Nerve growth factor induces neurite outgrowth of PC12 cells by promoting Gβγ-microtubule interaction

Background: Assembly and disassembly of microtubules (MTs) is critical for neurite outgrowth and differentiation. Evidence suggests that nerve growth factor (NGF) induces neurite outgrowth from PC12 cells by activating the receptor tyrosine kinase, TrkA. G protein-coupled receptors (GPCRs) as well as heterotrimeric G proteins are also involved in regulating neurite outgrowth. However, the possible connection between these pathways and how they might ultimately converge to regulate the assembly and organization of MTs during neurite outgrowth is not well understood. Results: Here, we report that Gβγ, an important component of the GPCR pathway, is critical for NGF-induced neuronal differentiation of PC12 cells. We have found that NGF promoted the interaction of Gβγ with MTs and stimulated MT assembly. While Gβγ-sequestering peptide GRK2i inhibited neurite formation, disrupted MTs, and induced neurite damage, the Gβγ activator mSIRK stimulated neurite outgrowth, which indicates the involvement of Gβγ in this process. Because we have shown earlier that prenylation and subsequent methylation/demethylation of γ subunits are required for the Gβγ-MTs interaction in vitro, small-molecule inhibitors (L-28 and L-23) targeting prenylated methylated protein methyl esterase (PMPMEase) were tested in the current study. We found that these inhibitors disrupted Gβγ and ΜΤ organization and affected cellular morphology and neurite outgrowth. In further support of a role of Gβγ-MT interaction in neuronal differentiation, it was observed that overexpression of Gβγ in PC12 cells induced neurite outgrowth in the absence of added NGF. Moreover, overexpressed Gβγ exhibited a pattern of association with MTs similar to that observed in NGF-differentiated cells. Conclusions: Altogether, our results demonstrate that βγ subunit of heterotrimeric G proteins play a critical role in neurite outgrowth and differentiation by interacting with MTs and modulating MT rearrangement. Electronic supplementary material The online version of this article (doi:10.1186/s12868-014-0132-4) contains supplementary material, which is available to authorized users

Tomography of H-alpha emission in TCABR Tokamak

Neste trabalho foi feito um estudo do perfil tomográfico da emissão da linha do átomo de hidrogênio, H-alfa (?=656,28 nm) no plasma do TCABR, um tokamak de porte médio em operação no Laboratório de Física de Plasmas do Instituto de Física da Universidade de São Paulo. Nosso trabalho centrou-se no estudo dos efeitos da introdução de um eletrodo polarizado na borda do plasma no tokamak TCABR. O eletrodo pode ser introduzido até 1,5 cm para dentro da coluna do plasma, sem causar disrupturas para polarização positiva de 0 até +350V, e situado no plano equatorial do tokamak. Perfis tomográficos de H-alfa com e sem polarização foram medidos. A comparação dos perfis mostra um aumento da densidade de linha na posição central, quando a emissividade H-alfa diminui. A análise dos perfis tomográficos de H-alfa, tempo de confinamento das partículas e também do estudo de reciclagem das partículas neutras, indica que o plasma entra no regime de alto confinamento (modo-H). Cálculos de turbulência e de transporte na borda do plasma (SOL), feitos medindo o potencial flutuante e a corrente de saturação de íons, mostram uma diminuição forte no espectro de potência e de transporte. Também foram feitos estudos do novo regime de descargas com elétrons fugitivos (\"runaway electron\"), descoberto no tokamak TCABR. As características distintivas deste regime são um plasma de baixa temperatura fracamente ionizado, destacado do limitador devido a processos de recombinação, e instabilidade de relaxação com fortes picos de emissão H-alfa correlacionados com instabilidade dente de serra da densidade eletrônica de linha. No presente trabalho fazemos a descrição das condições experimentais para a geração destas descargas. A produção dos elétrons fugitivos é analisada; mostrando que a geração de elétrons fugitivos somente pode ser explicada pelo mecanismo de avalanche. A confirmação de baixa temperatura do plasma é obtida de uma análise do perfil tomográfico da emissão H-alfa. Esta emissão não pode ser explicada por excitação de elétrons no plasma. A recombinação, de outro lado, dá uma explicação plausível para a dependência temporal da emissão, em particular para alta densidade de partículas neutras.A study of the tomography profile of the emission of the line of Hydrogen, atomic H-alpha line (?=656.28 nm), was carried out in TCABR, a medium-size tokamak in operation at the Laboratory of Plasma Physics of the Institute of Physics of the University of São Paulo. Our work focuses on the study of the effects of due to the introduction of a biased electrode in the plasma edge of the TCABR tokamak. The electrode could be introduced up to 1.5 cm inside the plasma, without plasma disruptions for positive voltages from 0 to +350V, and was located on the equatorial plane of the plasma column. Tomography profiles of H-alpha with and without bias were measured. Comparison of the profiles shows an increase of the central line-averaged density, while the emissivity of the line H-alpha decreases. The analysis of the tomography profiles of H-alpha, time of confinement of particles and also the study of recycling of the neutral particles, indicate that the confined plasma enters the H-mode regime. Calculations of turbulence and transport at the Scrape-Off-Layer, using measured floating potentials and ion saturation currents, show a strong decrease in the power spectra and transport. The H-alpha tomography was also employed to study the new regime of runaway discharges that has been discovered in the TCABR tokamak. The distinctive features of this regime are weakly ionized low-temperature plasma detached from the limiter due to the recombination process, and a relaxation instability with strong spikes of H-alpha emission correlated with sawtooth relaxation of the line density. In the present thesis we report experimental data on conditions for generation of these discharges. The runaway electron production is analyzed; show that generation of runaway electrons can only be explained by the runaway avalanche mechanism. The confirmation of low plasma temperature is a obtained from an analysis of the tomography profile of H-alpha emission. This emission cannot be explained by excitation by plasma electrons. Recombination, on the other hand, gives a rather plausible explanation for the time dependency of the emission, in particular at high neutral densities

Tomography of H-alpha emission in TCABR Tokamak

Neste trabalho foi feito um estudo do perfil tomográfico da emissão da linha do átomo de hidrogênio, H-alfa (?=656,28 nm) no plasma do TCABR, um tokamak de porte médio em operação no Laboratório de Física de Plasmas do Instituto de Física da Universidade de São Paulo. Nosso trabalho centrou-se no estudo dos efeitos da introdução de um eletrodo polarizado na borda do plasma no tokamak TCABR. O eletrodo pode ser introduzido até 1,5 cm para dentro da coluna do plasma, sem causar disrupturas para polarização positiva de 0 até +350V, e situado no plano equatorial do tokamak. Perfis tomográficos de H-alfa com e sem polarização foram medidos. A comparação dos perfis mostra um aumento da densidade de linha na posição central, quando a emissividade H-alfa diminui. A análise dos perfis tomográficos de H-alfa, tempo de confinamento das partículas e também do estudo de reciclagem das partículas neutras, indica que o plasma entra no regime de alto confinamento (modo-H). Cálculos de turbulência e de transporte na borda do plasma (SOL), feitos medindo o potencial flutuante e a corrente de saturação de íons, mostram uma diminuição forte no espectro de potência e de transporte. Também foram feitos estudos do novo regime de descargas com elétrons fugitivos (\"runaway electron\"), descoberto no tokamak TCABR. As características distintivas deste regime são um plasma de baixa temperatura fracamente ionizado, destacado do limitador devido a processos de recombinação, e instabilidade de relaxação com fortes picos de emissão H-alfa correlacionados com instabilidade dente de serra da densidade eletrônica de linha. No presente trabalho fazemos a descrição das condições experimentais para a geração destas descargas. A produção dos elétrons fugitivos é analisada; mostrando que a geração de elétrons fugitivos somente pode ser explicada pelo mecanismo de avalanche. A confirmação de baixa temperatura do plasma é obtida de uma análise do perfil tomográfico da emissão H-alfa. Esta emissão não pode ser explicada por excitação de elétrons no plasma. A recombinação, de outro lado, dá uma explicação plausível para a dependência temporal da emissão, em particular para alta densidade de partículas neutras.A study of the tomography profile of the emission of the line of Hydrogen, atomic H-alpha line (?=656.28 nm), was carried out in TCABR, a medium-size tokamak in operation at the Laboratory of Plasma Physics of the Institute of Physics of the University of São Paulo. Our work focuses on the study of the effects of due to the introduction of a biased electrode in the plasma edge of the TCABR tokamak. The electrode could be introduced up to 1.5 cm inside the plasma, without plasma disruptions for positive voltages from 0 to +350V, and was located on the equatorial plane of the plasma column. Tomography profiles of H-alpha with and without bias were measured. Comparison of the profiles shows an increase of the central line-averaged density, while the emissivity of the line H-alpha decreases. The analysis of the tomography profiles of H-alpha, time of confinement of particles and also the study of recycling of the neutral particles, indicate that the confined plasma enters the H-mode regime. Calculations of turbulence and transport at the Scrape-Off-Layer, using measured floating potentials and ion saturation currents, show a strong decrease in the power spectra and transport. The H-alpha tomography was also employed to study the new regime of runaway discharges that has been discovered in the TCABR tokamak. The distinctive features of this regime are weakly ionized low-temperature plasma detached from the limiter due to the recombination process, and a relaxation instability with strong spikes of H-alpha emission correlated with sawtooth relaxation of the line density. In the present thesis we report experimental data on conditions for generation of these discharges. The runaway electron production is analyzed; show that generation of runaway electrons can only be explained by the runaway avalanche mechanism. The confirmation of low plasma temperature is a obtained from an analysis of the tomography profile of H-alpha emission. This emission cannot be explained by excitation by plasma electrons. Recombination, on the other hand, gives a rather plausible explanation for the time dependency of the emission, in particular at high neutral densities

Oxitec and MosquitoMate in the United States: lessons for the future of gene drive mosquito control.

In response to growing concerns regarding mosquito-borne diseases, scientists are developing novel systems of vector control. Early examples include Oxitec's OX513A genetically-engineered mosquito and MosquitoMate's Wolbachia-infected mosquito, and systems using 'gene-drive' are in development. Systems based on genetic engineering are controversial and institutions around the world are grappling with the question of who should have a say in how such technologies are field-tested and used. Based on media coverage and public records, we created comparative timelines of the efforts of Oxitec and MosquitoMate to navigate federal and local governance and bring their products to market in the United States. We analyze these timelines with particular attention to the role of public input in technology governance. These cases illustrate how governance of technology in the US is diverse, complex, and opaque. Further, the public response to proposed field trials of the Oxitec product highlights inconsistencies between public expectations for governance and actual practice. As gene-drive mosquito control products develop, both federal and local agencies will find their legitimacy tested without a better procedure for transparently integrating public input

Non spirographic or noninvasive methods to estimate anaerobic treshold

In the world of sports research, there are different ways to determinate physical conditioning, ranging from expensive laboratory-invasive methods to cheap, field based-non-invasive methods. The field based-non-invasive test maintains good reliability and low cost using physiological parameters such heart rate, saliva electrolytes or lactate, perceived exertion and electromyography among others. These parameters can be used to estimate anaerobic threshold (AnT) to predict sport performance, redirect training and can help coaches and athletes to be more competitive. However, each of this parameter has some particularities and controversy due to different results reported by specialist. These differences may be explained by protocol testing used, sport level sample, starting intensity or number of levels among others. Despite this, they still have good reproducibility and applications on field based test protocols. Mentioned tests could be used in a large scale, and more often, with paying attention about level of correlation with original invasive tests, and percent of possible mistake in estimation process. Cheaper, and simpler tests (instead subjective estimation of training load) allows using more precise planning and changing volume, and intensity of training, for coaches, and athletes, that have needed level of education and less money. Athletes comfort, and possible high frequency of testing during non - invasive test, also must be emphasized as an advantage during training evaluation process

Comparative electron temperature measurements of Thomson scattering and electron cyclotron emission diagnostics in TCABR plasmas

We present the first simultaneous measurements of the Thomson scattering and electron cyclotron emission radiometer diagnostics performed at TCABR tokamak with Alfven wave heating. The Thomson scattering diagnostic is an upgraded version of the one previously installed at the ISTTOK tokamak, while the electron cyclotron emission radiometer employs a heterodyne sweeping radiometer. For purely Ohmic discharges, the electron temperature measurements from both diagnostics are in good agreement. Additional Alfven wave heating does not affect the capability of the Thomson scattering diagnostic to measure the instantaneous electron temperature, whereas measurements from the electron cyclotron emission radiometer become underestimates of the actual temperature values. (C) 2010 American Institute of Physics. [doi:10.1063/1.3494379

Temporal behaviour of toroidal rotation velocity in the TCABR tokamak

A new method for determining the temporal evolution of plasma rotation is reported in this work. The method is based upon the detection of two different portions of the spectral profile of a plasma impurity line, using a monochromator with two photomultipliers installed at the exit slits. The plasma rotation velocity is determined by the ratio of the two detected signals. The measured toroidal rotation velocities of C III (4647.4 angstrom) and C VI (5290.6 angstrom), at different radial positions in TCABR discharges, show good agreement, within experimental uncertainty, with previous results (Severo et al 2003 Nucl. Fusion 43 1047). In particular, they confirm that the plasma core rotates in the direction opposite to the plasma current, while near the plasma edge (r/a > 0.9) the rotation is in the same direction. This technique was also used to investigate the dependence of toroidal rotation on the poloidal position of gas puffing. The results show that there is no dependence for the plasma core, while for plasma edge (r/a > 0.9) some dependence is observed.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Foundation for Supporting Research of the State of Sao Paulo (FAPESP)National Fusion Network (RNF/FINEP)Financiadora de Estudos e Projetos (FINEP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)National Councel of Research and Technological Development (CNPq

Long-distance correlations in TCABR biasing experiments

Long-distance correlations (LDCs) of plasma potential fluctuations in the plasma edge have been investigated in the TCABR tokamak in the regime of edge biasing H-mode using an array of multi-pin Langmuir probes. This activity was carried out as part of the scientific programme of the 4th IAEA Joint Experiment (2009). The experimental data confirm the effect of amplification of LDCs in potential fluctuations during biasing recently observed in stellarators and tokamaks. For long toroidal distances between probes, the cross-spectrum is concentrated at low frequencies f < 60 kHz with peaks at f < 5 kHz, f = 13-15 kHz and f similar to 40 kHz and low wave numbers with a maximum at k = 0. The effects of MHD activity on the LDCs in potential fluctuation are investigated.FINEPFINEPFAPESPFAPESPIAEA (International Atomic Energy Agency)IAEA (International Atomic Energy Agency)National Fusion NetworkNational Fusion Networ