Indicadores de cálcio e de voltagem codificados geneticamente na detecção de potenciais de ação e inputs sinápticos em cultura de neurônios hipocampais

Recently, genetically encoded optical indicators have emerged as noninvasive tools of high spatial and temporal resolution utilized to monitor the activity of individual neurons and specific neuronal populations. The increasing number of new optogenetic indicators, together with the absence of comparisons under identical conditions, has generated difficulty in choosing the most appropriate protein, depending on the experimental design. Therefore, the purpose of our study was to compare three recently developed reporter proteins: the calcium indicators GCaMP3 and R-GECO1, and the voltage indicator VSFP butterfly1.2. These probes were expressed in hippocampal neurons in culture, which were subjected to patchclamp recordings and optical imaging. The three groups (each one expressing a protein) exhibited similar values of membrane potential (in mV, GCaMP3: -56 ±8.0, R-GECO1: -57 ±2.5; VSFP: -60 ±3.9, p = 0.86); however, the group of neurons expressing VSFP showed a lower average of input resistance than the other groups (in Mohms, GCaMP3: 161 ±18.3; GECO1-R: 128 ±15.3; VSFP: 94 ±14.0, p = 0.02). Each neuron was submitted to current injections at different frequencies (10 Hz, 5 Hz, 3 Hz, 1.5 Hz, and 0.7 Hz) and their fluorescence responses were recorded in time. In our study, only 26.7% (4/15) of the neurons expressing VSFP showed detectable fluorescence signal in response to action potentials (APs). The average signal-to-noise ratio (SNR) obtained in response to five spikes (at 10 Hz) was small (1.3 ± 0.21), however the rapid kinetics of the VSFP allowed discrimination of APs as individual peaks, with detection of 53% of the evoked APs. Frequencies below 5 Hz and subthreshold signals were undetectable due to high noise. On the other hand, calcium indicators showed the greatest change in fluorescence following the same protocol (five APs at 10 Hz). Among the GCaMP3 expressing neurons, 80% (8/10) exhibited signal, with an average SNR value of 21 ±6.69 (soma), while for the R-GECO1 neurons, 50% (2/4) of the neurons had signal, with a mean SNR value of 52 ±19.7 (soma). For protocols at 10 Hz, 54% of the evoked APs were detected with GCaMP3 and 85% with R-GECO1. APs were detectable in all the analyzed frequencies and fluorescence signals were detected from subthreshold depolarizations as well. Because GCaMP3 is the most likely to yield fluorescence signal and with high SNR, some experiments were performed only with this probe. We demonstrate that GCaMP3 is effective in detecting synaptic inputs (involving Ca2+ influx), with high spatial and temporal resolution. Differences were also observed between the SNR values resulting from evoked APs, compared to spontaneous APs. In recordings of groups of cells, GCaMP3 showed clear discrimination between activated and silent cells, and reveals itself as a potential tool in studies of neuronal synchronization. Thus, our results indicate that the presently available calcium indicators allow detailed studies on neuronal communication, ranging from individual dendritic spines to the investigation of events of synchrony in neuronal networks genetically defined. In contrast, studies employing VSFPs represent a promising technology for monitoring neural activity and, although still to be improved, they may become more appropriate than calcium indicators, since neurons work on a time scale faster than events of calcium may foresee

Avaliação da resposta comportamental, morfofisiológica e produção de leite de cabras puras e mestiças da raça Saanen no semi-árido do Rio Grande do Norte

Goat breeding in the state of Rio Grande do Norte, Brazil has promising economic possibilities, with the proper handling of the natural resources. The introduction of specialized animals has been one of the ways used to improve herd genetics and increase productivity. However, climate has been one of the regional factors that most interferes with the adaptation of the new genetic prevalence resulting from the introduction of exotic breeds, because in their country of origin, the air temperature during most of the year is lower than the animals body temperature. With this in mind, the aim of this study was to characterize behavioral, physiological and morphological profiles and milk production of female Saanen goats belonging to different genetic groups raised in the semi-arid region of Rio Grande do Norte in Northeast Brazil. The study was conducted in the city of Lages (5° 42 00 S and 36° 14 41 W). We used 25 lactating female Saanen goats, distributed into 3 genetic groups: 5 purebred animals, 11 three-quarter bred and 9 half-bred. Behavioral observations were made over three consecutive days in the months of August and September, between 09:00 and 11:30h, when the animals were grazing. Physiological and meteorological data were recorded in the last three days of June, July, August and September at 05:00h and at 16:00h. In the semi-intensive breeding system, the animals from different genetic groups were similar in both field behavior and physiological response patterns. Although the purebred goats had longer hair, they did not show symptoms of thermal discomfort. Their white hair helped to reflect the short wavelength rays and thus eliminate those at the longer wave lengths. We concluded that the animals raised in the semi-intensive milk production system in this study seem to have adapted to the climatic conditions of the semi-arid region of Rio Grande do Norte, Brazil

Expressão de zif-268 durante a aquisição, evocação e extinção de uma memória aversiva

In the behavioral paradigm of discriminative avoidance task, both short and long-term memories have been extensively investigated with behavioral and pharmacological approaches. The aim of the present study was to evaluate, using the abovementioned model, the hippocampal expression of zif-268 - a calcium-dependent immediate early gene involved with synaptic plasticity process - throughout several steps of memory formation, such as acquisition, evocation and extiction. The behavioral apparatus consisted of a modified elevaated plus-maze, with their enclosed arms disposed in "L". A pre-exposure to the maze was made with the animal using all arms enclosed, for 30 minutes, followed by training and test, during 10 minutes each. The between sections interval was 24h. During training, aversive stimuli (bright light and loud noise) were actived whenever the animals entered one of the enclosed armas (aversive arm). Memory acquisiton, retention and extinction were evaluated by the percentage of the total time spent exploring the aversive arm. The parameters evaluated (time spent in the arms and total distance traveled) were estimated with an animal tracking software (Anymaze, Stoelting, USA). Learning during training was estimated by the decrease of the time spent exploring the aversive arm. One hour after the beginning of each section, animals were anaesthetized with sodium-thiopental (i.p.) and perfused with 0.9% heparinized saline solution followed by 4% paraformaldehyde. Brains were cryoprotected with 20% sucrose, separeted in three blocks and frozen. The middle block, containing the hippocampus, was sectioned at 20 micro meters in the coronal plane and the resutant sections were submitted to zif-268 immunohistochemistry. Our results show an increased expression of zif-268 in the dentate gyrus (DG) during the evocation and extinction stages. There is a distinct participation of the DG during the memory evocation, but not during its acquisition. Inaddition, all hippocampal regions (CA1, CA3 and DG) presented an increased zif-268 expression during the process of extinction.

Produtividade fabril I: método para rápido aumento da produtividade fabril

O método para rápido aumento da produtividade fabril, aplicável principalmente à indústria brasileira de manufatura (17% do PIB), fundamenta-se exclusivamente na redução ou eliminação do tempo inativo do homem, da máquina e do material, que é a grande causa da ineficiência. O método exige a utilização de apenas cinco das mais simples, elementares e conhecidas técnicas e é aplicável às vinte situações mais freqüentes (objetos de estudo) na indústria de manufatura. Nossa experiência atesta que o método é capaz de aumentar a produtividade fabril em valor superior a 30%, em poucos meses e de forma perene (houve um caso de aumento de 160% em toda uma seção de usinagem de uma empresa multinacional). Sua aplicação e a implantação das medidas dele decorrentes são feitas com extrema facilidade e, por isto, os resultados surgem muito rapidamente. Atinge portanto seu objetivo: proporcionar aumento da produtividade fabril em curto espaço de tempo. Os demais artigos da série detalharão os objetos de estudo.

Produtividade fabril II - método para rápido aumento da produtividade fabril: redução de esperas dentro do ciclo da operação

O método para rápido aumento da produtividade fabril, aplicável principalmente à indústria brasileira de manufatura (17% do PIB), fundamenta-se exclusivamente na redução ou eliminação do tempo inativo do homem, da máquina e do material, que é a grande causa da ineficiência. O método exige a utilização de apenas cinco das mais simples, elementares e conhecidas técnicas e é aplicável às vinte situações mais freqüentes (objetos de estudo) na indústria de manufatura. Nossa experiência atesta que o método é capaz de aumentar a produtividade fabril em valor superior a 30%, em poucos meses e de forma perene (houve um caso de aumento de 160% em toda uma seção de usinagem de uma empresa multinacional). Sua aplicação e a implantação das medidas dele decorrentes são feitas com extrema facilidade e, por isto, os resultados surgem muito rapidamente. Atinge portanto seu objetivo: proporcionar aumento da produtividade fabril em curto espaço de tempo. Este artigo detalhará sete objetos de estudo (situações): 1.redução da espera da máquina durante as inspeções; 2.redução da espera do operário, abordando trabalho em equipe, produção em linha e operação com ajudante; e 3.redução da espera do operador de máquina, por meio da diminuição do tempo-máquina, do controle do tempo-máquina e da atribuição de outras atividades ao operador pela adoção da célula de manufatura.

Produtividade fabril III - método para rápido aumento da produtividade fabril: redução de tempos inativos e do tempo de espera do material em processo

O método para rápido aumento da produtividade fabril, aplicável principalmente à indústria brasileira de manufatura (17% do PIB), fundamenta-se exclusivamente na redução ou eliminação do tempo inativo do homem, da máquina e do material, que é a grande causa da ineficiência. O método exige a utilização de apenas cinco das mais simples, elementares e conhecidas técnicas e é aplicável às vinte situações mais freqüentes (objetos de estudo) na indústria de manufatura. Nossa experiência atesta que o método pode proporcionar à produtividade fabril um aumento superior a 30%, em poucos meses e de forma perene. Sua aplicação e a implantação das medidas dele decorrentes são feitas com extrema facilidade e, por isto, os resultados surgem muito rapidamente. Atinge portanto seu objetivo: proporcionar aumento da produtividade fabril em curto espaço de tempo. Este artigo (terceiro e último da série) detalhará doze objetos de estudo (situações): 1. redução dos tempos inativos causados por troca de turno, por refeição e por troca de produtos; 2. redução dos tempos improdutivos acarretados por causas mais importantes, por espera pelo serviço de manutenção, por espera pelo serviço de preparação, por espera pelo operador e por causas não apontáveis pelos procedimentos usuais; 3. redução das atividades improdutivas e das produtivas executadas num ritmo improdutivo; e 4. redução do tempo de espera do material em processamento pela redução do tamanho do lote de fabricação e pelo aumento da velocidade de manufatura.

Metodologia para rastreamento com modificação dos zeros e rejeição de distúrbio aplicada a sistemas incertos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Síntese e caracterização de materiais nanoporosos para pirólise catalítica de óleos pesados

The present work reports the study of nanoporous structures, aiming at their use in research directed to the current demand of the petroleum industry to value heavy oil. Initially, two ways were chosen for the synthesis of porous structures from the molecular sieves of type Si-MCM-41. In the first way, the structure MCM-41 is precursory for heteroatom substitutes of silicon, generating catalyst of the type Al-MCM-41 from two different methods of incorporation of the metal. This variation of the incorporation method of Aluminum in the structure of Si-MCM-41 was carried out through the conventional procedure, where the aluminum source was incorporated to the gel of synthesis, and the procedure post-synthesis, where the Aluminum source was incorporated in catalyst after the synthesis of Si-MCM-41. In the second way, the MCM-41 acts as a support for growth of nanocrystals of zeolite embedded in their mesoporous, resulting in hybrid MCM-41/ZSM-5 catalyst. A comparative analysis was carried through characterizations by XRD, FTIR, measures of acidity through n-butylamine adsorption for TGA, SEM-XRF and N2 adsorption. Also crystalline aluminosilicate with zeolitic structure MFI of type ZSM-5 was synthesized without using organic templates. Methodologies to the preparation of these materials are related by literature using conventionally reactants that supply oxides of necessary silicon and aluminum, as well as a template agent, and in some cases co-template. The search for new routes of preparation for the ZSM-5 aimed at, above all, the optimization of the same as for the time and the temperature of synthesis, and mainly the elimination of the use of organic templates, that are material of high cost and generally very toxic. The current study is based on the use of the H2O and Na+ cations playing the role of structural template and charge compensation in the structure. Characterizations by XRD, FTIR, SEM-XRF and N2 adsorption were also conducted for this material in order to compare the samples of ZSM-5 synthesized in the absence of template and those used industrially and synthesized using structuring

Aplicação da tecnologia eletroquímica como alternativa para a remoção de corante têxteis em efluentes sintéticos e reais utilizando anodos de platina e diamante

In this work, electrochemical technology was used to treat synthetic wastewater containing Methyl Red (MR) and Blue Novacron (BN) by anodic oxidation using anodes platinum (Pt) and real samples of textile effluents using DDB anodes and platinum (Pt). The removal of color from the galvanostatic electrolysis of synthetic wastewater MR and BN, and the actual sample has been observed under different conditions (different current densities and temperature variation). The investigation of these parameters was performed in order to establish the best conditions for removal of color and chemical oxygen demand (BOD). According to the results obtained in this study, the electrochemical oxidation processes suitable for the degradation process of color and COD in wastewater containing such textile dyes, because the electrocatalytic properties of Pt and BDD anodes consumption energy during the electrochemical oxidation of synthetic solutions AN and MR and real sample, mainly depend on the operating parameters of operation, for example, the synthetic sample of MR, energy consumption rose from 42,00kWhm-3 in 40 mAcm-2 and 25 C to 17,50 kWhm-3 in 40mAcm-2 and 40 C, from the BN went 17,83 kWhm-3 in 40mAcm and 40°C to 14,04 kWhm- 3 in 40mAcm-2 and 40 C (data estimated by the volume of treated effluent). These results clearly indicate the applicability of electrochemical treatment for removing dyes from synthetic solutions and real industrial effluents

Determinação do teor de cálcio através da termogravimetria, em medicamentos utilizados no tratamento da osteoporose: um estudo comparativo

The present study utilized the thermogravimetry (TG) and optical emission spectroscopy with inductively coupled plasma - ICP / OES to determine the calcium content in tablets of carbonate, citrate and calcium lactate used in the treatment of osteoporosis. The samples were characterized by IR, SEM, TG / DTG, DTA, DSC and XRD. The thermal analysis evaluated the thermal stability and physical-chemical events and showed that the excipients influence the decomposition of active ingredients. The results of thermogravimetry indicated that the decomposition temperature of the active CaCO3 (T = 630.2 °C) is lower compared to that obtained in samples of the tablets (633.4 to 655.2 °C) except for sample AM 2 (Ti = 613.8 oC). In 500.0 °C in the samples of citrate and calcium lactate, as well as their respective active principles had already been formed calcium carbonate. The use of N2 atmosphere resulted in shifting the initial and final temperature related to the decomposition of CaCO3. In the DTA and DSC curves were observed endo and exothermic events for the samples of tablets and active ingredients studied. The infrared spectra identified the main functional groups in all samples of active ingredients, excipients and tablets studied, such as symmetric and asymmetric stretches of the groups OH, CH, C = O. Analysis by X-ray diffraction showed that all samples are crystalline and that the final residue showed peaks indicative of the presence of calcium hydroxide by the reaction of calcium oxide with moisture of the air. Although the samples AM 1, AM 2, AM 3 and AM 6 in their formulations have TiO2 and SiO2 peaks were not observed in X-ray diffractograms of these compounds. The results obtained by TGA to determine the calcium content of the drugs studied were satisfactory when compared with those obtained by ICP-OES. In the AM 1 tablet was obtained the content of 35.37% and 32.62% for TG by ICP-OES, at 6 AM a percentage of 17.77% and 16.82% and for AM 7 results obtained were 8.93% for both techniques, showing that the thermogravimetry can be used to determine the percentage of calcium in tablets. The technique offers speed, economy in the use of samples and procedures eliminating the use of acid reagents in the process of the sample and efficiency results.

Conciliação entre modelos de mecanismos avançados de oxidação eletroquímica

Many pollutants dumped in waterways, such as dyes and pesticides, have become so ubiquitous that they represent a serious threat to human health. The electrochemical oxidation is presented as an alternative clean, efficient and economic degradation of wastewater containing organic compounds and a number of advantages of this technique is to just not make use of chemical reagents, since only electrical energy is consumed during the removal of pollutants organic. However, despite being a promising alternative, still needs some tweaking in order to obtain better efficiency in the elimination of persistent pollutants. Thus, this study sought a relationship between a recently discovered phenomenon that reflects the participation of dissolved oxygen in solution in the electrochemical oxidation process, as an anomaly, present a kinetic model that shows instantaneous current efficiency (ICE) above 100% limited by theory, manifested for some experiments with phenolic compounds with H2SO4 or HClO4 as supporting electrolyte with electrodes under anodic oxidation on boron doped diamond (BDD). Therefore it was necessary to reproduce the data ICE exposes the fault model, and thus the 2-naphthol was used as phenolic compound to be oxidised at concentrations of 9, 12 and 15 mmol L-1, and H2SO4 and HClO4 to 1 mol L-1 as a supporting electrolyte under a current density of 30 mA cm-2 in an electrochemical reactor for continuous flow disk configuration, and equipped with anodes DDB at room temperature (25 oC). Experiments were performed using N2 like as purge gas for eliminate oxygen dissolved in solution so that its influence in the system was studied. After exposure of the anomaly of the ICE model and investigation of its relationship with dissolved O2, the data could be treated, making it possible for confirmation. But not only that, the data obtained from eletranálise and spectroscopic analysis suggest the involvement of other strongly oxidizing species (O3 (ozone) and O radicals and O2 -), since the dissolved O2 can be consumed during the formation of new strong oxidizing species, not considered until now, something that needs to be investigated by more accurate methods that we may know a little more of this system. Currently the performance of the electrocatalytic process is established by a complex interaction between different parameters that can be optimized, so it is necessary to the implementation of theoretical models, which are the conceptual lens with which researchers see

Aplicação da técnica eletroanalítica de pulso diferencial usando o eletrodo de carbono vítreo modificado com cisteína para monitorar a degradação eletroquímica de ácido oxálico

In this paper, the technique of differential pulse voltammetry (DPV) has been studied for monitoring the concentration of oxalic acid (OA) during their electrochemical oxidation (EO) in acidic medium using platinum anode supported on titanium (Ti / Pt). The DPV was standardized and optimized using a glassy carbon electrode modified with cysteine. The modification with cysteine was developed electrochemically, forming a polymeric film on the surface of the glassy carbon electrode. The formation of the polymer film was confirmed by analysis of scanning electron microscope and atomic force microscope, confirming the modification of the electrode. The electrochemical degradation was developed using different current densities 10, 20 30 and 40 mA cm -2 electrode with Ti / Pt observing the degradation of oxalic acid, and monitored using the method of KMnO4 titration. However, the analyzes with DPV showed the same behavior elimination of oxalic acid titration. Compared with the titration method classical observed and DPV could be a good fit, confidence limits of detection and confirming the applicability of the technique electroanalytical for monitoring the degradation of oxalic acid

Caracterização e aplicação de vermicultura natural e quimicamente modificada na adsorção de azul de metileno

Chemical modification of clays has been extremely studied in the search for improvements of their properties for use in various areas, such as in combating pollution by industrial effluents and dyes. In this work, the vermiculite was chemically modified in two ways, characterized and evaluated the adsorption of methylene blue dye. First was changed with the addition of a surfactant (hexadecyltrimethylammonium bromide, BHTA) making it an organophilic clay and then by adding an acid (HCl) by acid activation. Some analyzes were performed as X-ray fluorescence (FRX), X-ray diffraction (DRX), adsorption isotherms of methylene blue dye, infrared (FTIR) , scanning electron microscopy (SEM), thermal gravimetric analysis and spectroscopy energy dispersive (EDS). Analysis by FRX of natural vermiculite indicates that addition of silicon and aluminum, clay presents in its structure the magnesium, calcium and potassium with 16 % organic matter cations. The DRX analyzes indicated that the organic vermiculite was an insertion of the surfactant in the space between the lamellae, vermiculite and acid partial destruction of the structure with loss of crystallinity. The adsorption isotherms of methylene blue showed that there was a significant improvement in the removal of dye to the vermiculite with the addition of cationic surfactant hexadecyltrimethylammonium bromide and treatment with acid using HCl 2 mol/L. In acid vermiculites subsequently treated with surfactant, the adsorption capacity increased with respect to natural vermiculite, however was much lower compared vermiculite modified with acid and surfactant separately. Only the acidic vermiculite treated with surfactant adjusted to the Langmuir model. As in the infrared spectrometry proved the characteristics of natural vermiculite. In the organic vermiculite was observed the appearance of characteristic bands of CH3, CH2, and (CH3)4N. Already on acid vermiculite, it was realized a partial destruction with decreasing intensity of the characteristic band of vermiculite that is between 1074 and 952 cm-1. In the SEM analysis, it was observed that there was partial destruction to the acid treatment and a cluster is noted between the blades caused by the presence of the surfactant. The TG shows that the higher mass loss occurs at the beginning of the heating caused by the elimination of water absorbed on the surface between layers. In the organic vermiculite also observed a loss of mass between 150 and 300 °C caused decomposition of the alkylammonium molecules (surfactants)

Melhorando a estimação de pose com o RANSAC preemptivo generalizado e múltiplos geradores de hipóteses

The camera motion estimation represents one of the fundamental problems in Computer Vision and it may be solved by several methods. Preemptive RANSAC is one of them, which in spite of its robustness and speed possesses a lack of flexibility related to the requirements of applications and hardware platforms using it. In this work, we propose an improvement to the structure of Preemptive RANSAC in order to overcome such limitations and make it feasible to execute on devices with heterogeneous resources (specially low budget systems) under tighter time and accuracy constraints. We derived a function called BRUMA from Preemptive RANSAC, which is able to generalize several preemption schemes, allowing previously fixed parameters (block size and elimination factor) to be changed according the applications constraints. We also propose the Generalized Preemptive RANSAC method, which allows to determine the maximum number of hipotheses an algorithm may generate. The experiments performed show the superiority of our method in the expected scenarios. Moreover, additional experiments show that the multimethod hypotheses generation achieved more robust results related to the variability in the set of evaluated motion directions

Revisão por pares na Revista Brasileira de Educação Especial

O artigo discute algumas questões relativas ao procedimento de revisão por pares na avaliação de originais submetidos a um periódico. Essa avaliação é aceita como necessária para assegurar a qualidade científica de artigos publicados, já que a integridade da literatura especializada é considerada essencial para o desenvolvimento da ciência. Entretanto, há críticas à revisão por pares, incluindo o longo tempo decorrido entre a entrega dos originais e a publicação, o tempo e a energia despendidos pelos autores e revisores, a subjetividade na avaliação e outros problemas que podem introduzir vieses na área. A Revista Brasileira de Educação Especial, que adota a avaliação cega por pares, tem-se mostrado relativamente versátil. O tempo médio entre o recebimento dos originais e o aceite final, depois de passar pela revisão por pares e conseqüente ajuste por parte dos autores, é de cerca de cinco meses e meio. Na literatura especializada, encontram-se sugestões como a eliminação de avaliação e publicação de todos os textos submetidos, em meios eletrônicos, menos onerosos e mais versáteis, deixando que os próprios leitores façam a sua avaliação. A análise procedida conclui que a revisão por pares é necessária. Aponta a necessidade de se realizarem discussões sobre a própria avaliação, para que se desenvolvam procedimentos mais eficientes de controle de qualidade das publicações. Sugere que a avaliação efetiva é feita pela própria comunidade acadêmica, após a publicação, mediante reconhecimento coletivo do valor científico da publicação, na forma de impacto que pode causar sobre novas investigações e construção do conhecimento na área.