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

Detecção de bactérias periodontopatogênicas cultiváveis e não cultiváveis em placas ateromatosas

Periodontal infections consist of a group of inflammatory conditions caused by microorganisms that colonize the tooth surface through the formation of dental biofilm. Chronic infections such as periodontitis have been associated to the development and progression of atherosclerosis. AIM: Detect cultivatable and non-cultivatable periodontopathogenic bacteria in atheromatous plaques; search for factors associated to the presence of these bacteria in the atheromatous plaques and characterize the presence of cultivatable and non-cultivatable bacteria in these plaques. METHODOLOGY: A cross-sectional study was performed with a sample of 30 patients diagnosed with atherosclerosis in the carotid, coronary or femoral arteries and surgically treated with angioplasty and stent implant, bypass or endarterectomy. The plaques were collected during surgery and analyzed using the PCR molecular technique for the presence of the DNA of the cultivatable bacteria Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Treponema denticola and of the non-cultivatable Synergistes phylotypes. The patients were examined in the infirmary, after the surgery, where they also responded to a questionnaire aimed at determining factors associated to the presence of periodontopathogenic bacteria in the atheromatous plaques. RESULTS: All patients with tooth (66,7%) possessed disease periodontal, being 95% severe and 65% widespread. No periodontopathogenic bacteria were found in the atheromatous plaques. However, four samples (13.3%) were positive for the presence of bacteria. Of these, three participants were dentate, being two carriers of widespread severe chronic periodontite and one of located severe chronic periodontitis. None of them told the accomplishment of procedures associated to possible bacteremia episodes, as treatment endodontic, extraction the last six months or some procedure surgical dental. CONCLUSION: The periodontopathogenic bacteria studied were not found in the atheromatous plaques, making it impossible to establish the prevalence of these pathogens or the factors associated to their presence in plaques, the detection of positive samples for bacteria suggests that other periodontal and non-periodontal pathogens be studied in an attempt at discovering the association or not between periodontal disease and/or others infections and atherosclerosis, from the presence of these bacteria in atheromas

Estudo da detecção do DNA do papiloma vírus humano (HPV) e da expressão imuno-histoquímica de proteína do ciclo celular no carcinoma epidermóide oral

Oral squamous cell carcinoma (OSCC) is the most common malignancy in oral cavity and human papillomavirus (HPV) may have an important role in its development. The aim of this experiment was to investigate the HPV DNA and viral types in 90 cases of OSCC. Moreover, a comparative analysis between the cases of OSSC with and without HPV DNA was performed by using cell cycle markers p21 and pRb in order to detect a possible correlation of these proteins and HPV infection. DNA was extracted from paraffin embedded tissue and amplified by PCR (polymerase chain reaction) with primers PCO3+ e PCO4+ for a fragment of human β-globin gene. After this procedure, PCR for HPV DNA detection was realized using a pair of generic primers GP5+ e GP6+. Immunohistochemical study was performed by streptoavidin-biotin technique and antibodies against p21 and pRb proteins were employed. Eighty-eight cases were positive for human β-globin gene and HPV DNA was found in 26 (29.5%) of then. It could not be detected significant correlation between HPV and age, sex and anatomical sites of the lesion. The most prevalent viral type was HPV 18 (80.8%). Regarding the immunohistochemical analysis, it was detected significant association between HPV presence and pRb immunoexpression (p=0,044), nevertheless, the same was not observed in relation to p21 protein (p =0,416). It can be concluded that the low detection of HPV DNA in OSCC by the present experiment suggests a possible role of the virus in the development and progression in just a subset of this disease

Detecção de cor e forma por Callithrix jacchus em cativeiro

Callithrix jacchus studies involving differences between the sexes regarding the performance on tasks food still offers room for the investigation of some factors, among them there is the differences in color vision, which can directly influence the detection of visual clues on food items. This study aimed to analyze the performance of C. jacchus in tasks involving detection of food items. Some factors were analyzed such as the differences in performance between the sexes and behavioral categories present during the task. There were no differences in performance between the animals in carrying out the task, for all situations presented, examining the behavioral categories observed. The fact of the task to be very simple might have influenced the results, and it was not possible to observe differences in performance. Males and females showed the same performance in all analyzed situations. The sex differences were not found possibly due to the influence of external factors, such as the structure of the experimental apparatus. The animals are more efficient in carrying out the task during the morning, in comparison to the afternoon. The light may have been one of the factors that influenced these results. Due to the influence of other factors that probably contributed to these results, we believe that different results can be found in future work

Detecção de predadores por dicromatas e tricromatas humanos e a sua implicação na evolução da visão de cores em primatas

Among placental mammals, primates are the only ones to present trichromatic color vision. However, the distribution of trichromacy among primates is not homogeneous: Old World primates shows an uniform trichromacy (with all individuals being trichromats) and New World primates exhibit a color vision polymorphism (with dichromatic males and dichromatic or trichromatic females). Visual ecology studies have investigated which selective pressures may have been responsible for the evolution of trichromacy in primates, diverging from the dichromat standard found in other mammals. Cues associated with foraging and the socio-reproductive status were analyzed, indicating a trichromatic advantage for the rapid detection of visually conspicuous objects against a green background. However, dichromats are characterized by an efficient capture of cryptic and camouflaged stimuli. These advantages regarding phenotype may be responsible for the maintenance of the visual polymorphism in New World primates and for the high incidence of color blindness in humans (standing around 8% in Caucasian men). An important factor that has not yet been experimentally taken into account is the predation risk and its effect on the evolution of trichromacy in primates. To answer this question, we prepared and edited pictures of animals with different coats: oncillas (Leopardus spp.), puma (Puma concolor) and ferret (Galictis cuja). The specimens were taxidermized and the photographs were taken in three different vegetation scenarios (dense forest, cerrado and grassland). The images of the predators were manipulated so that they fit into two categories of stimulus size (small or large). After color calibration and photo editing, these were presented to 40 humans (20 dichromats and 20 trichromats) by a computer program, which presented a set of four photos at a time (one picture containing the taxidermized animal amid the background vegetation and three depicting only the background vegetation) and recorded the response latency and success rate of the subjects. The results show a trichromatic advantage in detecting potential predators. The predator detection was influenced by the background, the predator species, the dimension of the stimulus and the observer s visual phenotype. As humans have a high rate of dyschromatopsias, when compared to wild Catarrhini or human tribal populations, it is possible that the increased rate of dichromats is a result of reduced pressure for rapid predator detection. Since our species came to live in more cohesive groups and resistant to attack by predators, with the advent of agriculture and the formation of villages, it is possible that the lower risk of predation has reduced the selection in favor of trichromats

Detecção de metamidofós em solos por métodos ecotoxicológico e cromatografia líquida acoplada à espectrometria de massas sequencial (LC-MS/MS)

Soil contamination by pesticides is an environmental problem that needs to be monitored and avoided. However, the lack of fast, accurate and low cost analytical methods for discovering residual pesticide in complex matrices, such as soil, is a problem still unresolved. This problem needs to be solved before we are able to assess the quality of environmental samples. The intensive use of pesticides has increased since the 60s, because the dependence of their use, causing biological imbalances and promoting resistance and recurrence of high populations of pests and pathogens (upwelling). This has contributed to the appearance of new pests that were previously under natural control. To develop analytical methods that are able to quantify residues pesticide in complex environment. It is still a challenge for many laboratories. The integration of two analytical methods one ecotoxicological and another chemical demonstrates the potential for environmental analysis of methamidophos. The aim of this study was to evaluate an ecotoxicological method as "screening" analytical methamidophos in the soil and perform analytical confirmation in the samples of the concentration of the analyte by chemical method LC-MS/MS In this work we tested two soils: a clayey and sandy, both in contact with the kinetic methamidophos model followed pseudo-second order. The clay soil showed higher absorption of methamidophos and followed the Freundlich model, while the sandy, the Langmuir model. The chemical method was validated LC-MS/MS satisfactory, showing all parameters of linearity, range, precision, accuracy, and sensitivity adequate. In chronic ecotoxicological tests with C. dubia, the NOEC was 4.93 and 3.24 for ng L-1 of methamidophos to elutriate assays of sandy and clay soils, respectively. The method for ecotoxicological levels was more sensitive than LC-MS/MS detection of methamidophos, loamy and sandy soils. However, decreasing the concentration of the standard for analytical methamidophos and adjusting for the validation conditions chemical acquires a limit of quantification (LOQ) in ng L-1, consistent with the provisions of ecotoxicological test. The methods described should be used as an analytical tool for methamidophos in soil, and the ecotoxicological analysis can be used as a "screening" and LC-MS/MS as confirmatory analysis of the analyte molecule, confirming the objectives of this work

Detecção e recuperação de falhas para a máquina de redução de grafos PEWS-AM

Web services are software units that allow access to one or more resources, supporting the deployment of business processes in the Web. They use well-defined interfaces, using web standard protocols, making possible the communication between entities implemented on different platforms. Due to these features, Web services can be integrated as services compositions to form more robust loose coupling applications. Web services are subject to failures, unwanted situations that may compromise the business process partially or completely. Failures can occur both in the design of compositions as in the execution of compositions. As a result, it is essential to create mechanisms to make the implementation of service compositions more robust and to treat failures. Specifically, we propose the support for fault recovery in service compositions described in PEWS language and executed on PEWS-AM, an graph reduction machine. To support recovery failure on PEWS-AM, we extend the PEWS language specification and adapted the rules of translation and reduction of graphs for this machine. These contributions were made both in the model of abstract machine as at the implementation level

Sistema de aquisição de dados para estudos de transiente de pressão e detecção de vazamentos em oleodutos

In February 2011, the National Agency of Petroleum, Natural Gas and Biofuels (ANP) has published a new Technical Rules for Handling Land Pipeline Petroleum and Natural Gas Derivatives (RTDT). Among other things, the RTDT made compulsory the use of monitoring systems and leak detection in all onshore pipelines in the country. This document provides a study on the method for detection of transient pressure. The study was conducted on a industrial duct 16" diameter and 9.8 km long. The pipeline is fully pressurized and carries a multiphase mixture of crude oil, water and natural gas. For the study, was built an infrastructure for data acquisition and validation of detection algorithms. The system was designed with SCADA architecture. Piezoresistive sensors were installed at the ends of the duct and Digital Signal Processors (DSPs) were used for sampling, storage and processing of data. The study was based on simulations of leaks through valves and search for patterns that characterize the occurrence of such phenomena

Detecção e diagnóstico de falhas não-supervisionados baseados em estimativa de densidade recursiva e classificador fuzzy auto-evolutivo

In this work, we propose a two-stage algorithm for real-time fault detection and identification of industrial plants. Our proposal is based on the analysis of selected features using recursive density estimation and a new evolving classifier algorithm. More specifically, the proposed approach for the detection stage is based on the concept of density in the data space, which is not the same as probability density function, but is a very useful measure for abnormality/outliers detection. This density can be expressed by a Cauchy function and can be calculated recursively, which makes it memory and computational power efficient and, therefore, suitable for on-line applications. The identification/diagnosis stage is based on a self-developing (evolving) fuzzy rule-based classifier system proposed in this work, called AutoClass. An important property of AutoClass is that it can start learning from scratch". Not only do the fuzzy rules not need to be prespecified, but neither do the number of classes for AutoClass (the number may grow, with new class labels being added by the on-line learning process), in a fully unsupervised manner. In the event that an initial rule base exists, AutoClass can evolve/develop it further based on the newly arrived faulty state data. In order to validate our proposal, we present experimental results from a level control didactic process, where control and error signals are used as features for the fault detection and identification systems, but the approach is generic and the number of features can be significant due to the computationally lean methodology, since covariance or more complex calculations, as well as storage of old data, are not required. The obtained results are significantly better than the traditional approaches used for comparison

Aplicação do Sensoriamento Remoto e do Sistema de Informações Geográficas na Detecção de Manchas de Óleo na Região do Pólo de Exploração de Guamaré, R.N.

Objective to establish a methodology for the oil spill monitoring on the sea surface, located at the Submerged Exploration Area of the Polo Region of Guamaré, in the State of Rio Grande do Norte, using orbital images of Synthetic Aperture Radar (SAR integrated with meteoceanographycs products. This methodology was applied in the following stages: (1) the creation of a base map of the Exploration Area; (2) the processing of NOAA/AVHRR and ERS-2 images for generation of meteoceanographycs products; (3) the processing of RADARSAT-1 images for monitoring of oil spills; (4) the integration of RADARSAT-1 images with NOAA/AVHRR and ERS-2 image products; and (5) the structuring of a data base. The Integration of RADARSAT-1 image of the Potiguar Basin of day 21.05.99 with the base map of the Exploration Area of the Polo Region of Guamaré for the identification of the probable sources of the oil spots, was used successfully in the detention of the probable spot of oil detected next to the exit to the submarine emissary in the Exploration Area of the Polo Region of Guamaré. To support the integration of RADARSAT-1 images with NOAA/AVHRR and ERS-2 image products, a methodology was developed for the classification of oil spills identified by RADARSAT-1 images. For this, the following algorithms of classification not supervised were tested: K-means, Fuzzy k-means and Isodata. These algorithms are part of the PCI Geomatics software, which was used for the filtering of RADARSAT-1 images. For validation of the results, the oil spills submitted to the unsupervised classification were compared to the results of the Semivariogram Textural Classifier (STC). The mentioned classifier was developed especially for oil spill classification purposes and requires PCI software for the whole processing of RADARSAT-1 images. After all, the results of the classifications were analyzed through Visual Analysis; Calculation of Proportionality of Largeness and Analysis Statistics. Amongst the three algorithms of classifications tested, it was noted that there were no significant alterations in relation to the spills classified with the STC, in all of the analyses taken into consideration. Therefore, considering all the procedures, it has been shown that the described methodology can be successfully applied using the unsupervised classifiers tested, resulting in a decrease of time in the identification and classification processing of oil spills, if compared with the utilization of the STC classifier

Detecção de áreas degradadas na sub-bacia hidrográfica do rio taperoá/pb, utilizando parâmetros físicos dos sensores modis/terra e tm/landsat

This study includes the results of the analysis of areas susceptible to degradation by remote sensing in semi-arid region, which is a matter of concern and affects the whole population and the catalyst of this process occurs by the deforestation of the savanna and improper practices by the use of soil. The objective of this research is to use biophysical parameters of the MODIS / Terra and images TM/Landsat-5 to determine areas susceptible to degradation in semi-arid Paraiba. The study area is located in the central interior of Paraíba, in the sub-basin of the River Taperoá, with average annual rainfall below 400 mm and average annual temperature of 28 ° C. To draw up the map of vegetation were used TM/Landsat-5 images, specifically, the composition 5R4G3B colored, commonly used for mapping land use. This map was produced by unsupervised classification by maximum likelihood. The legend corresponds to the following targets: savanna vegetation sparse and dense, riparian vegetation and exposed soil. The biophysical parameters used in the MODIS were emissivity, albedo and vegetation index for NDVI (NDVI). The GIS computer programs used were Modis Reprojections Tools and System Information Processing Georeferenced (SPRING), which was set up and worked the bank of information from sensors MODIS and TM and ArcGIS software for making maps more customizable. Initially, we evaluated the behavior of the vegetation emissivity by adapting equation Bastiaanssen on NDVI for spatialize emissivity and observe changes during the year 2006. The albedo was used to view your percentage of increase in the periods December 2003 and 2004. The image sensor of Landsat TM were used for the month of December 2005, according to the availability of images and in periods of low emissivity. For these applications were made in language programs for GIS Algebraic Space (LEGAL), which is a routine programming SPRING, which allows you to perform various types of algebras of spatial data and maps. For the detection of areas susceptible to environmental degradation took into account the behavior of the emissivity of the savanna that showed seasonal coinciding with the rainy season, reaching a maximum emissivity in the months April to July and in the remaining months of a low emissivity . With the images of the albedo of December 2003 and 2004, it was verified the percentage increase, which allowed the generation of two distinct classes: areas with increased variation percentage of 1 to 11.6% and the percentage change in areas with less than 1 % albedo. It was then possible to generate the map of susceptibility to environmental degradation, with the intersection of the class of exposed soil with varying percentage of the albedo, resulting in classes susceptibility to environmental degradation

Método híbrido para detecção e diagnóstico de falhas baseado em resíduos

The detection and diagnosis of faults, ie., find out how , where and why failures occur is an important area of study since man came to be replaced by machines. However, no technique studied to date can solve definitively the problem. Differences in dynamic systems, whether linear, nonlinear, variant or invariant in time, with physical or analytical redundancy, hamper research in order to obtain a unique solution . In this paper, a technique for fault detection and diagnosis (FDD) will be presented in dynamic systems using state observers in conjunction with other tools in order to create a hybrid FDD. A modified state observer is used to create a residue that allows also the detection and diagnosis of faults. A bank of faults signatures will be created using statistical tools and finally an approach using mean squared error ( MSE ) will assist in the study of the behavior of fault diagnosis even in the presence of noise . This methodology is then applied to an educational plant with coupled tanks and other with industrial instrumentation to validate the system.

Proteção diferencial de transformadores de potência utilizando a transformada Wavelet

About 10% of faults involving the electrical system occurs in power transformers. Therefore, the protection applied to the power transformers is essential to ensure the continuous operation of this device and the efficiency of the electrical system. Among the protection functions applied to power transformers, the differential protection appears as one of the main schemes, presenting reliable discrimination between internal faults and external faults or inrush currents. However, when using the low frequency components of the differential currents flowing through the transformer, the main difficulty of the conventional methods of differential protection is the delay for detection of the events. However, internal faults, external faults and other disturbances related to the transformer operation present transient and can be appropriately detected by the wavelet transform. In this paper is proposed the development of a wavelet-based differential protection for detection and identification of external faults to the transformer, internal faults, and transformer energizing by using the wavelet coefficient energy of the differential currents. The obtained results reveal the advantages of using of the wavelet transform in the differential protection compared to conventional protection, since it provides reliability and speed in detection of these events.

Estudo sobre a detecção de invariância de escala discreta em sistemas com criticalidade auto-organizada

Recent studies have shown evidence of log-periodic behavior in non-hierarchical systems. An interesting fact is the emergence of such properties on rupture and breakdown of complex materials and financial failures. These may be examples of systems with self-organized criticality (SOC). In this work we study the detection of discrete scale invariance or log-periodicity. Theoretically showing the effectiveness of methods based on the Fourier Transform of the log-periodicity detection not only with prior knowledge of the critical point before this point as well. Specifically, we studied the Brazilian financial market with the objective of detecting discrete scale invariance in Bovespa (Bolsa de Valores de S˜ao Paulo) index. Some historical series were selected periods in 1999, 2001 and 2008. We report evidence for the detection of possible log-periodicity before breakage, shown its applicability to the study of systems with discrete scale invariance likely in the case of financial crashes, it shows an additional evidence of the possibility of forecasting breakage

Elaboração de pós de ferrita de cobalto, CoFe2O4, nanoestruturados e hierarquizados: influência da morfologia sobre a detecção e oxidação catalítica de gases poluentes

This thesis is part of research on new materials for catalysis and gas sensors more active, sensitive, selective. The aim of this thesis was to develop and characterize cobalt ferrite in different morphologies, in order to study their influence on the electrical response and the catalytic activity, and to hierarchize these grains for greater diffusivity of gas in the material. The powders were produced via hydrothermal and solvothermal, and were characterized by thermogravimetric analysis, X-ray diffraction, scanning electron microscopy, transmission electron microscopy (electron diffraction, highresolution simulations), and energy dispersive spectroscopy. The catalytic and electrical properties were tested in the presence of CO and NO2 gases, the latter in different concentrations (1-100 ppm) and at different temperatures (room temperature to 350 ° C). Nanooctahedra with an average size of 20 nm were obtained by hydrothermal route. It has been determined that the shape of the grains is mainly linked to the nature of the precipitating agent and the presence of OH ions in the reaction medium. By solvothermal method CoFe2O4 spherical powders were prepared with grain size of 8 and 20 nm. CoFe2O4 powders exhibit a strong response to small amounts of NO2 (10 ppm to 200 ° C). The nanooctahedra have greater sensitivity than the spherical grains of the same size, and have smaller response time and shorter recovery times. These results were confirmed by modeling the kinetics of response and recovery of the sensor. Initial tests of catalytic activity in the oxidation of CO between temperatures of 100 °C and 350 °C show that the size effect is predominant in relation the effect of the form with respect to the conversion of the reaction. The morphology of the grains influence the rate of reaction. A higher reaction rate is obtained in the presence of nanooctahedra. In order to improve the detection and catalytic properties of the material, we have developed a methodology for hierarchizing grains which involves the use of carbonbased templates.