Mechanisms of migraine aura revealed by functional MRI in human visual cortex

Cortical spreading depression (CSD) has been suggested to underlie migraine visual aura. However, it has been challenging to test this hypothesis in human cerebral cortex. Using high-field functional MRI with near-continuous recording during visual aura in three subjects, we observed blood oxygenation level-dependent (BOLD) signal changes that demonstrated at least eight characteristics of CSD, time-locked to percept/onset of the aura. Initially, a focal increase in BOLD signal (possibly reflecting vasodilation), developed within extrastriate cortex (area V3A). This BOLD change progressed contiguously and slowly (3.5 ± 1.1 mm/min) over occipital cortex, congruent with the retinotopy of the visual percept. Following the same retinotopic progression, the BOLD signal then diminished (possibly reflecting vasoconstriction after the initial vasodilation), as did the BOLD response to visual activation. During periods with no visual stimulation, but while the subject was experiencing scintillations, BOLD signal followed the retinotopic progression of the visual percept. These data strongly suggest that an electrophysiological event such as CSD generates the aura in human visual cortex.

Actin-Bundling Protein Isolated from Pollen Tubes of Lily : Biochemical and Immunocytochemical Characterization

A 135-kD actin-bundling protein was purified from pollen tubes of lily (Lilium longiflorum) using its affinity to F-actin. From a crude extract of the pollen tubes, this protein was coprecipitated with exogenously added F-actin and then dissociated from F-actin by treating it with high-ionic-strength solution. The protein was further purified sequentially by chromatography on a hydroxylapatite column, a gel-filtration column, and a diethylaminoethyl-cellulose ion-exchange column. In the present study, this protein is tentatively referred to as P-135-ABP (Plant 135-kD Actin-Bundling Protein). By the elution position from a gel-filtration column, we estimated the native molecular mass of purified P-135-ABP to be 260 kD, indicating that it existed in a dimeric form under physiological conditions. This protein bound to and bundled F-actin prepared from chicken breast muscle in a Ca2+-independent manner. The binding of 135-P-ABP to actin was saturated at an approximate stoichiometry of 26 actin monomers to 1 dimer of P-135-ABP. By transmission electron microscopy of thin sections, we observed cross-bridges between F-actins with a longitudinal periodicity of 31 nm. Immunofluorescence microscopy using rhodamine-phalloidin and antibodies against the 135-kD polypeptide showed that P-135-ABP was colocalized with bundles of actin filaments in lily pollen tubes, leading us to conclude that it is the factor responsible for bundling the filaments.

Three quaternary structures for a single protein.

The structure of a multisubunit protein (immunoglobulin light chain) was solved in three crystal forms, differing only in the solvent of crystallization. The three structures were obtained at high ionic strength and low pH, high ionic strength and high pH, and low ionic strength and neutral pH. The three resulting "snapshots" of possible structures show that their variable-domain interactions differ, reflecting their stabilities under specific solvent conditions. In the three crystal forms, the variable domains had different rotational and translational relationships, whereas no alteration of the constant domains was found. The critical residues involved in the observed effect of the solvent are tryptophans and histidines located between the two variable domains in the dimeric structure. Tryptophan residues are commonly found in interfaces between proteins and their subunits, and histidines have been implicated in pH-dependent conformation changes. The quaternary structure observed for a multisubunit protein or protein complex in a crystal may be influenced by the interactions of the constituents within the molecule or complex and/or by crystal packing interactions. The comparison of buried surface areas and hydrogen bonds between the domains forming the molecule and between the molecules forming the crystals suggest that, for this system, the interactions within the molecule are most likely the determining factors.

Mammalian phospholipase D: activation by ammonium sulfate and nucleotides.

Phospholipase D (PLD) associated with the rat kidney membrane was activated by guanine 5'-[gamma-thio]triphosphate and a cytosol fraction that contained ADP-ribosylation factor. When assayed by measuring the phosphatidyl transfer reaction to ethanol with exogenously added radioactive phosphatidylcholine as substrate, the PLD required a high concentration (1.6 M) of ammonium sulfate to exhibit high enzymatic activity. Other salts examined were far less effective or practically inactive, and this dramatic action of ammonium sulfate is not simply due to such high ionic strength. Addition of ATP but not of nonhydrolyzable ATP analogue adenosine 5'-[beta, gamma-imido]diphosphate further enhanced the PLD activation approximately equal to 2- to 3-fold. This enhancement by ATP needed cytosol, implying a role of protein phosphorylation. A survey of PLD activity in rat tissues revealed that, unlike in previous observations reported thus far, PLD was most abundant in membrane fractions of kidney, spleen, and liver in this order, and the enzymatic activity in brain and lung was low.

Association of a transglutaminase-related antigen with intermediate filaments.

A mouse monoclonal antibody, G92.1.2, raised against guinea pig liver transglutaminase (TGase) recognizes an antigen present in primary mouse dermal fibroblasts. A filamentous pattern, bearing remarkable similarity to the vimentin intermediate filament (IF) network, is seen when these cells are fixed and processed for indirect immunofluorescence with the antibody. Double-label immunofluorescence reveals that the antigen reacting with the antibody colocalizes precisely with vimentin IF and that this colocalization is retained after the treatment of fibroblasts with colchicine, which induces a redistribution of the majority of IFs into perinuclear aggregates. These morphological observations are further supported by the finding that the protein reacting with G92.1.2 is retained in IF-enriched cytoskeletal preparations made by using nonionic detergent-containing high ionic strength solutions. Western blots of the IF fraction show that G92.1.2 recognizes a major band of approximately 280 kDa and does not cross react with vimentin. Furthermore, when the antibody is microinjected into live dermal fibroblasts, it causes a collapse of the vimentin IF network in the majority of injected cells. The results suggest that a form of TGase, or a TGase-related antigen, is closely associated with the vimentin IF network of primary cultures of mouse dermal fibroblasts.

Light-generated nuclear quantum beats: a signature of photosynthesis.

Light-induced radical pairs in deuterated and deuterated plus 15N-substituted Synechococcus lividus cyanobacteria have been studied by transient EPR following pulsed laser excitation. Nuclear quantum beats are observed in the transverse electron magnetization at lower temperatures. Model calculations for the time profiles, evaluated at the high-field emissive maximum of the spectrum, indicate assignment of these coherences to nitrogen nuclei in the primary donor. Thorough investigation of the nuclear modulation patterns can provide detailed information on the electronic structure of the primary donor, providing insight into the mechanism of the primary events of plant photosynthesis.

Trapping of branched DNA in microfabricated structures.

We have observed electrostatic trapping of tribranched DNA molecules undergoing electrophoresis in a microfabricated pseudo-two-dimensional array of posts. Trapping occurs in a unique transport regimen in which the electrophoretic mobility is extremely sensitive to polymer topology. The arrest of branched polymers is explained by considering their center-of-mass motion; in certain conformations, owing to the constraints imposed by the obstacles a molecule cannot advance without the center of mass first moving a short distance backwards. The depth of the resulting local potential well can be much greater than the thermal energy so that escape of an immobilized molecule can be extremely slow. We summarize the expected behavior of the mobility as a function of field strength and topology and point out that the microfabricated arrays are highly suitable for detecting an extremely small number of branched molecules in a very large population of linear molecules.

Aplicação de gelatina obtida de subproduto animal como substituto parcial de gordura em spread de chocolate

O objetivo do presente estudo foi verificação da viabilidade da aplicação de gelatina de pés de frango em spread de chocolate como substituto parcial de gordura. Inicialmente apresentou-se uma revisão de literatura sobre os principais aspectos que envolvem a pesquisa. Após relatou-se a metodologia de extração e caracterização físico-química da gelatina obtida de pés de frango por meio de análises de composição centesimal, de cor, força de gel e análise de perfil de textura (TPA). Posteriormente abordou-se os efeitos da aplicação de gelatina como substituto de gordura nas propriedades físico-químicas e reológicas de spread de chocolate. Por último investigou-se a influência da concentração de gelatina e substituição de gordura na espalhabilidade, estabilidade (shelf life), custos e propriedades sensoriais dos spreads de chocolate. Para tanto, foi utilizado planejamento experimental Central Composto Rotacional e metodologia de superfície de resposta. Os resultados da extração de gelatina de subproduto de frango indicaram que as peles e tendões dos pés de frango possuem rendimento (7,83 %) e conteúdo mineral (1,92 %), apresentando alto teor proteico (84,96 %) próximo das gelatinas comerciais. Em concentração de 6,67 % a gelatina demonstrou elevada força de gel (294,78 g), comportando-se na análise de perfil de textura como um sólido. Os dados relacionados às formulações de spread de chocolate demonstraram que tanto o nível de substituição de gordura quanto a concentração de gelatina exerceram efeitos significativos (p<0,05) sobre a cor, volume, densidade e comportamento reológico dos produtos, exceto a atividade de água que foi influenciada apenas pelo primeiro fator. Sob a perspectiva reológica apenas a amostra com maior nível de substituição de gordura foi classificada como pseudoplástica. A gelatina contribuiu no aumento do teor proteico das formulações. A consistência foi significativamente (p<0,05) influenciada pelas propriedades da fase de gelatina, apresentando ampla faixa de plasticidade nas diferentes temperaturas (10, 20 e 30 °C). A consistência dos spreads aumentou em todas as temperaturas durante os 90 dias de armazenamento. Observou-se por meio da análise sensorial que os fatores em estudo provocaram alterações nos atributos sensoriais do produto suficiente para a percepção dos provadores. A formulação com 75 % de substituição de gordura seguida pelas formulações com 50 % e 0,8 % de concentração de gelatina apresentaram a maior aceitabilidade. Com o máximo de substituição de gordura chegou-se a 53 % de redução de custos. Contudo, os resultados obtidos sugerem que a aplicação de gelatina de pés de frango pode ser vantajosa, já que contribuiu com a formulação de spreads de chocolate menos calóricos e com propriedades físico-químicas e sensoriais adequadas para uma possível comercialização.

Estudo da geometria da aresta de corte de ferramentas aplicadas ao torneamento de superligas à base de níquel com alta velocidade de corte

Pesquisadores e indústrias de todo o mundo estão firmemente comprometidos com o propósito de fazer o processo de usinagem ser precisamente veloz e produtivo. A forte concorrência mundial gerou a procura por processos de usinagem econômicos, com grande capacidade de produção de cavacos e que produzam peças com elevada qualidade. Dentre as novas tecnologias que começaram a ser empregadas, e deve tornar-se o caminho certo a ser trilhado na busca da competitividade em curto espaço de tempo, está a tecnologia de usinagem com altas velocidades (HSM de High Speed Machining). A tecnologia HSM surge como componente essencial na otimização dos esforços para manutenção e aumento da competitividade global das empresas. Durante os últimos anos a usinagem com alta velocidade tem ganhado grande importância, sendo dada uma maior atenção ao desenvolvimento e à disponibilização no mercado de máquinas-ferramentas com rotações muito elevadas (20.000 - 100.000 rpm). O processo de usinagem com alta velocidade está sendo usado não apenas para ligas de alumínio e cobre, mas também para materiais de difícil usinabilidade, como os aços temperados e superligas à base de níquel. Porém, quando se trata de materiais de difícil corte, têm-se observado poucas publicações, principalmente no processo de torneamento. Mas, antes que a tecnologia HSM possa ser empregada de uma forma econômica, todos os componentes envolvidos no processo de usinagem, incluindo a máquina, o eixo-árvore, a ferramenta e o pessoal, precisam estar afinados com as peculiaridades deste novo processo. No que diz respeito às máquinas-ferramenta, isto significa que elas têm que satisfazer requisitos particulares de segurança. As ferramentas, devido à otimização de suas geometrias, substratos e revestimentos, contribuem para o sucesso deste processo. O presente trabalho objetiva estudar o comportamento de diversas geometrias ) de insertos de cerâmica (Al2O3 + SiCw e Al2O3 + TIC) e PCBN com duas concentrações de CBN na forma padrão, assim como modificações na geometria das arestas de corte empregadas em torneamento com alta velocidade em superligas à base de níquel (Inconel 718 e Waspaloy). Os materiais foram tratados termicamente para dureza de 44 e 40 HRC respectivamente, e usinados sob condição de corte a seco e com utilização da técnica de mínima quantidade de lubrificante (minimal quantity lubricant - MQL) visando atender requisitos ambientais. As superligas à base de níquel são conhecidas como materiais de difícil usinabilidade devido à alta dureza, alta resistência mecânica em alta temperatura, afinidade para reagir com materiais da ferramenta e baixa condutividade térmica. A usinagem de superligas afeta negativamente a integridade da peça. Por essa razão, cuidados especiais devem ser tomados para assegurar a vida da ferramenta e a integridade superficial de componentes usinados por intermédio de controle dos principais parâmetros de usinagem. Experimentos foram realizados sob diversas condições de corte e geometrias de ferramentas para avaliação dos parâmetros: força de corte, temperatura, emissão acústica e integridade superficial (rugosidade superficial, tensão residual, microdureza e microestrutura) e mecanismos de desgaste. Mediante os resultados apresentados, recomenda-se à geometria de melhor desempenho nos parâmetros citados e confirma-se a eficiência da técnica MQL. Dentre as ferramentas e geometrias testadas, a que apresentou melhor desempenho foi a ferramenta cerâmica CC650 seguida da ferramenta cerâmica CC670 ambas com formato redondo e geometria 2 (chanfro em T de 0,15 x 15º com raio de aresta de 0,03 mm).

O efeito de diferentes volumes de treinamento de força nas adaptações funcionais e morfológicas da musculatura esquelética em indivíduos treinados

O objetivo desse estudo foi verificar o efeito de diferentes volumes de treinamento de força na força máxima de membros inferiores e na hipertrofia do reto femoral e do vasto lateral após quatro, oito e doze semanas em indivíduos treinados em força. Vinte e seis indivíduos jovens saudáveis do sexo masculino (idade 23,6 ± 4,6 anos, massa corporal 76,6 ± 7,5 kg, estatura 1,75 ± 0,1 cm), com tempo médio de treinamento de força (4,7 ± 4,1 anos) foram divididos em três grupos experimentais, treinamento de força alto volume (TFAV, n = 8), treinamento de força médio volume (TFMV, n = 9) e treinamento de força baixo volume (TFBV, n = 9). As medidas de força dinâmica máxima (1RM) e de área de secção transversa muscular (ASTM) do reto femoral (RF) e do vasto lateral (VL) foram realizadas nos momentos pré- treinamento, pós quatro semanas, pós oito semanas e pós-treinamento. O volume total de treinamento apresentou aumento estatístico para todos os grupos TFAV (p < 0,0001), TFMV (p < 0,0001) e TFBV (p < 0,0001) ao longo do período experimental. Os valores de 1RM aumentaram de maneira significativa após a oitava semana de treinamento TFAV (11,8 ± 4,7%; p < 0,0001) e TFMV (12,1 ± 8,5%; p < 0,0001) e TFBV (9,6 ± 7,3%; p < 0,001) e no pós-treinamento TFAV (13,9 ± 3,9%; p < 0,0001), TFMV (16,7 ± 10,8%; p < 0,0001) e TFBV (14,0 ± 8,1%; p < 0,0001) para todos os grupos, porém não foi observado diferença entre os grupos. A ASTM do RF apresentou aumento estatístico no pós-treinamento somente para o grupo TFAV (15,0 ± 11,9%; p < 0,0001). Apenas o grupo TFAV aumentou estatisticamente a ASTM do VL após quatro semanas de treinamento (7,71 ± 4,42%; p < 0,0001), porém todos os grupos aumentaram significativamente a ASTM do VL após oito semanas de treinamento TFAV (11,37 ± 3,88%; p < 0,0001), TFMV (9,68 ± 9,36%; p < 0,0001) e TFBV (7,26 ± 3,15%; p < 0,01) e no pós-treinamento TFAV (14,54 ± 4,07%; p < 0,0001), TFMV (14,77 ± 8,24%; p < 0,0001) e TFBV (8,66 ± 3,97%; p < 0,001), porém não foi observado diferença entre os grupos. Os resultados do presente estudo demonstraram que, independente do volume adotado, os ganhos de força máxima foram semelhantes. Por outro lado, a ASTM foi influenciada pelo volume de treinamento, dado que o grupo TFAV foi o único que apresentou aumento significativo da ASTM do RF no pós-treinamento e aumentou a ASTM do VL com apenas quatro semanas de treinamento

A new code for the Hall-driven magnetic evolution of neutron stars

Over the past decade, the numerical modeling of the magnetic field evolution in astrophysical scenarios has become an increasingly important field. In the crystallized crust of neutron stars the evolution of the magnetic field is governed by the Hall induction equation. In this equation the relative contribution of the two terms (Hall term and Ohmic dissipation) varies depending on the local conditions of temperature and magnetic field strength. This results in the transition from the purely parabolic character of the equations to the hyperbolic regime as the magnetic Reynolds number increases, which presents severe numerical problems. Up to now, most attempts to study this problem were based on spectral methods, but they failed in representing the transition to large magnetic Reynolds numbers. We present a new code based on upwind finite differences techniques that can handle situations with arbitrary low magnetic diffusivity and it is suitable for studying the formation of sharp current sheets during the evolution. The code is thoroughly tested in different limits and used to illustrate the evolution of the crustal magnetic field in a neutron star in some representative cases. Our code, coupled to cooling codes, can be used to perform long-term simulations of the magneto-thermal evolution of neutron stars.

Heterofunctional Supports in Enzyme Immobilization: From Traditional Immobilization Protocols to Opportunities in Tuning Enzyme Properties

A heterofunctional support for enzyme immobilization may be defined as that which possesses several distinct functionalities on its surface able to interact with a protein. We will focus on those supports in which a final covalent attachment between the enzyme and the support is achieved. Heterofunctionality sometimes has been featured in very old immobilization techniques, even though in many instances it has been overlooked, giving rise to some misunderstandings. In this respect, glutaraldehyde-activated supports are the oldest multifunctional supports. Their matrix has primary amino groups, the hydrophobic glutaraldehyde chain, and can covalently react with the primary amino groups of the enzyme. Thus, immobilization may start (first event of the immobilization) via different causes and may involve different positions of the enzyme surface depending on the activation degree and immobilization conditions. Other “classical” heterofunctional supports are epoxy commercial supports consisting of reactive covalent epoxy groups on a hydrophobic matrix. Immobilization is performed at high ionic strength to permit protein adsorption, so that covalent attachment may take place at a later stage. Starting from these old immobilization techniques, tailor-made heterofunctional supports have been designed to permit a stricter control of the enzyme immobilization process. The requirement is to find conditions where the main covalent reactive moieties may have very low reactivity toward the enzyme. In this Review we will discuss the suitable properties of the groups able to give the covalent attachment (intending a multipoint covalent attachment), and the groups able to produce the first enzyme adsorption on the support. Prospects, limitations, and likely pathways for the evolution (e.g., coupling of site-directed mutagenesis and thiol heterofunctional supports of enzyme immobilization on heterofunctional supports) will be discussed in this Review.

Comparison Between Dcrit Considering the Abrupt Variation and Inflexion in the Concrete Mercury Intrusion Porosimetry Curve

Mercury intrusion porosimetry (MIP) has been widely used to evaluate the quality of concrete through the pore size distribution parameters. Two of these parameters are the critical pore diameter (Dcrit) and the percentage of the most interconnected net of pores compared to the total volume of pores. Some researchers consider Dcrit as the diameter obtained from the inflexion point of the cumulative mercury intrusion curve while others consider Dcrit as the diameter obtained from the point of abrupt variation in the same curve. This study aims to analyze two groups of concretes of varying w/c ratios, one cast with pozzolanic cement and another with high initial strength cement, in order to determine which of these diameters feature a better correlation with the quality parameters of the concretes. The concrete quality parameters used for the evaluations were (1) the w/c ratios and (2) chloride diffusion coefficients measured at approximately 90 days. MIP cumulative distributions of the same concretes were also measured at about 90 days, and Dcrit values were determined (1) from the point of abrupt variation and alternatively, (2) from the inflexion point of each of these plots. It was found that Dcrit values measured from the point of abrupt variation were useful indicators of the quality of the concrete, but the Dcrit values based on the inflexion points were not. Hence, it is recommended that Dcrit and the percentage of the most interconnected volume of pores should be obtained considering the point of abrupt variation of the cumulative curve of pore size distribution.

The discovery, monitoring and environment of SGR J1935+2154

We report on the discovery of a new member of the magnetar class, SGR J1935+2154, and on its timing and spectral properties measured by an extensive observational campaign carried out between 2014 July and 2015 March with Chandra and XMM–Newton (11 pointings). We discovered the spin period of SGR J1935+2154 through the detection of coherent pulsations at a period of about 3.24 s. The magnetar is slowing down at a rate of P˙=1.43(1)×10−11 s s−1 and with a decreasing trend due to a negative P¨ of −3.5(7) × 10−19 s s−2. This implies a surface dipolar magnetic field strength of ∼2.2 × 1014 G, a characteristic age of about 3.6 kyr and a spin-down luminosity Lsd ∼1.7 × 1034 erg s−1. The source spectrum is well modelled by a blackbody with temperature of about 500 eV plus a power-law component with photon index of about 2. The source showed a moderate long-term variability, with a flux decay of about 25 per cent during the first four months since its discovery, and a re-brightening of the same amount during the second four months. The X-ray data were also used to study the source environment. In particular, we discovered a diffuse emission extending on spatial scales from about 1 arcsec up to at least 1 arcmin around SGR J1935+2154 both in Chandra and XMM–Newton data. This component is constant in flux (at least within uncertainties) and its spectrum is well modelled by a power-law spectrum steeper than that of the pulsar. Though a scattering halo origin seems to be more probable we cannot exclude that part, or all, of the diffuse emission is due to a pulsar wind nebula.

Magnetic anisotropy reveals Neogene tectonic overprint in highly strained carbonate mylonites from the Morcles nappe, Switzerland

The anisotropy of magnetic susceptibility (AMS) has been measured with low- and high-field methods, in deformed carbonate rocks along the Morcles nappe shear zone (Helvetic Alps). High-field measurements at room temperature and 77 K enable the separation of the ferrimagnetic, paramagnetic and diamagnetic anisotropy. The ferrimagnetic sub-fabric is generally insignificant in these rocks, contributing less than 10% to the total AMS. AMS results for both the separated diamagnetic and paramagnetic subfabrics are consistent with the regional shear movement in the late-stage formation of the Helvetic nappes, as seen in the Morcles nappe, whose inverted limb indicate shear displacement towards the northwest. The diamagnetic anisotropy correlates well quantitatively with the calculated magnetic anisotropy based on the calcite texture. There is a gradational change in the degree of anisotropy related to the strain gradient along the shear zone. A more complex magnetic fabric, resulting from partial overprinting due to displacement along the Simplon–Rhône fault, is evident at one site near the root zone of the nappe. Partial overprinting of the magnetic fabric appears to have taken place in two locations farther up the shear zone as well. This late phase deformation is associated with recent exhumation of the Mont Blanc and Belledonne external massifs and orogen parallel extension, and is reflected by the AMS. Rocks with bulk susceptibility ∼0 SI, and simple mineral compositions are ideal for low temperature high-field torque, as this method helps to enhance the paramagnetic susceptibility and anisotropy, which may otherwise be masked by the mixed magnetic contributions of the composite magnetic fabric.