Análise fitoquímica e avaliação das atividades anti-inflamatória, antipeçonhenta e citotóxica de extratos aquosos de Aspidosperma pyrifolium e Ipomoea asarifolia

Envenomation caused by venomous animals, mainly scorpions and snakes, are a serious matter of public health. Tityus serrulatus is considered the most venomous scorpion in South America because of the high level of toxicity of its venom. It is responsible for causing serious accidents, mainly with kids. The species Bothrops jararaca is a serpent that has in its venom a complex mixture of enzyme, peptides and other molecules. The toxins of the venom of B. jararaca induce local and systemic inflammatory responses. The treatment chosen to serious cases of envenomation is the intravenous administration of the specific antivenom. However, the treatment is not always accessible to those residents in rural areas, so that they use medicinal plant extracts as the treatment. In this context, aqueous extracts, fractions and isolated compounds of Aspidosperma pyrifolium (pereiro) and Ipomoea asarifolia (salsa, salsa-brava), used in popular medicine, were studied in this research to evaluate the anti-inflammatory activity in the peritonitis models induced by carrageenan and peritonitis induced by the venom of the T. serrulatus (VTs), and in the local oedema model and inflammatory infiltrate induced by the venom of the B. jararaca, administrated intravenously. The results of the assays of cytotoxicity, using the MTT, showed that the aqueous extracts from the plant species presented low toxicity to the cells that came from the fibroblast of the mouse embryo (3T3).The chemical analysis of the extracts by High Performance Liquid Chromatography revealed the presence of the rutin flavonoid, in A. pyrifoliu, and rutin, clorogenic acid and caffeic acid, in I. asarifolia. Concerning the pharmacological evaluation, the results showed that the pre-treatment using aqueous extracts and fractions reduced the total leukocyte migration to the abdominal cavity in the peritonitis model caused by the carrageenan and in the peritonitis model induced by the T. serulatus venom. Yet, these groups presented anti-oedematous activity, in the local oedema model caused by the venom of the B. jararaca, and reduced the inflammatory infiltrate to the muscle. The serum (anti-arachnid and anti-bothropic) specific to each venom acted inhibiting the inflammatory action of the venoms and were used as control. The compounds identified in the extracts were also tested and, similar to the plant extracts, showed meaningful anti-inflammatory effects, in the tested doses. Thus, these results are indicating the potential anti-inflammatory activity of the plants studied. This is the first research that evaluated the possible biological effects of the A. pyrifolium and I. asarifolia, showing the biological potential that these species have.

Recuperação e purificação do antígeno 503 de Leishmania i. chagasi expresso em E. coli e remoção de endotoxina utilizando adsorção em leito expandido

The growing interest and applications of biotechnology products have increased the development of new processes for recovery and purification of proteins. The expanded bed adsorption (EBA) has emerged as a promising technique for this purpose. It combines into one operation the steps of clarification, concentration and purification of the target molecule. Hence, the method reduces the time and the cost of operation. In this context, this thesis aim was to evaluate the recovery and purification of 503 antigen of Leishmania i. chagasi expressed in E. coli M15 and endotoxin removal by EBA. In the first step of this study, batch experiments were carried out using two experimental designs to define the optimal adsorption and elution conditions of 503 antigen onto Streamline chelating resin. For adsorption assays, using expanded bed, it was used a column of 2.6 cm in diameter by 30.0 cm in height coupled to a peristaltic pump. In the second step of study, the removal of endotoxin during antigen recovery process was evaluated employing the non-ionic surfactant Triton X-114 in the washing step ALE. In the third step, we sought developing a mathematical model able to predict the 503 antigen breakthrough curves in expanded mode. The experimental design results to adsorption showed the pH 8.0 and the NaCl concentration of 2.4 M as the optimum adsorption condition. In the second design, the only significant factor for elution was the concentration of imidazole, which was taken at 600 mM. The adsorption isotherm of the 503 antigen showed a good fit to the Langmuir model (R = 0.98) and values for qmax (maximum adsorption capacity) and Kd (equilibrium constant) estimated were 1.95 mg/g and 0.34 mg/mL, respectively. Purification tests directly from unclarified feedstock showed a recovery of 59.2% of the target protein and a purification factor of 6.0. The addition of the non-ionic surfactant Triton X-114 to the washing step of EBA led to high levels (> 99%) of LPS removal initially present in the samples for all conditions tested. The mathematical model obtained to describe the 503 antigen breakthrough curves in Streamline Chelanting resin in expanded mode showed a good fit for both parameter estimation and validation steps. The validated model was used to optimize the efficiencies, achieving maximum values of the process and of the column efficiencies of 89.2% and 75.9%, respectively. Therefore, EBA is an efficient alternative for the recovery of the target protein and removal of endotoxin from an E. coli unclarified feedstock in just one step.

Mapeamento cromossômico de DNAs repetitivos com fins biotecnológicos em peixes marinhos e interesse comercial - Rachycentridae e Lutjanidae

The Rachycentron canadum species, commonly known as beijupirá or cobia is the only representative of Rachycentridae family which has been increasingly used in marine fish farming, in intensive cultivation. As advantageous features it has easy adaptation, prolific behavior, early growth in captivity and high commercial value. Additionally, specie of Lutjanidae family (Lutjanus synagris, Lutjanus jocu, Lutjanus analis, Lutjanus alexandrei and Ocyurus chrysurus) represents an important fisheries resource in all areas of its occurrence. In Brazil, the commercial exploitation of Lutjanidae which begun in the 60's and 80's, already has showed a decline in catch volumes. This fact suggests that the snappers must have a conservative management. Despite the economic potential, little is known about the genetic and cytogenetic characteristics of these species, especially with respect to repetitive DNA analysis, which represents the major part of the eukaryotes genome, playing important evolutionary roles in the fish genome. Cytogenetic data is increasingly being used in population studies and biotechnological purposes in fishes. The cytogenetical analyzes were performed using classical methods such as Giemsa staining, C-banding and Ag-NORs, fluorochromes base-specific staining (DAPI and MM) and physical mapping of repetitive sequences among which, telomeric sequences, transposons (Tol2), retrotransposons (Rex1 and Rex3), repetitive DNA (microsatellites and Cot-1) and transcriptionally active regions of the 18S and 5S ribosomal genes and histone (H3 and H2BA) by in situ hybridization with fluorescent probes (FISH). The chromosomal patterns obtained contributed to the organization of repetitive sequences in the genome of the species, as well as karyotypical differentiation. Unusual patterns of histone sequences expansion depict the first occurrence in marine fishes. The obtained data provided subsides to the genetic knowledge of the important fisheries resource represented by the species here analyzed, seeking the marine pisciculture improvement.

Efeito da Olmesartana na resposta inflamatória em modelo de mucosite intestinal em ratos

Intestinal Mucositis is inflammation and/or ulceration of mucosa of the gastrointestinal tract caused by anticancer therapies. Histologically, villous atrophy, damage to enterocytes and infiltration of inflammatory cells. Methotrexate (MTX) is a compound that depletes dihydrofolate pools and is widely used in the treatment of leukemia and other malignancies. The aim of this study was to evaluate the effect of Olmesartan (OLM), an angiotensin II receptor antagonist, on an Intestinal Mucositis Model (IMM) induced by MTX in Wistar rats. IMM was induced via intraperitoneal (i.p.) administration of MTX (7 mg/kg) for three consecutive days. The animals were pretreated with oral OLM at 0.5, 1 or 5 mg/kg or with vehicle 30 min prior to exposure to MTX, for three days. Small intestinal (duodenum, jejunum and ileum) homogenates were assayed for levels of the IL-1β, IL-10 and TNF-α cytokines, malondialdehyde and myeloperoxidase activity. Additionally, immunohistochemical analyses of MMP-2, MMP-9, COX-2, RANK/RANKL and SOCS-1 and confocal microscopy analysis of SOCS-1 expression were performed. Treatment with MTX+OLM (5 mg/kg) resulted in a reduction of mucosal inflammatory infiltration, ulcerations, vasodilatation and hemorrhagic areas (p<0.05) as well as reduced concentrations of MPO (p<0.001) and the pro-inflammatory cytokines IL-1β and TNF-α (p<0.01), and increase antiinflammatory cytosine IL-10 (p,0.05). Additionally, the combined treatment reduced expression of MMP-2, MMP-9, COX-2, RANK and RANKL (p<0.05) and increased cytoplasmic expression of SOCS-1 (p<0.05). Our findings confirm the involvement of OLM in reducing the inflammatory response through increased immunosuppressive signaling in an IMM. We also suggest that the beneficial effect of Olmesartan treatment is specifically exerted during the damage through blocking inflammatory cytosines.

A influência da relevância social no viés de grupo

Behaviors found in every culture, general human tendencies, are knew in Evolutionary Psychology as evolved psychological mechanisms. Those behaviors date back the Environment of Evolutionary Adaptedness, and a well know example of such behavior is the group bias (or intergroup bias). This bias consists of recognizing members of your own group and favor them, while disregarding or even harming outsiders. This behavior was and still is extensively studies, among the most important conclusions about this phenomenon is the Minimal Groups Paradigm, in which it was discovered that the group bias could trigger even when the groupings were done in following very arbitrary criteria. In the current study, our goal was to test if the participants, when playing an economic game, would behave in a similar fashion under a minimal group situation and real groups, with social meaning. With this in mind we made two experimental conditions, a Low Social Meaning one (LSM) where the groups were represented by letters (H, B, O and Y) in which participants would be ramdomly assorted to each group; and the High Social Meaning condition (HSM) in which religion was used as a group marker, containing the two most dominating religious groups in Brazil, catholic and evangelic, another group containing all the other affiliations e the fourth and last group representing atheists and agnostics. The ratio of donations in-group/out-group was roughly the same across both conditions. However, the amount of wafers donated to ingroup was significantly bigger in the HSM condition. By verifying which aspects of the individual best predicted the observed group bias, we discovered that the in-group Entitativity perception as well as the Group Identification were the most relevant variables, however, only in the HSM condition. Simultaneously, by verifying the generosity, biased or not, we observed that the agreeableness personality factor was the only variable able to predict it, and only in the LSM condition. We conclude that our generosity, or the lack of it, is for most part defined by our personality, the Agreeableness factor in particular. But this very generosity can be biased by the social meaning of the involved groups and that, if the social meaning is big enough, even people who, thanks to their personality, normally wouldn’t show generosity, are able to do so when the receiver is an in-group member.

Relação do polimorfismo BDNF val66met e níveis periféricos de BDNF com a doença de Parkinson e sua sintomatologia

Neurodegenerative diseases are frequently studied due to the increasing number of cases associated with the populational ageing and to the impact on the conditions on the quality of life. Parkinson’s disease (DP) is the second most frequent neurodegenerative disease. Despite the fact that its etiology is not completely understood, it is known that DP is caused by environmental and genetic factors. Thus, the investigation of etiologic factors and mechanisms responsible for the changes that lead to DP may help early diagnostic and prevention. A possible association between DP and the common polymorphism of Brain Derived Neurotrophic Factor (BDNF) G196A (Val66Met) has been suggested by different studies with contrasting results. For this reason, the aim of this study is to investigate if the BDNF Val66Met polymorphism is related to susceptibility to DP in a cohort of Brazilian patients. Additionaly, we verify if the presence of the polymorphism implies in alterations in the BDNF whole blood concentrations, as well as variations in symptomatology. The sample comprised Brazilian patients accompanied by the neurology service of the Onofre Lopes University Hospital (HUOL) and healthy controls (CTRL). The motor aspects of DP were evaluated by Hoehn e Yahr Scale (HY), Unified Parkinson’s Disease Rating Scale (UPDRS) and Schwab & England Scale (SE). For the evaluation of non-motor symptoms were used the following instruments: Frontal Assessment Battery (BAF), Mini-Mental State Examination (MEEM), Beck Depression Inventory (IDB) and the Beck Anxiety Inventory (IAB). Blood samples were collected for BDNF Val66Met polymorphism genotyping and BDNF whole blood measurement. As expected, DP patients performed worse in motor, cognitive and emotional battery of questionnaires. Alleles distribution between DP and CTRL was not significantly different, but the A/G genotype was significantly associated with a protector factor for DP. In contrast, the G/G genotype was significantly associated with depression and anxiety development in DP patients. However, BDNF concentrations were not different between genotypes or groups. This is the first study of genetic association of this polymorphism with DP in Brazilian subjects and the first one that associate A/G genotype with protection against DP.

Estabelecimento, reconhecimento e defesa territorial em Stegastes fuscus

Os estudos da territorialidade e de comportamentos associados a ela favorecem o entendimento da maneira como as interações ecológicas afetam a composição de espécies e a dinâmica de uma comunidade. No presente estudo tivemos como objetivo geral investigar o comportamento de Stegastes fuscus, um peixe-donzela territorialista, em ambiente natural e em cativeiro, com foco na capacidade de localização territorial, reconhecimento e defesa de uma área estabelecida. Para tanto subdividimos o trabalho em 3 capítulos. O primeiro teve como foco o estudo da espécie em ambiente natural objetivando estimar a área do território ocupado e os padrões comportamentais da por ela expressos. Sendo encontrado que a área média ocupada por S. fuscus foi de 274 cm2 e os comportamentos mais observados foram: vigilância, ingestão de alimento, tempo no abrigo/toca e displays agressivos. O segundo capítulo teve como alvo investigar a capacidade de localização espacial da espécie mediada por pistas visuais. Os resultados demonstraram que S. fuscus apresenta marcante aprendizagem condicionada e possibilidade de existência de orientação espacial na espécie. O terceiro capítulo teve como objetivo avaliar a influência da residência prévia estabelecida e do reconhecimento de coespecíficos nos resultados de confrontos agonísticos. Os resultados apontaram a residência como fator prioritário na dinâmica das disputas agonísticas e que aspectos relacionados à familiaridade como relevantes e destacam-se mais quando não existe um território previamente estabelecido. Diante disso nossos resultados podem favorecer o entendimento da dinâmica estrutural da comunidade na qual S. fuscus esta inserida, sendo isto significativo tendo em vista a importância ecológica da espécie para o ecossistema.

Caracterização citoarquitetônica e por imunoistoquímica para tirosina-hidroxilase da substância negra, área tegmentar ventral e zona retrorubral do Sagui (Callithrix jacchus)

It is known that the catecholamine group is constituted by dopamine, noradrenaline and adrenaline, in which the synthesis is regulated by an enzyme named tyrosine hydroxylase. Thus, 3-hydroxytyramine/dopamine (DA) is a precursor of the noradrenaline and adrenaline synthesis and acts as a neurotransmitter in the central nervous system. The three main nuclei, named the retrorubral field (A8 group), the substantia nigra pars compacta (A9 group) and the ventral tegmental area (A10 group), are arranged in the die-mesencephalic portion and are involved in three complexes circuitries - the mesostriatal, mesolimbic and mesocortical pathways. These pathways are related to behavioral manifestations, motricity, learning, reward and pathologies such as Parkinson’s Disease and Schizophrenia. Thus, the aim of this study was to perform de morphological analysis of the A8, A9 and A10 nuclei of the common marmoset (Callithrix jacchus). The marmoset is a neotropical primate, whose morphological and functional characteristics supports the suitability of use of this animal in biomedical research. Coronal sections of the marmoset brain were submitted to cytoarchitectonic characterization and TH-immunohistochemistry. Based on the morphology of the neurons, it was possible to subdivide the A10 group in seven regions: interfascicular nucleus, raphe rostral linear nucleus and raphe caudal linear nucleus, in the middle line; paranigral and parainterfascicular nucleus, in the middle zone; rostral portion of the ventral tegmental area nucleus and parabrachial pigmented nucleus, located in the dorsolateral portion of the mesencephalic tegmentum. A9 group was divided into four regions: substantia nigra compacta dorsal and ventral tiers; substantia nigra compacta lateral and medial clusters. No subdivisions were founded into A8 group. These results revealed that A8, A9 and A10 are phylogenetically conserved between species, but it’s necessary to expand the studies about this compartmentalization, investigating its occurrence in other primate species or investigating its functional relevance.

Efeito anticonvulsivante de frações isoladas da peçonha da formiga Dinoponera quadriceps (Formicidae: Ponerinae)

A epilepsia é uma patologia crônica do sistema nervoso central que afeta cerca de 65 milhões de indivíduos no mundo. Aproximadamente 30% desses indivíduos desenvolvem crises convulsivas que persistem apesar do tratamento monitorado com drogas antiepiléptica s. Assim, há uma evidente necessidade do desenvolvimento de novos fármacos antiepilépticos e as peçonhas podem ser uma excelente fonte de modelos. Nesse contexto, enquanto já vários estudos sobre peçonhas de serpentes, escorpiões e aranhas, pouco se sabe s obre as peçonhas de formigas. Estudos prévios do nosso laboratório demonstraram que a peçonha desnaturada da formiga Dinoponera quadríceps protegeu camundongos de crises convulsivas e morte induzidas por bicuculina (BIC). Nesse contexto, o objetivo desse t rabalho foi investigar o potencial anticonvulsivante de frações isoladas da peçonha de D. quadríceps em crises convulsivas induzidas pela BIC, bem como uma análise dos efeitos dessas frações no comportamento natural dos camundongos no campo aberto. Os anim ais foram divididos em grupos, os quais receberam injeções (1 mg/ml i.c.v.) de seis frações distintas e tiveram seu comportamento geral observado no campo aberto durante 3 0 min. No segundo experimento, o s animais receberam as mesmas frações 20 min antes da administração de bicuculina (10 mg/ml). Em seguida, foi analisado o comportamento motor convulsivo desses animais durante 30 minutos no campo aberto. No primeiro experimento, não foram observadas alterações comportamentais. Já no segundo experimento, a ad ministração prévia de DqTx1, DqTx3, DqTx4 e DqTx6 aumentou a latência para o desenvolvimento de crises tônico - clônicas. Além disso, todas as frações, exceto DqTx5, aumentaram a latência para a morte dos animais. Ainda, os melhores resultados foram obtidos com a fração DqTx6, que protegeu 62,5% dos animais testados contra o desenvolvimento de crises tônico - clônicas e 100% dos animais contra a morte.

Citoarquitetura e imunoistoquímica para tirosinahidroxilase dos núcleos dopaminérgicos mesencefálicos do morcego (Artibeus planirostris)

The 3-Hidroxytyramine/dopamine (DA) is a monoamine of catecholamine family and isthe precursor substance synthesis of noradrenaline and adrenaline, having the enzymeTyrosine Hydroxylase (TH) as this regulatory process. In addition, the DA has theability to act as a neurotransmitter in the Central Nervous System - SNC, being themain neurotransmitter of brain nuclei, namely of A8 to A16. The nuclei of the midbrainthat express DA are the Retrorubral Field (RRF, A8), the Substantia Nigra parsCompacta (SNc, A9) and the Ventral Tegmental Area (VTA, A10). Such nuclei areinvolved in complex three circuitry that are the mesostriatal, mesolimbic andmesocotical and are directly related with several behavioral manifestations as motricitycontrol, reward signaling in behavioral learning, motivation and pathologicalconditions, such as Parkinson's Disease and schizophrenia. Interestingly, many of themorphological bases of these neural disturbance remain unknown. Considering therelevance of mesencephalic dopaminergic nuclei, the aim of this research is tocharacterize morphologically the dopaminergic nuclei (clusters A8, A9 and A10) of themidbrain of the bat (Artibeus planirostris). The Artibeus planirostris is a common bat inRio Grande do Norte. Ten animals were used in this research. The animals wereanesthetized, perfused, and the brain was removed from the skull. After dehydration insucrose, the brain was underwent microtomy. Saggital and coronal sections wereobtained and collected in six separate series. The series were Nissl-stained to identifythe cytoarchitectonic boundaries and the other series were subjected toimmunohistochemistry for TH. After cytoarchitectonic analysis and TH+ cellsidentification was possible to establish the anatomical boundaries of the nuclei, as wellas the subdivisions of three of the midbrain dopaminergic nuclei. The SNc is the mostrostral nucleus observed in the midbrain and is identified throughout the rostrocaudalextension of the midbrain. The VTA neurons were seen immediately caudal to the SNcappearance. The RRF neurons were observed just in the caudal levels of the midbrain.The SNc in Artibeus planirostris shows a particular feature, the tail of the SNc. The tailhave been described just in two other studied species. The present work shows aparticular variation in the organizational morphology of the SNc in the artibeus andcontribute to understand the phylogenetic routes by which the dopaminergic system hasevolved.

Efeito do ácido ascórbico sobre a espermatogênese de camundongos com distrofia muscular de Duchenne

The objective was to evaluate spermatogenesis alterations caused by DMD and the effect of the treatment using ascorbic acid in preventing those injuries. Twenty four mice were used, 12 from the C57BL/10 lineage (non-dystrophic) and 12 from the C57BL/10Mdx (dystrophic). The sample was divided in six groups containing 4 animals each, as: C30 = 30 days control; D30 = Dystrophic with 30 days; C60 = 60 days control; D60 = Distrophic with 60 days; CS60 = 60 days control supplemented with ascorbic acid and DS60 = Dystrophic with 60 days supplemented with ascorbic acid. The ascorbic acid supplementation was given in the water, 0,005 mg/day. After euthanasia, the testicles (right and left) were collected, weighted and cross sectioned. The material was fixed in the Karnovsky solution for 24 hours, included in resin for histological studies (morphological and morphometric analyzes) submitted to ultrastructural analysis and immunohistochemistry for caspase-3. There was a significant increase in the tunica propria percentage in D30 compared to C30 and D60. The ultrastructural analysis showed mitochondrial apoptosis evidence of Sertoli cells that can reduce sperm efficiency in CS60 and DS60. A higher volume density of apoptotic cells postivas to Caspase-3 in C30 and D30 versus DS60 compared to CS60. There was severe hypertrophy of the Leydig cells between D30 and D60. However, with supplementation was observed reversal of this change in DS60. The ultrastructure of Leydig cells to early presence of lipid vesicles was observed in the group pre-pubertal dystrophic (D30). Thus, the DMD affect the organization of the seminiferous tubules and intertubule, however, the ascorbic acid supplementation used for the treatment of DMD has been just enough to reduce the hypertrophy of the Leydig cells.

Influência dos odores de gato e de cobra no comportamento defensivo e expressão de fos do sistema hipotalâmico de defesa de camundongo

Studies using neuronal tract-tracer in rat have shown that the anterior hypothalamic nucleus, dorsomedial division of the ventromedial nucleus of the hypothalamus and dorsal premammillary nucleus are highly connected. When the rat is exposed to predator or its odor these nuclei have shown a expression of Fos and their lesion reduces defensive behavior against predator. This set of nuclei was named the Hypothalamic Defense System. However, little is known about the response of this system to the odor of different predators or its role in mice. In this work, we exposed Swiss mice to two different predators odor (cat and snake) to verify the Fos expression in the Hypothalamic Defense System, as well as the defensive behaviors displayed. The analysis showed that the mice exposure to cat odor had an increased expression of Fos protein compared to control, while those exposed to snake odor showed no rise in Fos expression, which was corroborated by the behavioral data. Our results indicate that this distinct circuit in mice seems to act differentially to odorous stimuli of different predators, causing distinct behavioral responses of mice and that the odor of snake seems not to be perceived by Swiss mice as a threatening stimulus.

When wolves cry: long distance calling by wild maned wolves

The maned wolf (Chrysocyon brachyurus Illiger 1815) is the biggest canid in South America and it is considered a “near threatened” species by IUCN. Because of its nocturnal, territorial and solitary habits, there are still many understudied aspects of their behavior in natural environments, including acoustic communication. In its vocal repertoire, the wolf presents a longdistance call named “roar-bark” which, according to literature, functions for spacing maintenance between individuals and/or communication between members of the reproductive pair inside the territory. In this context, this study aimed: 1) to compare four methods for detecting maned wolf’s roar-barks in recordings made in a natural environment, in order to elect the most efficient one for our project; 2) to understand the night emission pattern of these vocalizations, verifying possible weather and moon phases influences in roarbark’s emission rates; and 3) to test Passive Acoustic Monitoring as a tool to identify the presence of maned wolves in a natural environment. The study area was the Serra da Canastra National Park (Minas Gerais, Brazil), where autonomous recorders were used for sound acquisition, recording all night (from 06pm to 06am) during five days in December/2013 and every day from April to July/2014. Roar-barks’ detection methods were tested and compared regarding time needed to analyze files, number of false positives and number of correctly identified calls. The mixed method (XBAT + manual) was the most efficient one, finding 100% of vocalizations in almost half of the time the manual method did, being chosen for our data analysis. By studying roarbarks’ temporal variation we verified that the wolves vocalize more in the early hours of the evening, suggesting an important social function for those calls at the beginning of its period of most intense activity. Average wind speed negatively influenced vocalization rate, which may indicate lower sound reception of recorders or a change in behavioral patterns of wolves in high speed wind conditions. A better understanding of seasonal variation of maned wolves’ vocal activity is required, but our study already shows that it is possible to detect behavioral patterns of wild animals only by sound, validating PAM as a tool in this species’ conservation.

Organização nuclear do sistema serotonérgico no encéfalo de morcegos Artibeus planirostris

Serotonin or 5-hydroxytryptamine (5-HT) is a substance found in many tissues of the body, including the nervous system acting as a neurotransmitter. Within the neuro-axis, the location of the majority of the 5-HT neurons is superimposed with raphe nuclei of the brain stem, in the median line or its vicinity, so that neuronal 5-HT can be considered a marker of the raphe nuclei. Serotonergic neurons are projected to almost all areas of the brain. Studies show the participation of serotonin in regulating the temperature, feeding behavior, sexual behavior, biological rhythms, sleep, locomotor function, learning, among others. The anatomy of these groups has been revised in many species, including mouse, rabbit, cat and primates, but never before in a bat species from South America. This study aimed to characterize the serotonergic clusters in the brain of the bat Artibeus planirostris through immunohistochemistry for serotonin. Seven adult bat males of Artibeus planirostris species (Microchiroptera, Mammalia) were used in this study. The animals were anesthetized, transcardially perfused and their brains were removed. Coronal sections of the frozen brain of bats were obtained in sliding microtome and subjected to immunohistochemistry for 5-HT. Delimit the caudal linear (CLi), dorsal (DR), median (MnR), paramedian (PMnR), pontine (PNR), magnus (MgR), pallidus (RPA) and obscurus (ROb) raphe nucleus, in addition to the groups B9 and rostral and caudal ventrolateral (RVL/CVL). The serotonergic groups of this kind of cheiroptera present morphology and cytoarchitecture relatively similar to that described in rodents and primates, confirming the phylogenetic stability of these cell clusters.

Construção de bibliotecas de cDNAs subtraídos a partir das sementes de Mamona submetidas à estresse hídrico e análise in silico de transcritos potencialmente expressos sob condições de estresse: Metalotioneina, proteína reprimida por auxina de 12.5 kDa e Glutelina tipo-A3

Estresses ambientais abióticos são fatores que causam respostas ao nível molecular, fisiológico e morfológico em plantas, dependendo também de sua intensidade e duração. É visto que algumas espécies apresentam tolerância a condições estressantes e ao mesmo tempo são fontes naturais de matéria prima para indústria. Nesse contexto encontra-se a mamona (Ricinus comunnis L.), principal fonte de óleo de rícino valorizado por suas aplicações farmacêuticas e principalmente industriais, vem sendo usada como cultura em regiões onde a disponibilidade de água é reduzida, usada como fonte de renda para agricultura da região nordeste brasileira. Visto que pouco se sabe sobre as respostas moleculares que levam essa planta a tolerar regiões secas e como as sementes, principais foco de interesse, respondem a essa escassez, nesse trabalho foram construídas duas bibliotecas de cDNAs, onde a partir de uma abordagem subtrativa, continham RNAs diferencialmente expressos em sementes de plantas mamona submetidas ao estresse hídrico durante 5 dias (biblioteca L7), e a outra RNAs diferencialmente expressos em sementes controle (biblioteca L5). A biblioteca L7 apresentou a maior variedade de transcritos com um total de 182. A maior parte das funções estabelecidas pelo sistema Gene Ontology - GO, foram direcionadas aos “Processos Metabólicos” (526), em segundo “Respostas a estímulos” (57), o terceiro termo mais abundante foram referentes a “Desenvolvimento”(26). Já na biblioteca L5, foram encontrados 91 transcritos, com maior parte de suas funções referentes a “Processos Metabólicos”(413), em segundo “Respostas a estímulos” (8) e em terceiro Regulação (6). Alguns dos transcritos da biblioteca L7 foram escolhidos para análise por repetirem-se mais de 3x e não aparecerem na biblioteca L5, o que indica uma possível regulação positiva sobre estresse. As análises sobre Metalotioneína (4x), mostraram que a sequência de proteica apresentava os domínios conservados que a caracterizava como tipo II, onde são encontrados dois domínios funcionais ricos em cisteína com posições altamente conservadas, desempenhando a função de ligar-se a metais pesados, correlacionadas assim como a atividade de eliminação EROs e defesa contra o estresse oxidativo, além de apresentar homologia com a sequência de Bruguiera gymnorhiza, uma planta de mangue adaptada a ambientes salinos. Analisamos também os transcritos da referente a proteína AUXIN-REPRESSED 12.5 KDA (3x), apontada como sendo reprimida pelo hormônio auxina e associada ao processo de dormência da semente, é descrito em uma família gênica onde vários membros pertencem as vias de resposta ao estresse. Por último, analisamos a proteína GLUTELIN TYPE-A 3 (5x), uma importante proteína de armazenamento com caráter hidrofílico, possivelmente direcionada para o vacúolo. Em nosso trabalho foi possível observar um aumento de transcritos em relação a subtração controle, possivelmente reflexo do aumento do metabolismo da semente, tanto para resposta defensiva ao estresse hídrico quanto para o amadurecimento rápido da semente onde foram observados transcritos referentes a resposta oxidativa, controle hormonal, proteínas de reserva e produção de óleo.