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

Análise de grafos aplicada a relatos de sonhos: ferramenta diagnóstica objetiva e diferencial para psicose esquizofrênica e bipolar

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Análise da expressão imuno-histoquímica da ciclofilina A, EMMPRIN e MMP-7 na doença periodontal

Periodontal disease is an infectious disease resulting from the immunoinflammatory response of the host to microorganisms present in the dental biofilm which causes tissue destruction. The objective of this study was to evaluate the immunohistochemical expression of cyclophilin A (CYPA), extracellular matrix metalloproteinase inducer (EMMPRIN) and matrix metalloproteinase 7 (MMP-7) in human specimens of clinically healthy gingiva (n=32), biofilm-induced gingivitis (n=28), and chronic periodontitis (n=30). Immunopositivity for CYPA, EMMPRIN and MMP-7 differed significantly between the three groups, with higher percentages of staining in chronic periodontitis specimens, followed by chronic gingivitis and healthy gingiva specimens (p < 0.001). Immunoexpression of CYPA and MMP-7 was higher in the intense inflammatory infiltrate observed mainly in cases of periodontitis. Analysis of possible correlations between the immunoexpression of EMMPRIN, MMP-7 and CYPA and between the expression of these proteins and clinical parameters (probing depth and clinical attachment loss) showed a positive correlation of CYPA expression with MMP-7 (r = 0.831; p < 0.001) and EMMPRIN (r = 0.289; p = 0.006). In addition, there was a significant positive correlation between probing depth and expression of MMP-7 (r = 0.726; p < 0.001), EMMPRIN (r = 0.345; p = 0.001), and CYPA (r = 0.803; p < 0.001). These results suggest that CYPA, EMMPRIN and MMP-7 are associated with the pathogenesis and progression of periodontal disease

Consumo e digestibilidade aparente de dietas com níveis crescentes do subproduto do caju em ovinos

The present study was conducted to evaluate the intake and digestibility of diets containing increasing levels of byproduct of cashew in sheep. The animals were distributed in a completely randomized design were evaluated in four levels (0%, 20%, 40%, 60%) by product of cashew with four replicates, making up 16 observations. The indicator used was the Purified Lignin and Enriched - LIPE ®. The scorer was orally administered directly into the mouth of the animals, in the form of capsules 250mg/animal/dia for a period of two days and five days of adaptation samples being the same supplied with the aid of a hose polyethylene and a device allowing the release of the capsule in the esophagus of sheep. With the estimate made by the indicator LIPE was observed a reduction for DM, OM, CP, NDF, EE, NFC and MM along the inclusion of byproduct of cashew. The results of nutrient digestibility were not satisfactory with the inclusion of byproduct of cashew, reducing linearly with the inclusion of the diets. The use of increasing levels of byproduct of cashew in the diets of sheep did not provide satisfactory results, it is not feasible to use the animals studied in this experiment

Respostas morfogênicas e estruturais de gramíneas tropicais em regime de corte no nordeste brasileiro

The production of forage grasses is directly related to the morphogenesis. The knowledge of the morphogenetic and structural variables of forage plants is important for determining appropriate conditions of grazing livestock to ensure efficient and sustainable. Thus the objective of this study was to evaluate morphogenetic and structural responses of three genera of grasses, Brachiaria, Panicum and Cenchrus in a cutting regime. The experimental design was randomized blocks with three replications and six treatments. After each section were evaluated for forage production, appearance and elongation rates of leaves and stem, phyllochron, final leaf length, number of living leaves, leaf lifespan, leaf senescence rate, tiller density and tiller dynamics. On forage yield the highest values were obtained in cultivars Xaraes, Piata and Massai. The tiller density was higher for cv Massai. It is concluded that the cultivars of Panicum and Brachiaria had a higher tillering dynamics in increasing the turnover rate of tissues that are indicators of forage production, assuming that the cultivars of these genera are predisposed to use forage in the Northeast

Sistema de produção misto de caprino na região central do estado do Rio Grande do Norte

This study aimed to evaluate the performance and profitability of Canindé goats kept under semi-intensive management in mix farming system. 84 animals were used, being 81 does e three bucks.. The means of fertility rate at parturition, prolificacy and percentages of pregnant, kid survival and reproductive rate were compared by Tukey test at significance level of 5.0%. The kid weights from birth to weaning were submitted to variance analysis considering sex and birth type and the means were compared by Tukey test at 5.0% of significance. The percentages of fertility rate by ultrassographic were 91.36 and parturition 74.84. The prolificacy was 1.53. Independently of sex the mortality rate of kids from birth to weaning was 12.3. The average weights of offspring were 2.11 and 1.92 kg and 6.74 and 6.84 kg for male and female offspring at birth and weaning, in this order. The average weights of kids, meaning PN, P28, P56, P63, in the first season of kidding were 2.11 ± 0.46 kg, 3.63 ± 1.04 kg, 5.96 ± 158 kg and 6.74 ± 1.79 kg. There was significant differences (P<0,05) only to birth weight of kids sons of buck 135. However, the fertility at parturition for first and second breeding seasons (BS) were 87.65 and 62.96, respectively (P<0.05). The prolificacy was 1.58 and 1.47 to first and second BS in this order. The percentages of offspring survival from birth to weaning, to first and second BS were 92.86 and 80.00, respectively. From the first BS the reproduction rate was 1.28 while in the second was 0.74. After weaning the duration of lactation was 104 days to first breeding parturition and 38 days in the second one. The total milk production was 853.50 kg. The kids weight gain was influenced by season of year, wet and dry and buck (P<0.05). Were concluded that the seasons of year when breeding season and parturition occur strongly affected the productive performance and profitability of production system

Levantamento exploratório da composição e frações proteicas do leite bovino em propriedades leiteras

The direct relationship with the processing, industrial yield and price of milk, the importance of the centesimal composition of milk and their protein fractions has increased for both the dairy industry and for the producers, so it is important to identify the possible causes on variation of these components. The aim of this study was to evaluate the effect of the seasons, order of calving and lactation phase on milk composition and protein fractions in dairy farms in the Agreste and Leste meso-regions, of Rio Grande do Norte State (RN). Milk samples were taken in seven milk farms directly from the bulk tanks except for one farm where samples were taken from higher milking cows producers and heifers. The experiment was done in seven properties with milk samples taken directly from the bulk tanks and in one property were collected from higher production cows and heifers. Composition analyses were carried out for both experiments and sensorial trial was performed for the second experiment only. The data results were submitted to analysis of variance (ANOVA) and Tukey test performed for 5% significance level. In the first experiment was observed response of the seasons, dry and rainy season; while in the second, the effect of the order of calving and lactation phase had influence on milk composition and protein fractions. There was significant response (p < 0.05) for season effects but due to farms. The rainy season had higher averages in the protein, lactose, total solids and nonfat dry extract. Multiparous cows had higher CCS, NUL and PCAS milk contents; while, first calving heifers showed higher average for the sensory attribute, color of milk. The final third of lactation was responsible for the elevation of the levels of almost all milk components and their protein fractions

Parâmetros parasitológicos e desempenho produtivo de ovinos naturalmente infectados com nematoides gastrintestinais mantidos em diferentes pastagens tropicais

The control of gastrointestinal nematodes of small ruminants is usually done with anthelmintics. However, due to the emergence of ever-increasing parasite resistance to these drugs, looking up an alternative control parasites. One of this is sought in pasture management, as these are the sources of animals` infection by L3 infective larvae of helminths. The aim of this study was to evaluate the performance of sheep to natural infections by gastrointestinal nematodes grazing Panicum maximum cv. Massai and cv. Aruana, and Brachiaria brizantha cv. Piatã and cv. Marandu. The work was conducted from May to August-2011 with 48 male sheeps SRD versus Santa Inês breed. The animals were naturally infected with gastrointestinal nematodes, and maintained in four different cultivars of tropical forage grasses, naturally contaminated with eggs and larvae of gastrointestinal nematodes. Each week the animals were phenotypically characterized by parasitological examinations (eggs per gram of feces - EPG, and feces culture), hematological (packed cell volume PCV, and blood eosinophil count) method to evaluate the Famacha© colorof ocular mucosa, and the measures of body condition score and weight. In pastures was made the recovery of infective larvae in order to determine the quantity of L3 present in the pasture. The experimental design was a randomized completed block with two replications and before the entry of animals in the paddocks, they have been wormed. The experiment was ended when the animals reached 32.0 kg liveweight, and then were slaughtered and autopsies performed for the recovery and identify parasites of the abomasum, small intestine and large intestine. Results were subjected to analysis of variance, t test and Tukey`s test. The animals kept on pastures of Marandu grass had lower EPG counts, higher percentage of packed cell volume and higher average weight; those who remained in the Piatã pasture had lower eosinophil counts per microliter of blood. About the Famacha©, the highest prevalence was Famacha 2, and the body condition score ranged between 2 and 3. The results of feces cultures and recovery of larvae on pastures showed the presence of larvae of Trichostrongylus sp., and at the necropsy too. This way, it was concluded that the grass cultivars influences the sheep parasite load; the Famacha, together with EPG and packed cell volume are important indicators for use in controlling gastrointestinal nematode infections. The Trichostrongylus sp. was the most prevalent parasite in sheep during the rainy season

Estimação de parâmetros genéticos para resistência à infecção por IMNV em camarões Litopenaeus vannamei por meio de análise de sobrevivência

The main specie of marine shrimp raised at Brazil and in the world is Litopenaeus vannamei, which had arrived in Brazil in the `80s. However, the entry of infectious myonecrosis virus (IMNV), causing the infectious myonecrosis disease in marine shrimps, brought economic losses to the national shrimp farming, with up to 70% of mortality in the shrimp production. In this way, the objective was to evaluate the survival of shrimps Litopenaeus vannamei infected with IMNV using the non parametric estimator of Kaplan-Meier and a model of frailty for grouped data. It were conducted three tests of viral challenges lasting 20 days each, at different periods of the year, keeping the parameters of pH, temperature, oxygen and ammonia monitored daily. It was evaluated 60 full-sib families of L. vannamei infected by IMNV in each viral challenge. The confirmation of the infection by IMNV was performed using the technique of PCR in real time through Sybr Green dye. Using the Kaplan-Meier estimator it was possible to detect significant differences (p <0.0001) between the survival curves of families and tanks and also in the joint analysis between viral challenges. It were estimated in each challenge, genetic parameters such as genetic value of family, it`s respective rate risk (frailty), and heritability in the logarithmic scale through the frailty model for grouped data. The heritability estimates were respectively 0.59; 0.36; and 0.59 in the viral challenges 1; 2; and 3, and it was also possible to identify families that have lower and higher rates of risk for the disease. These results can be used for selecting families more resistant to the IMNV infection and to include characteristic of disease resistance in L. vannamei into the genetic improvement programs

Valor nutritivo de fenos de moringa (moringa oleifera lam) com diferentes idades de corte

This study was conducted to evaluate the consumption and digestibility of dry matter (DM), organic matter (OM), crude protein (CP), ether extract (EE), total carbohydrates (TC), non-fiber carbohydrates (NFC) and neutral detergent fiber (NDF) in sheep fed hay moringa (Moringa oleifera Lam) obtained with four cutting ages (28, 35, 42 and 49 days). We used 20 females Morada Nova breed, with 20kg of live weight, distributed in a completely randomized design and maintained in metabolism cages. There was a negative linear effect of age of cutting on DM intake, with an estimated maximum consumption of 0.67 kg / day for the hay produced at 28 days of cutting. It was also observed linear behavior, with an estimated maximum consumption 172g/dia, 0.36 kg / day; 18g/dia, at 35, 42 and 49 days old, for CP, OM and EE, respectively. For NDF, quadratic effect was found with advancing maturity of the plant. The apparent digestibility of DM, CP, OM and TC linearly decreased with advancing age of cutting hay Moringa oleifera. It was concluded that the hay Moringa oleifera showed better nutritional value after 28 days of cutting

Composição ictiofaunística e estratégias reprodutivas de quatro espécies de peixes nativos da Bacia Hidrográfica Piranhas - Assu, Rio Grande do Norte

This study focuses on the fish fauna composition and reproductive strategies of four native fish species in relation to environmental variables of the Piranhas-Assu hydrographic basin of the Caatinga biome, Rio Grande do Norte, Brazil. Fish and environmental data were collected monthly during the period September 2008 to February 2010, in the Marechal Dutra reservoir and River Acauã a tributary of Piranhas- Assu. The fish were measured, weighed and dissected, and the gonads were removed, weighed and examined macroscopically for identification and determination of gonad maturation stages. The results of this work generated seven articles. The first article is about the fish fauna composition of the Piranhas-Assu hydrographic basin, Rio Grande do Norte. The 602 fish samples captured were distributed in four orders (Characiformes, Perciformes, Siluriformes and Synbranchiformes), 11 families and 22 species, of which 17 are endemic to the Caatinga ecoregion. The order Characiformes was more representative followed by Perciformes, Siluriformes and Synbranchiformes. The second article is about the length-weight relationship and growth of seven native fish species Crenicichla menezesi, Cichlasoma orientale, Triportheus angulatus, Psectrogaster rhomboides, Pimelodella gracilis, Prochilodus brevis and Leporinus piau from a semiarid Brazilian reservoir. The third article is about the reproductive aspects of Crenicichla menezesi. Males were larger, heavier and with a slight predominance as compared to females. Four stages of gonadal development were characterized, being immature, maturing, mature and spent. The females reached sexual maturity earlier than males, with an average fecundity of 398 oocytes per batch. The spawning was partial with a long reproductive period. The fourth article deals with the reproductive strategy of Leporinus piau a neotropical freshwater fish in semi-arid region of Brazil. The population of L. piau (n = 211) showed a slight predominance of males (55%), with larger and heavier females. The males matured earlier than the females. This species presented total spawning, with an average fecundity of 55,000 mature oocytes. Rainfall and concentration of dissolved oxygen acted as influential factors during the spawning season. L. piau shows a seasonal reproductive strategy. The fifth article is related to the morphometric-meristic characteristics and reproductive aspects of freshwater sardine, Triportheus angulatus from River Acauã of the Caatinga biome. There was a predominance of larger females, reaching first maturation before males. There was total spawning during the rainy period of the region. The sixth article reports on the reproductive strategy of Psectrogaster rhomboides. The sex ratio was 1M: 1F, with negative allometric growth. Males reached sexual maturity earlier than females Females and males showed four stages of gonadal development and spawned during the rainy season. The fecundity was low and this species presented total spawning. The seventh article reports on the dynamics of territorial behavior of Crenicichla menezesi. Ten agonistic behaviors displayed by the males were observed: frontal and lateral threat, chasing, circular chasing, perpendicular, lateral and mouth attacks, escape, parallel positioning and stationary. The formation of four social groups was observed among males: without interaction; interaction with submission and escape; with frontal and lateral agonistic interactions. In social interactions between males and females, it was observed that the larger males interacted more among themselves and with the larger females. The large male established its territory and the two small males along with the small female were excluded from the other groups. These studies clarified the fish fauna composition and reproductive strategies of four native species Crenicichla menezesi, Leporinus piau, Triportheus angulatus and Psectrogaster rhomboides of the Piranhas- Assu hydrographic basin of the Caatinga biome, Rio Grande do Norte, Brazil.

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

Fatores que influenciam a cooperação em humanos

Human cooperation is fundamentally affected by reciprocal exchange, but it is also remarkably common on the context of large and symbolically marked in-groups, which promote cooperation through the feeling of belonging to a group. In this thesis, two empirical articles were produced in order to investigate how human cooperation is affected by factors such as reciprocity, in-group behavior, in-group markers and gender. We investigated this subject through the administration of online games consisting of token donations, on which the subjects faced virtual players controlled by the experiment. We found that cooperative behavior is strongly influenced by reciprocity, and it is also affected by the in-group behavior, observed on the context of the social variables place of birth, ethnicity, and religions, once all of them acted as in-group markers. The subjects´ in-group behavior was enhanced when they played with generous in-group opponents, but weakened when their in-group opponents were non-generous. It was also found that cooperation is not affected by gender, but men and women cooperated in different ways under the influence of reciprocity and in-group behavior. Women are much more reciprocal on their cooperative behavior and men are less willing to cooperate with outgroupers, even when they act generously. The overall results contribute to a better understanding of the adaptive value of cooperation, reciprocity and in-group behavior on the solution of important challenges through the human evolutionary history

Características sociodemográficas, hábitos de sono, estado cognitivo e funcional após acidente vascular cerebral

ABSTRACT Introduction: The cerebrovascular accident (CVA) is an important cause of neurological impairment. Few data about the factors associated with morbidity of cerebrovascular accident are found in Brazil. Objectives: Evaluate sociodemographic characteristics, sleep habits, cognitive and functional status of patients with cerebrovascular accident. Methods: The patients evaluated through questionnaire Step 1 to survey the sociodemographic characteristics and Modified Rankin Scale for functional assessment. The neurological degree was evaluated by the National Institutes of Health Stroke Scale (NIHSS), the sleep Habits questionnaire for sleep and cognitive status by the Mini-Examination of the Mental State (MEMS). The data were analyzed using the chi-square test to determine differences in proportions of variables and linear regression analysis. Results: 305 patients were evaluated and the larger number of subjects was between 50 and 69 years (40%), most patients had no formal education (40.3%) and had ischemic type of cerebrovascular accident (72.5%). In the analysis of the functionality it was found that most patients had moderate impairment (55.1%). The results of the sleep habits showed that 63,6% of patients had one more person in the bedroom,12,3% complained about too much noise in the 11 room and 35% of too much light. From these patients 5,8% were smokers, 7,8% and 70,1% drank coffee drinkers, 28,6% had difficulty in initiate to sleep and woke up 37,6% in the middle of the night. Were showed complaints about nightmares (11%), feeling of suffocation (37,7%) and 35% felt very sleepy during the day. In addition, 95% were unemployed, 80,5% did not perform physical activities and 95,4% did not perform mental activities. The cognitive screening conducted a determined association of cognitive status with age and education level and neurological status. Conclusion: The study showed a high frequency of cases of cerebrovascular accident with functional dependence in a moderate degree, identified that many patients do not follow hygienic measures of sleep and found that the assessment of cognitive deficits must take into consideration the age, educational level and degree of neurological patients. We suggest the need for programs of assistance to victims of cerebrovascular accident patients, with a multidimensional approach including the rehabilitation team, the role of sleep medicine and Neuropsychology, so that patients have access to a more appropriate functional rehabilitation, develop a lifestyle that ensures a good sleep quality and are evaluated and rehabilitated with regard to cognitive impairment

Taxa de disparo e ativação de assembléias no hipocampo de ratos durante o processamento de uma memória aversiva

Hebb postulated that memory could be stored thanks to the synchronous activity of many neurons, building a neural assembly. Knowing of the importance of the hippocampal structure to the formation of new explicit memories, we used electrophysiological recording of multiple neurons to access the relevance of rate coding from neural firing rates in comparison to the temporal coding of neural assemblies activity in the consolidation of an aversive memory in rats. Animals were trained at the discriminative avoidance task using a modified elevated plus-maze. During experimental sessions, slow wave sleep periods (SWS) were recorded. Our results show an increase in the identified neural assemblies activity during post-training SWS, but not for the neural firing rate. In summary, we demonstrate that for this particular task, the relevant information needed for a proper memory consolidation lies within the temporal patters of synchronized neural activity, not in its firing rate