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

Expressão imuno-histoquímica da calponina em glândula parótida de rato após obstrução do ducto excretor principal

Glandular atrophy is one of several alterations which can aflict the salivary glands, caused generally by obstructive lesions such as sialo1ithiasis, infections or compression by neoplastic processes arnong others. In this work, a morphological and immunohistochemical study was carried out in rat parotid glands, which were submitted to obstruction of the main excretory duct suffering atrophy at varied time intervals, with the aim of appraising the behavior of myoepithelial cells during the process of glandular atrophy. It was analized the immunohistochemical expression of calponin which detects myoepithelial cells in the parotids of 28 animaIs, which were divided into 7 groups, each one made up of 4 rats, afier the ductal ligature procedure, in the following time intervals: zero hour (control), 24 hours, 7, 15, 21, 30 and 60 days. Analysis of the immunohistochemicaI profile was carried through in which the calponin expression was veritied through its distribution pattem and numericaI index. All specimens exhibited positivity for calponin in myoepithelial cells which were distributed around the acini and the ductaI structures, a small number of positiveIy marked cells being detected in the controI group and in the 24-hour group when compared to subsequent ones, where it was perceived a Iarge increase in the number of positiveIy marked cens, mainly surrounding the ductiform structures which originated during the obstruction time. Upon application of statistical tests it was verified that the rise in the number the myoepithelial positive cells for calponin, when the control groups (zero hour) was compared to the 7, 15,21, 30 and 60-day groups afier obstruction, was statistica1ly significant. It was concluded then that the detected rise probably carne about due to an elevation in the rate of proliferation of the myoepitheliaI cells subsequent to the ductal obstruction, associated with a growing resistance of these cells to glandular atrophy.

Saguis (Callithrix jacchus) sob ciclo claro-escuro de 21 h: um modelo de dessincronização forçada em primata diurno

The circadian system consists of multiple oscillators organized hierarchically, with the suprachiasmatic nucleus (SCN) as the master oscillator to mammalians. There are lots of evidences that each SCN cell is an oscillator and that entrainment depends upon coupling degree between them. Knowledge of the mechanism of coupling between the SCN cells is essential for understanding entrainment and expression of circadian rhythms, and thus promote the development of new treatments for circadian rhythmicity disorders, which may cause various diseases. Some authors suggest that the dissociation model of circadian rhythm activity of rats under T22, period near the limit of synchronization, is a good model to induce internal desynchronization, and in this way, enhance knowledge about the coupling mechanism. So, in order to evaluate the pattern of the motor activity circadian rhythm of marmosets, Callithrix jacchus, in light-dark cycles at the lower limit of entrainment, two experiments were conducted: 1) 6 adult females were submitted to the LD symmetric cycles T21, T22 and T21.5 for 60, 35 and 48 days, respectively; 2) 4 male and 4 female adults were subjected to T21 for 24 days followed by 18 days of LL, and then back to T21 for 24 days followed by 14 days of LL. Vocalizations of all animals and motor activity of each one of them were continuously recorded throughout the experiments, but the vocalizations were recorded only in Experiment 1. Under the Ts shorter than 24 h, two simultaneous circadian components appeared in motor activity, one with the same period of LD cycle, named light-entrained component, and the other in free-running, named non-light-entrained component. Both components were displayed for all the animals in T21, five animals (83.3%) in T21.5 and two animals (33.3%) in T22. For vocalizations both components were observed under the three Ts. Due to the different characteristics of these components we suggest that dissociation is result of partial synchronization to the LD cycle, wherein at least one group oscillator is synchronized to the LD by relative coordination and masking processes, while at least another group of oscillators is in free-running, but also under the influence of masking by the LD. As the T21 h was the only cycle able to promote the emergence of both circadian components in circadian rhythms of all Callithrix jacchus, this was then considered the lower entrainment limit of LD cycle promoter of dissociation in circadian rhythmicity of this species, and then suggested as a non-human primate model for forced desynchronization

Caracterização da neofobia alimentar em crianças de três a seis anos

Alimentation is essential in life. Concerning omnivores, characterized by the necessity of a varied diet to satisfy their metabolic needs, it is extremely advantageous the assumption of new foods. However, the assumption of new unknown foods is, potentially dangerous, because of possible intoxications. In this sense, one of the most important behaviors related to reducing risks is the so called food neophobia, characterized by the rejection of new foods and/or an ingestion of very little amounts. The aim of the present study was to investigate if age, sex and socio-economical status were able to influence food neophobia. The neophobia has been described in a range of 3-6 years old children taken both from public and private schools within the city of Natal-RN. Four different type of ice-creams, each one characterized by a different flavor, have been utilized. Two flavors were known to the young and the remaining two flavor were new. We didn't find significant differences between the investigated variables. However, the exploitation of data from the survey conducted showed that the ease or not to accept new foods obtained, was correlated with the variables under the same guidelines observed in literature. Aspects related to the stimulus used probably eased the neophobic answer. Then, it is suggested that the food neophobia can be influenced by sex, age and socioeconomic factors of individuaIs. Neophobia tends to be more common in girls, with ages between three to four years old and with a low leveI socioeconomic. In this sense, given the importance of kid neophobic reaction to the development of dietary patterns of other life's stages, it is necessary to make further studies to better explain this phenomenon. Given the pivotal role of food neophobia to the development of alimentary habits within all ages of life, other studies will be necessary for a better comprehension of such phenomena. Key-words: food neophobia; Evolutionary Psychology;children food intake; diet restriction; children's diet development

Caracterização citoarquitetônica, neuroquímica e de aferência óptica do complexo parabraquial do sagui (Callithrix jacchus)

The parabrachial complex (PB) is an area of the brainstem responsible for the processing and transmission of essential physiologic information for the survival of the organisms. This region is subdivided in approximately nine subregions, considering morphology, cytoarchitectural and functional characteristic. Its neurons have an extensive network of connections with other regions of the nervous system. The objective in this work was to map the retinal projection to the PB and make a citoarchitectonic and neurochemical characterization of this region in the common marmoset (Callithrix jacchus), a primate of the New World. The retinal projections were mapped by anterograde transport of the choleric toxin subunit b (CTb). The citoarchitecture was described through the Nissl method, and the neurochemical characterization was made through immunohistochemical technique to the some neurotransmitters and neuroactives substances present in this neural center. In marmoset PB, in the coronal sections labeled by Nissl method, we found a similar pattern to that evidenced in other animal species. The immunoreactivity against CTb was verified in the PBMv in fibers/terminal, characterizing such as retinal innervations in this area. The immunohistochemical technique reveled that the PB contain cells, fibers and/or terminals immunoreactives to the neuronal nuclear protein, Choline acetyl transferase, nitric oxide synthase, serotonin, enkephalin, substance P, Calcium-binding proteins (calbindin, calretinin e parvalbumin), and glial fibrillary acidic protein. The histochemical technique reveled cells and fibers NADPH-diaphorase reactive. Each one of those substances presented a characteristic pattern of demarcation in PB, and some serve as specific markers of subregions

Estampa temporal na memória de reconhecimento em estudantes universitários

Several studies have shown that there is a circadian modulation of explicit memory. This modulation can occur independently in each one of the mnemonic processes. The aim of this study was to evaluate the influence of time of training on short-term memory (STM) and long-term memory (LTM), using a recognition task. Moreover, a possible circadian modulation in retrieving was investigated when this process matched the acquisition hour (time stamp). The chronotype variable was also considered. Fifty-seven undergraduate students aging between 18 and 25 years (21,72 ± 2,14; 28 ♂) participated in this study. In the training phase (acquisition) the subjects heard a ten word list. Following this, they answered a recognition test to evaluate STM and one week later they answered a recognition test to evaluate LTM. In each chronotype, the subjects were divided in groups according to the training hour, part of them in the morning and the other in the afternoon. One week later some of the subjects in each group underwent LTM testing in the morning and others in the afternoon. When the subjects performances were analyzed together, independently of the chronotypes, a training hour effect was found in the LTM. The subjects trained in the afternoon had better performance. No time of day effect was found in the STM and in retrieving from the LTM. However, the morning types who were trained and tested in the same hour had a better performance in the LTM when compared to morning types trained and tested in different hours. This effect did not occur when the other chronotypes were analyzed. The circadian modulation seems to occur at least in two different ways. First, there is a circadian modulation in the acquisition/consolidation processes, with a better performance occuring in the afternoon. Secondly, there is a modulation in the retrieval mnemonic process, called time stamp phenomenon. This phenomenon, that occurred in the morning types, is showed for the first time in humans

Aspectos do cuidado cooperativo em dois grupos de Callithrix jacchus selvagens

The Callitrichidae family presents cooperative breeding, where breeders and non breeders take care of the offspring. The aspects of care analyzed in this study were infant carrying, supervision, proximity and food transfer. Three sets of infants from two wild groups of Callithrix jacchus were studied in the environments of Caatinga (Assu group) and Atlantic Forest (Jundiaí group). The methods used in the study were instantaneous focal sampling (infant carrying, supervision and proximity) and continuous focal sampling (food transfer). In the two sets observed in Assu group, the father carried and transferred food to infants more than the rest of the group. The biggest contribution in supervision was from the father and from another adult male. The members that remained in proximity to the infants in both groups were the younger in the groups (juveniles and sub-adults). In the Jundiaí group, the father and the adult male helper of the group were the main caregivers; one of the sub-adult females was responsible for supervision of the infants. With the disappearance of the reproductive male and one of the sub-adults females in 3º month of infants life, the care was redistributed and the only adult male left in the group was the animal that contributed more in provisioning of the infant. In the Assu group, there were adult females in its composition which were involved in agonistic interactions with breeders and adult males, and seemed to influence their low participation in care. Food transfer initiates early in the development of the infants, as a way to encourage nutritional independence. Different types of food transfer (active food transfer, food steal, food steal attempt, passive food transfer and food handling) were observed in the study and frequency of each one varied with developmental phase and tolerance by the members. One relevant data of the study was the presence of active food transfer in Assu group, since in literature there are very few registers of this type of transfer for this species. It is important that groups from distinct environments and composition be studied for a better understanding of the dynamics of infants development

A experiência de adolescentes portadores de diabetes Mellitus Tipo 1: seus significados e sentido

Worldwide, the diabetes mellitus is considered a serious problem of public health; it also involves high costs for its treatment and its complications. Even though the onset of diabetes mellitus type 1 is on 5% to 10% of the diabetic population, it is the most aggressive type of anifestation; furthermore, it is the commonest chronic disease in childhood and adolescence. Such data show the importance of understanding the development of this disease from the moment the individual perceives it, according to their experience, observing the complexity of the phenomenon thoroughly. This study aimed to comprehend the experience of the adolescent with onset of diabetes mellitus type 1 in order to learn how these young individuals perceive and reason their place in the world when they experience this illness. The methodological strategy was based on qualitative research of phenomenological inspiration. We applied the main ideas of heideggerian ontology and some concepts of existential phenomenological psychology to guide our reflexive path. The participants were 10 young individuals (06 female and 04 male), ages between 15 and 18, under treatment in the city of Recife, Pernambuco. The methodological resource was the narrative, which allowed us access to the adolescents experience. The comprehension of the reports was based on the heideggerian hermeneutics, whose priority is the interpretation of the meaning expressed by the participants speech. The results showed that factors such as social, family, economic and cultural context greatly influence the perception and the way the individuals deal with, perceive and reason their experience. We noticed that the diabetes type 1 may often represent a limitation, but it does not make it impossible to be part of the world with others. The data show a significant affective ambivalence regarding the content more directly related to the disease. This demonstrates the unique way which each one gives meaning and reasons their condition of existence. This research points out the need to understand the complexity of this phenomenon in a more complete manner, considering the adolescent in their timely and historical context. We believe we can encourage thinking that may become actions which are more fit to the reality researched regarding all the parties involved with the subject, such as health professionals, adolescents, family, support programs and public health policies

Compreendendo a experiência de pessoas portadoras de transtorno mental: histórias de vidas severinas repletas de possibilidades

This study aims to understand the experience of people suffering from mental disorder. The patients are enrolled in a mental health ambulatory clinic in the city of Natal (RN). Mental disorders are growing rapidly in the contemporary world and are a source of intense mental suffering. Besides patients being strongly marked by a history of isolation and prejudice, they have been the target of real atrocities committed in the name of preservation of a supposed normality. The understanding and treatment of this disorder is influenced by cultural and historical inferences, depending on the period in which it is experienced. Semi-directed Interviews were conducted with a group of users, with the emphasis on giving voice to their uniqueness and individuality, highlighting how each one perceives his or her own experience. These were recorded and later transcribed by identifying the core of meanings. The results were analyzed under the gaze of the Humanist Phenomenology Existential perspective, which aims to unravel the phenomenon, without truths from volatility, highlighting the existence of the mental disorder as a way of living, being permeated by suffering mental and influenced by social problems, assuming contours very particular to each individual. Some progress has been perceived, even by users, with respect to the change of paradigm in the way of care, but still there is a consistent emphasis on medical and drug use. The changes point to the need for offering services to replace the asylum hospital model, and in addition to accept the bearer of mental disorder as a citizen, a bearer of rights who should be accepted and respected by society. Despite the pain expressed and its close liaison with suicide, their reports are full of perspectives and attitudes of confrontation facing life, pointing to new possibilities to be, recreating itself

Por trás das grades :reflexões sobre a experiência do ser apenado na situação de privação de liberdade

This research has resulted of investigations appeared from our work experience as psychologist in a prison of the state Rio Grande do Norte. It deals with the meeting with prisoner in the search for the rescue of the human being dignity, discovering the being that finds annulled by backwards of the prison. The current debate on the prisoners in Brazil has as focus the creation of efficient strategies in the combat to the criminal acts, not being worried in understanding the historical and social conditions in the context in which such acts develop. Our objective was to reach an understanding of the experience of the prisoner in the situation of freedom deprivation in Natal (RN) city. Its significant social relevance meets in emphasized thematic which intends to be revealing of the not-said one of these citizens. The epistemological space and the vision of adopted human being support in the humanist ideas of the Centered Approach in the Person and construct self, central nucleus of the personality theory proposal by Carl Rogers. We opt to the phenomenological method as way of access to the singularity of the experience of each one of the participants, supporting us in the strategy of the narrative as expression of the lived world. The results had pointed that the investigated experience is through suffering, that is, the way to be in the world of prisoner in the situation of freedom deprivation is felt by him/her as difficult to support. This reality is had by him/her as difficult to control, having a negative emotional repercussion for the self, leading the citizen to the alienation of his/her existential flow in these circumstances. The reflections produced in this work take us to consider that the punishment practices on behalf of a normalization of the behavior, linked to the recovery idea, do not meet to the intention of the arrests, that come demonstrating to the inefficacy of the objectives shown in the law as well as the loss of the human being dignity

Utilização de uma coluna de flotação para remoção de compostos orgânicos da água de produção

Innovative technologies using surfactant materials have applicability in several industrial fields, including petroleum and gas areas. This study seeks to investigate the use of a surfactant derived from coconut oil (SCO saponified coconut oil) in the recovery process of organic compounds that are present in oily effluents from petroleum industry. For this end, experiments were accomplished in a column of small dimension objectifying to verify the influence of the surfactant SCO in the efficiency of oil removal. This way, they were prepared emulsions with amount it fastens of oil (50, 100, 200 and 400 ppm), being determined the great concentrations of surfactant for each one of them. Some rehearsals were still accomplished with produced water of the industry of the petroleum to compare the result with the one of the emulsions. According to the experiments, it was verified that an increase of the surfactant concentration does not implicate in a greater oil removal. The separation process use gaseous bubbles formed when a gas stream pass a liquid column, when low surfactant concentrations are used, it occurs the coalescence of the dispersed oil droplets and their transport to the top of the column, forming a new continuous phase. Such surfactants lead to a gas-liquid interface saturation, depending on the used surfactant concentration, affecting the flotation process and influencing in the removal capacity of the oily dispersed phase. A porous plate filter, with pore size varying from 40 to 250 mm, was placed at the base of the column to allow a hydrodynamic stable operation. During the experimental procedures, the operating volume of phase liquid was held constant and the rate of air flow varied in each experiment. The resulting experimental of the study hydrodynamic demonstrated what the capturing of the oil was influenced by diameter of the bubbles and air flow. With the increase flow of 300 about to 900 cm3.min-1, occurred an increase in the removal of oil phase of 44% about to 66% and the removal kinetic of oil was defined as a reaction of 1° order.

Um estudo algorítmico da programação da intervenção de sondas de produção

This work approaches the Scheduling Workover Rigs Problem (SWRP) to maintain the wells of an oil field, although difficult to resolve, is extremely important economical, technical and environmental. A mathematical formulation of this problem is presented, where an algorithmic approach was developed. The problem can be considered to find the best scheduling service to the wells by the workover rigs, taking into account the minimization of the composition related to the costs of the workover rigs and the total loss of oil suffered by the wells. This problem is similar to the Vehicle Routing Problem (VRP), which is classified as belonging to the NP-hard class. The goal of this research is to develop an algorithmic approach to solve the SWRP, using the fundamentals of metaheuristics like Memetic Algorithm and GRASP. Instances are generated for the tests to analyze the computational performance of the approaches mentioned above, using data that are close to reality. Thereafter, is performed a comparison of performance and quality of the results obtained by each one of techniques used

GeOntoQuery - um mecanismo de busca em bancos de dados geográficos baseado em ontologias

Geographic Information System (GIS) are computational tools used to capture, store, consult, manipulate, analyze and print geo-referenced data. A GIS is a multi-disciplinary system that can be used by different communities of users, each one having their own interest and knowledge. This way, different knowledge views about the same reality need to be combined, in such way to attend each community. This work presents a mechanism that allows different community users access the same geographic database without knowing its particular internal structure. We use geographic ontologies to support a common and shared understanding of a specific domain: the coral reefs. Using these ontologies' descriptions that represent the knowledge of the different communities, mechanisms are created to handle with such different concepts. We use equivalent classes mapping, and a semantic layer that interacts with the ontologies and the geographic database, and that gives to the user the answers about his/her queries, independently of the used terms

A gestão participativa do desenvolvimento rural sustentável: o PRONAF na Subzona de Touros/ RN

To investigate the possibilities and limits of mechanisms of participativa management in the scope of the Pronaf - National Program of Fortalecimento of Familiar Agriculture, and the corresponding effectiveness of its actions in the construction of alternatives of sustainable agricultural development in the Coast North of the State of the Great River of the North, more specifically, in the composed Subzona de Touros for the cities of Bulls, White Well, Pureness, Is Miguel of the Gostoso and Taipu. To carry through this analysis they had been carried through Research of Field with the objective to reconstitute the trajectory of the CMDRS, to trace the profile of the citizens that compose each one of the five advice, its expectations, interests and the level of participation of the local population

Códigos de sustentação da linguagem no cotidiano prisional do Rio Grande do Norte: penitenciária estadual de Parnamirim

The quarrel, in this study, tells about the language as social practical in the daily of Parnamirim State Prison that is integrated to the Rio Grande do Norte Penitentiary System - SISPERN, destined for men in fulfilment of penalty privative of freedom, in closed regimen. For the accomplishment of the research, the delimited objectives had been to analyze the language repertoires created in the prisional daily, trying to identify how it´s turned into distinct forms of resistance to the mechanisms of control in penitentiary system; to investigate which are the language repertoires created from the new sociability forms developed among prisoners and identify how the language repertoires are expressed in the daily prisional on relations/exercises of power not-institutionalized. In the methodological aspect, the study is in a qualitative boarding, that has as main instrument the interview. The inquiry was possible by means of using instruments for data collection, like as: the direct comment in the prisional daily duly registered as researcher´s ethnographical procedure, the analysis of interns´ cadastre handbooks and the application of half-structuralized interview, to the subjects of the research. The construction and understanding of the study object had been based on authors who argue on the arrest, as: Foucault, Goffman, Carvalho Filho and, in particular was searched the theorical referencial that approaches the language in a social and cultural perspective: Orlandi, Manfred, Bastos and Candiotto, amongst others. Beyond the normative endorsement of the Brazilian legislation, through the Law of Criminal Execution, of the Criminal Code and the Federal Constitution of the country and the legal apparatus in state scope. Still in the research methodological perspective, after the collection the data had been submitted to an analysis of the speech from Foucalt´s theory and in the Orlandi´s perspective, being also qualitative and quantitative. The results had evidenced that the social and juridical profile of the population in the site inquired is not different of others Brazilian prisons, composed for men, in its majority, with age band between 21 and 30 years old, prisoners for practicing crimes against the patrimony, against life, amongst others, and, in special, criminal recidivists. It evidenced, still, that the daily prisional of PSP is characterized for a sociocultural diversity expressed in the relations of power not institutionalized, that contributes for the formation and division of the groups, each one using a set of language codes/ repertoires sustentation. Therefore, the language, in the daily prisional, is one of the ways to understand the singularity of the sociability relations and as social practicing mediated by relations/exercises of power and antagonistic interests, in which each group aims first of all, their own interests. It represents the complexity of the social relations, in the prisional space, with diverse effects, in function of the situation and the moment. The language in the arrest, beyond the communication function, assumes and represents central element for the sociability human being, contributes for its changings and it´s configured as one of the resistance forms of prisoners against the controling, disciplining and monitoring mechanisms of penitentiary system