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

Cartografia temporal e análise geoambiental da dinâmica da foz do Rio Piranhas-Açu, regiao de Macau-RN, com base em imagens Landast 5-MT

This work embraces the application of Landsat 5-TM digital images, comprising August 2 1989 and September 22 1998, for temporal mapping and geoenvironmental analysis of the dynamic of Piranhas-Açu river mouth, situated in the Macau (RN) region. After treatment using several digital processing techniques (e.g. colour composition in RGB, ratio of bands, principal component analysis, index methods, among others), it was possible to generate several image products and multitemporal maps of the coastal morphodynamics of the studied area. Using the image products it was possible the identification and characterization of the principal elements of interest (vegetation, soil, geology and water) in the surface of the studied area, associating the spectral characteristics of these elements to that presented by the image products resulting of the digital processing. Thus, it was possible to define different types of soils: Amd, AQd6, SK1 and LVe4; vegetation grouping: open arboreal-shrubby caatinga, closed arborealshrubby caatinga, closed arboreal caatinga, mangrove vegetation, dune vegetation and areas predominately constituted by juremas; geological units: quaternary units beach sediments, sand banks, dune flats, barrier island, mobile dunes, fixed dunes, alluvium, tidal and inundation flats, and sandy facies of the Potengi Formation; tertiary-quaternary units Barreiras Formation grouped to the clayey facies of the Potengi Formation, Macau Formation grouped to the sediments of the Tibau Formation; Cretaceous units Jandaíra Formation; moreover it was to identify the sea/land limit, shallow submersed areas and suspended sediments. The multitemporal maps of the coastal morphodynamics allowed the identification and a semi-quantitative evoluation of regions which were submitted to erosive and constructive processes in the last decade. This semi-quantitative evoluation in association with an geoenvironmental characterization of the studied area are important data to the elaboration of actions that may minimize the possible/probable impacts caused by the implantation of the Polo Gas/Sal and to the monitoring of areas explorated by the petroleum and salt industries

Aids em idosos no Brasil no período de 2000 a 2012: uma análise de série temporal e dos fatores contextuais associados

The characteristics profile of individuals who develop AIDS in Brazil has changed over time. Among these modifications, a worrying finding is the increased incidence of AIDS in the elderly across the country. But, however, is not yet clear whether the increase in AIDS cases is sufficient to produce a change in the trend of measures in recent years in the Brazilian states, and this increase has an effect from the socioeconomic and demographic indicators. In this sense, the objective of this study is to analyze the AIDS incidence rates among the elderly in Brazil and its effect on socioeconomic and demographic inequalities in the period 2000 to 2012. This is an ecological time-series study to meet behavior of the time series of the incidence rates of AIDS in the elderly from 2000 to 2012. the rates were calculated using the secondary data from Diseases Information System Notification and the Brazilian Institute of Geography and Statistics. Data were analyzed statistically to know the trends in incidence rates, by polynomial regression model and joinpoint log-linear regression model, but also the simple linear regression analysis to find the relationship of trends with variables socioeconomic and demographic. SPSS 20.0® and Joinpoint 4.1.1 programs were used. All tests were carried out considering a significance of 5%. After the analysis, in Brazil were reported 62,052 new cases of AIDS in the elderly from 2000 to 2012. During this period, a significant increase was found for males, both aged 50-59 years (APPC: 3.46 %, p <0.001), such as above 59 years (AAPC: 4.38%; p <0.001). For females, the increase was significant and has the largest increments in the time series, when compared to males in both age groups (AAPC: 4.62%, p <0.001 and AAPC: 6.53%; p <0.001) respectively. The largest increases are observed in women and in the states of North and Northeast. In the Southeast Region is observed stabilization of rates throughout the series. The reason of trends between the sexes had a significant reduction, but also an approach in both age groups of the study, reaching a ratio of 1.7 males for every female in the youngest age group. The trends were related to illiteracy rates, with increasing social inequality and the lowest human development in the Brazilian states. We conclude that in Brazil the incidence of AIDS in the elderly follows an increasing trend in individuals over 50 years. Noteworthy are the highest rates of study in women and in the states of North and Northeast. In this sense, the country needs to enhance policies towards older people with STD / AIDS, training health professionals and developing effective measures for the prevention and early diagnosis of infected people, especially in places with limited resources and high social inequality. In the long term, it is developing new studies to understand whether the measures taken were effective in reducing the trends identified in this study.