nas amostras dopadas com 5% de CeO2, e calcinadas a 700C e 800C, resíduos de dióxido de cério foram observados nos difratogramas. Com a técnica PAS, espectros de absorção foram obtidos. Foi possível observar uma grande diferença de absorção da amostra calcinada a 600 e 630C, indicando a formação do titanato de bário a partir da temperatura de 630C, nas amostras sem a dopagem dióxido de cério. Houve um alargamento nas bandas de absorção a partir da temperatura de 600C, quando o dióxido de cério entrou na matriz. Foi também possível determinar as energias de band-gap das amostras utilizando o método de Tauc. Com as técnicas de análise térmica, em especial através da técnica termogravimétrica (TG/DTG), foi comprovado que até 400C não havia formação de titanato de bário. Visto que nesta temperatura de calcinação houve a maior perda de massa durante a rampa de aquecimento. O início da formação do titanato de bário foi observado a partir da temperatura de calcinação de 500C, assim como nas técnicas DRX e PAS. Portanto, com os resultados apresentados, foi demonstrada a identificação da formação do titanato de bário nas misturas reacionais calcinadas, com auxílio das potencialidades das três técnicas utilizadas.

Efeito da estimulação transcraniana por corrente contínua sobre o desempenho físico: análise das respostas cerebrais, metabólicas e perceptivas durante e após exercícios aeróbio e de força

Estimulação transcraniana por corrente contínua (ETCC) sobre áreas corticais pré-selecionadas, tem aumentado o desempenho físico de diferentes populações. Porém, lacunas persistem no tocante aos mecanismos subjacentes à estes efeitos. Assim, a presente tese objetivou: a) investigar os efeitos da ETCC anódica (aETCC) e placebo (Sham) no córtex motor (CM) de indivíduos saudáveis sobre o desempenho de força máxima; b) comparar os efeitos da ETCC sobre a produção de força máxima e estabilidadade da força durante exercícios máximo e submáximo em sujeitos hemiparéticos e saudáveis; c) investigar o efeito da ETCC sobre a conectividade funcional inter-hemisférica (coerência eletroencefalográfica cEEG) do córtex pré-frontal (CPF), desempenho aeróbio e dispêndio energético (EE) durante e após exercício máximo e submáximo. No 1 estudo, 14 adultos saudáveis executaram 2 sessões de exercício máximo de força (EMF) dos músculos flexores e extensores do joelho dominante (3 séries de 10 rep máximas), precedidos por aETCC ou Sham (2mA; 20 mim). aETCC não foi capaz de aumentar o trabalho total e pico de torque (PT), resistência à fadiga ou atividade eletromiográfica durante o EMF. No 2 estudo, 10 hemiparéticos e 9 sujeitos saudáveis receberam aETCC e Sham no CM. O PT e a estabilidade da força (coeficiente de variação - CV) foram avaliados durante protocolo máximo e submáximo de extensão e flexão unilateral do joelho (1 série de 3 reps a 100% do PT e 2 séries de 10 reps a 50% do PT). Nenhuma diferença no PT foi observada nos dois grupos. Diminuições no CV foram obervadas durante a extensão (~25-35%, P<0.001) e flexão de joelho (~22-33%, P<0.001) após a aETCC comparada com Sham nos hemiparéticos, entretanto, somente o CV na extensão de joelhos diminuiu (~13-27%, P<0.001) nos saudáveis, o que sugere que aETCC pode melhorar o CV, mas não o PT em sujeitos hemiparéticos. No 3 estudo, 9 adultos saudáveis realizaram 2 testes incrementais máximos precedidos por aETCC ou Sham sobre o CPF com as respostas cardiorrespiratórias, percepção de esforço (PSE) e cEEG do CPF sendo monitoradas. O VO2 de pico (42.64.2 vs. 38.23.3 mL.kg.min-1; P=0,02), potência total (252.776.5 vs. 23773.3 W; P=0,05) e tempo de exaustão (531.1140 vs. 486.7115.3 seg; P=0,04) foram maiores após aETCC do que a Sham. Nenhuma diferença foi encontrada para FC e PSE em função da carga de trabalho (P>0,05). A cEEG do CPF aumentou após aETCC vs. repouso (0.700.40 vs. 0.380.05; P=0,001), mas não após Sham vs. repouso (0.360.49 vs. 0.330.50; P=0,06), sugerindo que a aETCC pode retardar a fadiga aumentando a conectividade funcional entre os hemisférios do CPF e desempenho aeróbio durante exercício exaustivo. No 4 estudo, o VO2 e EE foram avaliados em 11 adultos saudáveis antes, durante a aETCC ou Sham no CPF e 30 min após exercício aeróbio submáximo isocalórico (~200kcal). Diferenças não foram observadas no VO2 vs. repouso durante aETCC e Sham (P=0.95 e P=0.85). Porém, a associação entre exercício e aETCC aumentou em ~19% o EE após ao menos, 30 min de recuperação após exercício quando comparada a Sham (P<0,05).

Impacto do programa bolsa família na luta fundiária da comunidade quilombola da caiana dos crioulos: um estudo sobre cidadania

A presente tese analisa como o Programa Bolsa Família vem impactando a luta fundiária da Comunidade Quilombola de Caiana dos Crioulos, observando como isto tem afetado a construção e desenvolvimento da cidadania desta comunidade. Para isto, fazemos um estudo sobre o programa bolsa família, que foi criado com o fulcro de diminuir a fome, a pobreza e a desigualdade social, alcançando, paulatinamente, milhões de brasileiros, inclusive grande parcela dos quilombolas. Nesta seara percebemos que a vulnerabilidade social é uma marca presente nas comunidades quilombolas, principalmente devido à dificuldade de acesso a estas comunidades, além de uma prestação de serviços públicos não focados para sua identidade cultural. Atualmente, grande parcela dos quilombolas é beneficiária do Programa Bolsa Família e não estão conseguindo a autonomização deste programa por não haver uma política pública específica que possa estimular o desenvolvimento dos quilombolas, respeitando a cultura dos mesmos. Destacamos também que a concretização do pleito principal dos quilombolas, que é a titulação das suas terras, conforme previsto no artigo 68 do Ato de Disposições Constitucionais Transitórios da Constituição Federal de 1988, não vem sendo efetivada devido a grande burocracia para esse procedimento de titulação junto com interesses de grupos hegemônicos ligados a bancada ruralista. Para o desenvolvimento da pesquisa fizemos uso da abordagem qualitativa, concomitantemente foi feita uma pesquisa de campo, permitindo a observação direta dos fenômenos, preservando a singularidade do objeto social. Além disso, fizemos consultas a dados primários e secundários de órgãos públicos. Utilizamos como instrumentos de coleta de dados entrevistas semiestruturadas, optando pela utilização da análise de conteúdo, buscando-se analisar a realidade social num grau de profundidade que ultrapasse o senso comum. Seguimos uma das técnicas específicas da análise de conteúdo, que é a análise temática. Concluiu-se que a regularização fundiária definitiva junto com a concretização de políticas públicas específicas são o caminho para construção da cidadania quilombola, pois permitirá que haja uma segurança jurídica para os quilombolas. E apenas o programa bolsa família não pode ser um propiciador da quebra do ciclo intergeracional da pobreza que muito marca a história dos povos quilombolas, precisando ser pensado de forma interdisciplinar as portas de saída da pobreza, daí a necessidade de capabilities para que o quilombola possa usufruir de uma cidadania plena.

Impact of high-k dielectrics on breakdown performances of SiC and diamond Schottky diodes

This paper presents a comparison between SiC and diamond Schottky barrier diodes using the oxide ramp termination. The influences of the dielectric thickness and relative permittivity on the diode's electrical performance are investigated. Typical commercial drift layer parameters are used for this study. The extension of the space charge area throughout the drift region and the current distribution at breakdown are shown. The efficiency of the termination is also evaluated for both SiC and diamond diodes. © (2009) Trans Tech Publications, Switzerland.

Physiological ecology of juvenile chinook salmon (Oncorhynchus tshawytscha) at the southern end of their distribution, the San Francisco Estuary and Gulf of the Farallones, California

Juvenile chinook salmon, Oncorhynchus tshawytscha, from natal streams in California’s Central Valley demonstrated little estuarine dependency but grew rapidly once in coastal waters. We collected juvenile chinook salmon at locations spanning the San Francisco Estuary from the western side of the freshwater delta—at the confluence of the Sacramento and San Joaquin Rivers—to the estuary exit at the Golden Gate and in the coastal waters of the Gulf of the Farallones. Juveniles spent about 40 d migrating through the estuary at an estimated rate of 1.6 km/d or faster during their migration season (May and June 1997) toward the ocean. Mean growth in length (0.18 mm/d) and weight (0.02 g/d) was insignificant in young chinook salmon while in the estuary, but estimated daily growth of 0.6 mm/d and 0.5 g/d in the ocean was rapid (P≤0.001). Condition (K factor) declined in the estuary, but improved markedly in ocean fish. Total body protein, total lipid, triacylglycerols (TAG), polar lipids, cholesterol, and nonesterified fatty acids concentrations did not change in juveniles in the estuary, but total lipid and TAG were depleted in ocean juveniles. As young chinook migrated from freshwater to the ocean, their prey changed progressively in importance from invertebrates to fish larvae. Once in coastal waters, juvenile salmon appear to employ a strategy of rapid growth at the expense of energy reserves to increase survival potential. In 1997, environmental conditions did not impede development: freshwater discharge was above average and water temperatures were only slightly elevated, within the species’ tolerance. Data suggest that chinook salmon from California’s Central Valley have evolved a strong ecological propensity for a ocean-type life history. But unlike populations in the Pacific Northwest, they show little estuarine dependency and proceed to the ocean to benefit from the upwelling-driven, biologically productive coastal waters.

On-line topographic analysis in the SEM

A novel method for on-line topographic analysis of rough surfaces in the SEM has been investigated. It utilises a digital minicomputer configured to act as a programmable scan generator and automatic focusing unit. The computer is coupled to the microscope through digital-to-analogue converters which enable it to generate ramp waveforms allowing the beam to be scanned over a small sub-region of the field under program control. A further digital-to-analogue converter regulates the current supply to the objective lens of the microscope. The video signal is sampled by means of an analogue-to-digital converter and the resultant binary code stored in the computer's memory as an array of numbers describing relative image intensity. Computations based on the intensity gradient of the image allow the objective lens current to be found for the in-focus condition, which may be related to the working distance through a previous calibration experiment. The sensitivity of the method for detecting small height changes is theoretically of the order of 1 μm. In practice the operator specifies features of interest by means of a mobile spot cursor injected into the SEM display screen, or he may scan the specimen at sub-regions corresponding to pre-determined points on a regular grid defined by him. The operation then proceeds under program control. | A novel method for on-line topographic analysis of rough surfaces in the SEM has been investigated. It utilizes a digital minicomputer configured to act as a programmable scan generator and automatic focusing unit. A further digital-to-analog converter regulates the current supply to the objective lens of the microscope. The video signal is sampled by means of an analog-to-digital converter and the resultant binary code stored in the computer's memory as an array of numbers describing relative image intensity. The sensitivity of the method for detecting small height changes is theroretically of the order of 1 mu m.

Topographical analysis in the SEM using an automatic focusing technique

A novel technique for automated topographical analysis in the SEM has been investigated. It utilizes a 16-bit minicomputer arranged to act as an automatic focusing unit. The computer is coupled to the objective lens of the microscope, by means of a digital to analogue converter, and may regulate the excitation of the lens under program control. Further digital-to-analogue converters allow the computer to act as a programmable scan generator by applying ramp waveforms to the scan amplifiers, permitting the beam to be swept over a small sub-region of the field of interest. The video signal is sampled and applied to an analogue-to-digital converter; the resultant binary numbers are stored in computer memory as an array of values representing relative image intensities within a subregion. A differencing algorithm applied to the collected data allows the level of objective lens excitation to be found at which the sharpness of the image is optimized, and the excitation may be related to the working distance for that subregion through a previous calibration experiment. The sensitivity of the method for detecting small height changes is theoretically of the order of 1 μm.

Effect of blade tip geometry on the tip leakage flow in axial turbine cascades

The phenomenon of tip leakage has been studied in two linear cascades of turbine blades. The investigation includes an examination of the performance of the cascades with a variety of tip geometries. The effects of using plain tips, suction side squealers, and pressure side squealers are reported. Traverses of the exit flow field were made in order to determine the overall performance. A method of calculating the tip discharge coefficients for squealer geometries is put forward. In linking the tip discharge coefficient and cascade losses, a procedure for predicting the relative performance of tip geometries is developed. The model is used to examine the results obtained using the different tip treatments and to highlight the important aspects of the loss generation process.

Three-dimensional interactions in the rotor of an axial turbine

This article presents a study of the development of the three-dimensional flowfield within the rotor blades of a low-speed, large-scale axial flow turbine. Measurements have been performed in the rotating and stationary frames of reference. Time-mean data have been obtained using miniature five-hole pneumatic probes, whereas the unsteady development of the flow has been determined using three-axis subminiature hot-wire anemometers. Additional information is provided by the results of blade-surface flow-visualization experiments and surface-mounted hot-film anemometers. The development of the stator exit flow, as it passes through the rotor blades, is described. Unsteady data suggest that the presence of the rotor secondary and tip leakage flows restricts the region of unsteady interaction to near midspan when the stator wakes and secondary flows are adjacent to the suction surface. Surface-mounted hot-film data show that this affects the suction-side laminar-turbulent transition process.

Interpretation of measured profile losses in unsteady wake-turbine blade interaction studies

The interaction of wakes shed by a moving bladerow with a downstream bladerow causes unsteady flow. The meaning of the freestream stagnation pressure and stagnation enthalpy in these circumstances has been examined using simple analyses, measurements and CFD. The unsteady flow in question arises from the behaviour of the wakes as so-called negative-jets. The interactions of the negative-jets with the downstream blades lead to fluctuations in static pressure which in turn generate fluctuations in the stagnation pressure and stagnation enthalpy. It is shown that the fluctuations of the stagnation quantities created by unsteady effects within the bladerow are far greater than those within the incoming wake. The time-mean exit profiles of the stagnation pressure and stagnation enthalpy are affected by these large fluctuations. This phenomenon of energy separation is much more significant than the distortion of the time-mean exit profiles that is caused directly by the cross-passage transport associated with the negative-jet, as described by Kerrebrock and Mikolajczak. Finally, it is shown that if only time-averaged values of loss are required across a bladerow, it is nevertheless sufficient to determine the time-mean exit stagnation pressure.

An experimental and computational study of the formation of a streamwise shed vortex in a turbine stage

Shear layers shed by aircraft wings roll up into vortices. A similar, though far less common, phenomenon can occur in the wake of a turbomachine blade. This paper presents experimental data from a new single stage turbine that has been commissioned at the Whittle Laboratory. Two low aspect ratio stators have been tested with the same rotor row. Surface flow visualisation illustrates the extremely strong secondary flows present in both NGV designs. These secondary flows lead to conventional passage vortices but also to an intense vortex sheet which is shed from the trailing edge of the blades. Pneumatic probe traverse show how this sheet rolls up into a concentrated vortex in the second stator design, but not in the first. A simple numerical experiment is used to model the shear layer instability and the effects of trailing edge shape and exit yaw angle distribution are investigated. It is found that the latter has a strong influence on shear layer rollup: inhibiting the formation of a vortex downstream of NGV 1 but encouraging it behind NGV 2.

Wake, shock and potential field interactions in a 1.5 stage turbine: Part I: Vane-rotor and rotor-vane interaction

The composition of the time-resolved surface pressure field around a high-pressure rotor blade caused by the presence of neighboring blade rows was studied, with the individual effects of wake, shock and potential field interaction being determined. Two test geometries were considered: first, a high-pressure turbine stage coupled with a swan-necked diffuser exit duct; secondly, the same high-pressure stage but with a vane located in the downstream duct. Both tests were carried out at engine-representative Mach and Reynolds numbers. By comparing the results to time-resolved computational predictions of the flowfield, the accuracy with which the computation predicts blade interaction was determined. It was found that in addition to upstream vane-rotor and rotor-downstream vane interactions, a new interaction mechanism was found resulting from the interaction between the downstream vane's potential field and the upstream vane's trailing edge potential field and shock.

Wake, shock and potential field interactions in a 1.5 stage turbine: Part II: Vane-vane interaction and discussion of results

The composition of the time-resolved surface pressure field around a high-pressure rotor blade caused by the presence of neighboring blade rows was studied, with the individual effects of wake, shock and potential field interaction being determined. Two test geometries were considered: first, a high-pressure turbine stage coupled with a swan-necked diffuser exit duct; secondly, the same high-pressure stage but with a vane located in the downstream duct. Both tests were carried out at engine-representative Mach and Reynolds numbers. By comparing the results to time-resolved computational predictions of the flowfield, the accuracy with which the computation predicts blade interaction was determined. Evidence was obtained that for a large downstream vane, the flow conditions in the rotor passage, at any instant in time, are close to being steady state.