The pedunculopontine nucleus (PPN) is a brainstem structure involved in motor control, sleep and arousal. The boundaries of the PPN are defined by its cholinergic neurons, but it also contains GABAergic, glutamatergic and calcium-binding protein- positive neurons. To further understand the physiological roles of the PPN it is necessary to understand which neuronal subtypes are present in the PPN and how they are connected with other regions of the brain in normal and pathological conditions. In order to address these issues, the total numbers, distributions and neurochemical phenotypes of neurons, positive for the calcium-binding proteins calbindin and calretinin, were studied in the rat PPN. Sagittal, perfuse-fixed rat brain sections were double or triple-immunolabelled to reveal the cholinergic marker choline acetyltransferase (ChAT) with calbindin and/or calretinin. A stereological approach revealed that calbindin- and calretinin-positive neurons account for a large proportion of PPN neurons, but they rarely eo-express ChAT. A combination of immunolabelling for calbindin or calretinin with in situ hybridisation for GAD65/67 or VGluT2 mRNAs revealed that about one third of the calbindin- and calretinin-expressing neurons are GABAergic and preferentially located in the rostral PPN, whereas approximately two thirds are glutamatergic and principally located in the caudal PPN. Additionally, retrograde tracer injections in the subthalamic nucleus (STN) and the gigantocellular nucleus (GiN) showed that the majority of PPN neurons, projecting to one or both of these nuclei, were not cholinergic (70-90%). Less than 10% of STN-projecting neurons expressed calbindin or calretinin and 5% of the GiN-projecting neurons contained calretinin but none contained calbindin. Finally, the expression of the immediate early gene, Egrl, a marker of neuronal activation, was evaluated in STN- and GiN-projecting neurons of the PPN in control and 6-0HDA lesioned animals. No statistically significant differences, in the number of Egr l-positive neurons, were observed between control and 6- OHDA lesioned animals. These findings show that calbindin- and calretinin-positive neurons are abundant in the PPN, heterogeneously distributed and display a GABAergic or glutamatergic phenotype. Additionally, calbindin- and calretinin-positive neurons represent only a minority of the PPN neurons projecting to either the STN, GiN or both nuclei. Results also suggest that the hyperactivity seen in the PPN in the 6-0HDA model of Parkinson's disease may not necessarily be due to the neurons projecting to the STN and/or GiN. Overall, this thesis supports the notion that the PPN is composed of a rich diversity of neuronal cell-types, which are heterogeneously distributed along its rostro-caudal axis. The heterogeneous neurochemistry, connectivity and physiology of these neurons allow the PPN to influence a wide range of brain regions through a variety of pathways presumably underlying its various functional roles.

This thesis explored whether interpretations of the reinforcing effect of stimulation trains used in the self-administration of brain-stimulation (SABS) paradigm were artifacts of the reinforcement schedule chosen or whether it represented a genuine attempt by animals to maintain optimal levels of reward. Results demonstrate that stimulation trains used in SABS are reinforcing and that animals regulate pulse frequency to optimize the level of reward. The thesis then explored whether pedunculopontine tegmental nucleus (PPTg) lesions blocked the acquisition or maintenance of SABS, and the acquisition of eight-arm radial maze learning. Results showed that lesions confined to the PPTg block acquisition and maintenance of SABS, suggesting that the PPTg mediates the positive reinforcing effects of BSR. Further, PPTg lesions blocked win-shift and win-stay radial maze learning. However, results indicate that animals were not impaired in "shifting" or "staying" behavior. It is speculated that PPTg lesions block the reinforcing effects of food, which produce inefficient performance on both memory tasks.

The role of the pedunculopontine tegmental nucleus (PPTg) in motivation and cognitive functions is controversial. In order to clarify the involvement of this nucleus in learning, rats with N-methyl-D-Aspartate (NMDA) lesions to the PPTg were tested on the acquisition of a delayed non-matching to position task (DNMP) performed in a T-maze. Unlike sham-lesioned rats, animals with PPTg lesions did not learn the task. Analysis of the behavior displayed by the lesions animals, however, suggested that these rats suffered from elevated emotionality or arousal, rather than from learning deficits. This hypothesis was confirmed by demonstrating that the anxiolytic compound diazepam (1 mg/kg) normalized the performance of the PPTg-lesioned rats on the DNMP task and reduced the indices of anxiety displayed by animals with PPTg lesions when tested on the elevated plus maze. / These results suggested the possibility that the motivational impairments reported to be induced by PPTg lesions, could also be an artifact of lesion-induced elevation of anxiety or arousal. Thus, in order to verify this hypothesis, it was tested whether diazepam would modify the expression of conditioned place preference (CPP) to morphine and amphetamine in animals with NMDA-induced lesions to the PPTg. Diazepam reversed the effects of the lesion on a morphine CPP but not on an amphetamine CPP. A series of experiments, aimed at characterizing the effects of diazepam on morphine and amphetamine reinforcement in normal rats, showed that diazepam, either systemic or injected in the nucleus accumbens, blocks the reinforcing effects of amphetamine but has no effect on the reinforcing effects of morphine. These results suggest that impairments in CPP learning caused by PPTg lesions do not result from motivational deficits, but are caused by elevated emotionality or abnormal arousal.

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Orientador: Sergio Antonio De Bortoli / Resumo: No controle biológico, além dos microrganismos benéficos, devem ser também destacados parasitoides e predadores, sendo um bom exemplo deste último grupo, os crisopídeos (Neuroptera: Chrysopidae). O controle biológico aplicado, com a utilização de insetos predadores, como é o caso dos crisopídeos, está na dependência de alguns fatores, destacando-se dentre eles o conhecimento da biologia do inseto e a necessidade do domínio de uma técnica de criação eficiente e econômica em laboratório, que permita sua produção massal para subsequentes liberações em áreas de exploração agrícola. Já é de conhecimento que larvas de alguns crisopídeos são mantidas em laboratório, com boa eficiência, alimentadas com fontes de alimento obtidas de outras criações de insetos também mantidas em laboratório, dentre os quais podem ser destacados ovos de alguns lepidópteros. Ovos de Corcyra cephalonica (Stainton, 1866) (Lepidoptera: Pyralidae) são benéficos ao desenvolvimento de crisopídeos, produzindo adultos com alta viabilidade, porém há poucos relatos que estabelecem relação entre a espécie com Ceraeochrysa cincta (Schneider, 1851). Assim, este estudo teve por objetivo analisar o desempenho biológico de Ce. cincta alimentada com ovos de C. cephalonica, por três gerações, bem como fatores relacionados a técnicas de manipulação das diferentes fases deste predador na sua criação. Os ovos para os bioensaios com Ce. cincta foram coletados nas gaiolas de adultos com o auxílio de três instrumentos de corte,... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In biological control, in addition to beneficial microorganisms, parasitoids and predators should also be highlighted. A good example of this latter group is the crisopids (Neuroptera: Chrysopidae). The biological control applied, with the use of predatory insects, as in the case of the crisopids, is dependent on some factors, especially the knowledge of insect biology and the need to master an efficient and economical breeding technique in the laboratory, allowing its mass production for subsequent releases in farm areas. It is known that larvae of some chrysopids are maintained, with good efficiency, with food sources obtained from other insect creations also kept in the laboratory, among which lepidopteran eggs can be highlighted. Corcyra cephalonica (Stainton, 1866) (Lepidoptera: Pyralidae) has also been shown to favor the development of chrysopids, producing adults with high viability, not being many reports relating this species to Ceraeochrysa cincta (Schneider, 1851). Thus, this study aimed to analyze the biological performance of Ce. cincta being fed with eggs of C. cephalonica, per three generations as well as factors related to phase manipulation techniques of this predator in its rearing. The eggs of Ce. cincta were collected from adult cages with the aid of three cutting instruments, two types of scissors, one long and one conventional, with a thin tip, and an apparatus made from a stylus blade adapted to a shaver. Ten samples were collected for each of the three... (Complete abstract click electronic access below) / Mestre

Empresa Brasileira de Pesquisa AgropecuÃria / CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Na indÃstria de processamento de frutas, a polpa de caju à utilizada durante a entressafra do caju para a obtenÃÃo de diversos produtos, tais como o suco de caju. O consumo cada vez mais frequente de alimentos industrializados desperta o interesse na qualidade desses produtos que estÃo sendo consumidos e a influÃncia do processamento ainda à pouco estudada. Por isso a importÃncia em verificar a influÃncia desse processamento nos compostos benÃficos que estÃo presentes no suco de caju. Este trabalho visou quantificar o teor de fenÃlicos e avaliar a atividade antioxidante in vitro pelo mÃtodo ABTS no suco de caju processado. O suco foi obtido a partir dos pedÃnculos do clone de cajueiro CCP 76 em prensa tipo expeller. O tratamento tÃrmico foi realizado em escala de bancada com cinco faixas de temperaturas (55, 65, 75, 85 e 95ÂC) e quatro tempos (5, 15, 30 e 60 minutos). Imediatamente apÃs o tratamento tÃrmico, as amostras foram resfriadas em banho de gelo, e armazenadas em frascos plÃsticos para posterior anÃlise. Os Ãcidos fenÃlicos identificados foram 1-O-trans-cinamoil--D-glucopiranose, Ãcido cinÃmico e seus isÃmeros. Com relaÃÃo à atividade antioxidante, os resultados encontrados utilizando o radical ABTS, apresentaram valores TEAC de 7,11  0,08 para o extrato aquoso e 6,59  0,02 para o extrato metÃnÃlico (ÂM TROLOX/mL de suco). A partir desses resultados pode-se concluir que apesar da mudanÃa na concentraÃÃo dos compostos fenÃlicos durante o processamento, nÃo houve alteraÃÃo na atividade antioxidante do suco de caju em funÃÃo do tratamento tÃrmico. / In the fruit processing industry, the cashew pulp is used during the offseason to obtain various products such as cashew juice. The increasingly frequent consumption of processed foods arouses interest in the products that are being consumed and the influence of processing is still poorly studied. Hence the importance of verifying the influence of processing on the beneficial compounds present in cashew apple juice. This study aimed to quantify the amount of phenolics and evaluate the in vitro antioxidant activity by ABTS method in processed cashew juice. The juice was obtained from the peduncle of the cashew clone CCP 76 in an expeller type press. The thermal treatment was carried out in a bench scale at five temperature (55, 65, 75, 85 and 95  C) and four times (5, 15, 30 and 60 minutes). Immediately after the heat treatment, the samples were cooled in an ice bath and stored in plastic bottles for further analysis. The phenolics were identified as 1-O-trans-cinnamoyl--D-glucopyranose, cinnamic acid and isomers. With regard to antioxidant activity, the results using the ABTS radical, had TEAC values of 7.11  0.08 for the aqueous extract and 6.59  0.02 for the methanol extract (mM Trolox / mL of juice). From these results we can conclude that still of phenolic concentration changes during processing, there was no change on cashew juice antioxidant activity related to thermal treatment.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O uso de resÃduos agroindustriais puros ou modificados quimicamente para a remoÃÃo de metais em efluentes industriais aparece como uma alternativa promissora Ãs tecnologias existentes. Os agroresÃduos sÃo bastante estudados devido à sua abundÃncia, baixo custo, biodegradabilidade e a nÃo formaÃÃo de lodo. Neste trabalho utilizou-se o bagaÃo do pedÃnculo de caju (Anarcadium occidentale L.) como adsorvente para remoÃÃo de metais pesados de soluÃÃo aquosa e de efluente produzido por uma indÃstria de galvanoplastia localizada em Fortaleza (Cearà â Brasil). Foram estudados os sistemas de adsorÃÃo em batelada e coluna de leito fixo. Na etapa de batelada foram avaliados o tratamento quÃmico, o efeito do tamanho das partÃculas, o efeito da concentraÃÃo do adsorvente e o efeito do pH. Os resultados indicaram que o tratamento dado ao bagaÃo do pedÃnculo de caju com NaOH 0,1 mol.L-1/3 horas foi mais eficiente na remoÃÃo dos Ãons metÃlicos. Este material foi usado para o estudo de adsorÃÃo dos metais com as seguintes caracterÃsticas: faixa de tamanho de partÃculas de 20-59 mesh, concentraÃÃo de 50 g/L e pH 5,0. O estudo de cinÃtica indicou que o equilÃbrio de adsorÃÃo multielementar foi atingido em 60 minutos (pH 5,0) e segue cinÃtica de pseudo-segunda ordem. Foram utilizados os modelos de Langmuir e Freundlich para avaliar a capacidade de adsorÃÃo dos metais pesados pelo bagaÃo do pedÃnculo de caju. O modelo de Langmuir foi adequado para todos os Ãons metÃlicos em um sistema multielementar. Na segunda etapa da pesquisa foram realizados experimentos para otimizaÃÃo das condiÃÃes operacionais da coluna. Os resultados mostraram que uma vazÃo de 3,0 mL/min e uma altura de 12,0 cm de leito aumentaram o percentual de remoÃÃo para todos os metais tÃxicos estudados. As curvas de ruptura investigadas para um sistema multielementar indicaram que a ordem das capacidades de adsorÃÃo foi a seguinte: Pb2+ > Cu2+ > Ni2+ > Zn2+ > Cd2+. A aplicabilidade dos estudos de coluna numa amostra real, oriunda de empresa de galvanoplastia contendo Cu2+, Ni2+ e Zn2+, foi eficiente para remoÃÃo dos Ãons Ni2+ e Zn2+. A dessorÃÃo dos Ãons metÃlicos, retidos na coluna, foi eficiente usando HCl 1,0 mol/L como eluente. Os ensaios de reciclagem da coluna mostraram que o adsorvente pode ser utilizado apenas num ciclo. / Recently, the use of agroindustry residues (pure or chemically modified) for metal removal from industrial effluents offers a potential alternative to existing treatment technologies. The agrowastes are most studied because are readily available, cheap, biodegradable, sludge free and of low cost and land investment. In this work was used the cashew (Anacardium occidentale L.) peduncle bagasse as adsorbent for metal removal from aqueous solution and wastewater aqueous from electroplating industries local (Fortaleza, Ceara-Brazil). Adsorption systems in batch and fixed bed column were studied. Previously was investigated the chemical treatment, the particle size effect, adsorbent concentration and pH effect. The results showed that the treatment employed in the cashew peduncle bagasse with 0.1M NaOH for 3 h was the most efficient. The material characteristics: used on metal adsorption were: particle size ranged 20 â 59 mesh, adsorbent concentration (50g/L) at pH 5.0. The kinetic study indicated that the adsorption equilibrium multielementary were achieved in 60 minutes (pH 5.0) and followed a pseudo-second order model. The adsorption capacity of cashew peduncle bagasse for heavy metals had been evaluated using Langmuir and Freundlich adsorption isotherms. The Langmuirâs model gave best results for all metal ions in a multielementary system. In the second step of the work were carried out experiments to optimization of the column operational parameters. The results showed that for a flow rate of 3.0 mL/min and bed height of 12.0 cm there is an increased on the metal removal. The breakthrough curves for multielementary system indicated the order for adsorption capacity: Pb+2 > Cu+2 > Ni+2 > Zn+2 > Cd+2. The column parameters obtained with the synthetic effluents were applied in a real sample contained Cu+2, Ni+2 and Zn+2. It was found to be an efficient for the removal of Ni+2 and Zn+2. The metal desorption on the column was efficient using 1.0 M HCl as eluent. The tests of recycling of the column showed that the adsorbent can be used only for a cycle.

Três experimentos foram desenvolvidos em duas etapas, utilizando-se as dependências e instalações da Embrapa - Agroindústria Tropical, em Fortaleza, CE, com os seguintes objetivos: avaliar quanto aos aspectos físicos, físico-químicos, químicos e bioquímicos de pedúnculos de cajueiro anão precoce CCP-76 durante o desenvolvimento e maturação; caracterizar quantitativa e qualitativamente as alterações nos constituintes da parede celular do pedúnculo associadas aos processos de desenvolvimento e maturação, e verificar o efeito da aplicação pós-colheita de cálcio, em diferentes concentrações, sobre as características físicas, físico-químicas, químicas e bioquímicas dos pedúnculos submetidos a armazenamento refrigerado sob atmosfera modificada. Os cajus utilizados neste trabalho, foram obtidos na área comercial de clones de cajueiro anão precoce instalada na Empresa Mossoró Agro-industrial S.A. O clone selecionado para este estudo foi o CCP-76 por ser o mais cultivado para produção de pedúnculos para comercialização in natura. As colheitas foram feitas em junho e agosto de 1997 para os experimentos 1 e 2 respectivamente, e em novembro de 1998 para o experimento 3. Os cajus, depois de colhidos, foram imediatamente transportados para o laboratório, onde se deu início aos estudos conforme o experimento em questão. Para os experimentos 1 e 2, os cajus foram selecionados em sete estádios de desenvolvimento e maturação de acordo com a coloração externa do pedúnculo e da castanha: 1. verde com castanha verde; 2. verde com castanha madura e seca; 3. verde claro; 4. Início de coloração amarela; 5. Amarelo com início de cor laranja; 6. laranja claro e 7. laranja escuro. No primeiro experimento, foram realizadas análises físicas, físico-químicas, químicas e bioquímicas dos pedúnculos: peso total (castanha + pedúnculo) peso do pedúnculo e o da castanha, diâmetros e comprimento, firmeza, clorofilas, carotenóides, antocianinas, pH, sólidos solúveis, acidez, vitamina C, cálcio, amido, açúcares, compostos fenólicos, pectinas, atividade das enzimas poligalacturonase, pectinametilesterase, polifenoloxidase e peroxidase. No segundo experimento, a partir de material de parede celular extraído, foram realizadas as seguintes análises: celulose, hemicelulose, uronídeos totais, grau de esterificação de pectinas e cálcio ligado. No terceiro experimento, os cajus foram colhidos no estádio de maturação comercial (estádio 7) e submetidos a seleção, préresfriamento, desinfecção em água clorada, secagem ao ar, nova seleção e tratamento com CaCI2 a 0,5%, 1% e 2% com espalhante adesivo, sendo que a testemunha foi apenas em água contendo o espalhante. Após secagem ao ar, os cajus foram acondicionados em bandejas, as quais foram envolvidas em PVC armazenadas em câmara com temperatura de 5°C ± 1°C e umidade relativa de 88% ± 3%. A cada 5 dias, por um período de 25 dias, foram tomadas amostras para execução das seguintes análises: perda de peso, firmeza, senescência, antocianinas, pectinas, compostos fenólicos, vitamina C, pH, sólidos solúveis, acidez, açúcares solúveis, cálcio, atividades de pectinamestilesterase e poligalacturonase. A maior relação sólidos solúveis/acidez foi atingida no estádio 7, indicando que este é o ideal para a colheita de pedúnculos ocasião em que alcança sua máxima qualidade para o consumo. Os pedúnculos de caju apresentaram amaciamento acentuado durante o desenvolvimento e maturação, observando-se paralelamente redução no teor de pectina total e aumento na percentagem de solubilização. A firmeza dos pedúnculos apresentou correlação significativa e inversa com a solubilidade das pectinas e com a atividade das enzimas PME e PG. A solubilidade da pectina, por sua vez, mostrou correlação significativa e positiva com a atividade de ambas as enzimas, que se correlacionaram entre si positivamente. O ambiente refrigerado aliado a atmosfera modificada, pode ser usado com sucesso no retardamento do processo de senescência do pedúnculo de caju, uma vez que aumenta substancialmente sua vida pós-colheita sem causar injúrias fisiológicas por até 20 dias. Pedúnculos tratados com cálcio foram mais firmes do que os não tratados, sendo que a partir de 0,5% não se observou um aumento do efeito. Doses crescentes de cálcio proporcionaram teores médios de cálcio total, solúvel e insolúvel superiores aos da tetemunha. O tratamento com cálcio não inibiu a atividade da pectinametilesterase. Porém foi decisivo na atividade de poligalacturonase, uma vez que os pedúnculos não tratados obtiveram valores médios de atividade dessa enzima superiores aos tratados. / Three experiments were carried out in two stages, using the EMBRAPA (Tropical and Agricultural Industry) - plants and annexes in Fortaleza, CE aiming the following: to evaluate the physícal, physicochemical, chemical and biochemical characteristícs of cashew apples of the clone CCP-76 during maturation and developing stages; to characterize qualitative and quantitative changes in the cell wall contents of the cashew apples due to developing and maturation processes; and to verify the post harvest application effect of calcium under different concentrations on cashew apples. The physical, physicochemical, chemical and biochemical characteristics of cashew apples stored under refrigeration and modified atmosphere were studied. The cashew apples used for this work were obtained from the clone experimental areas of early dwarf cashew trees of Mossoró Agricultural Company S.A. The selected clone for this study was the CPP-76 due to be recommended for commercial planting in function of its production aspects and cashew apple quality for consumption in natura. The harvests were done in June and August 1997 for the 1st and 2nd experiments, respectively and in November 1998 for the 3rd experíment. The cashew apples, after being harvested were transported immediately to the laboratories, for the starting of the due experiment. For the experiments 1 and 2, the cashew apples were selected in seven developing and maturation stages according to the cashew apple size and skin externai color as the following: 1- green cashew apple and nut; 2- green cashew apple with ripen dry nut; 3- light green cashew apple, 4- yellowish, 5- onset of the orange color, 6- light orange color, 7- dark orange color. For the first experiment the following physical, physicochemical, chemical and biochemical analyses were realized: total weight (cashew apple + nut), nut weights, cashew apple weights, diameters and lengths, firmness, chlorophylls, carotenoids, anthocyanins, pH, total soluble solids, titratable acidity, vitamin C, calcium, starch, sugars, phenolic compounds, pectins, polygalacturonase, pectinmethylesterase, polyphenoloxidade e peroxidase activities. In the second experiment, the following analyses on the cell wall material extracted, were realized: cellulose, hemicellulose, total uronides, pectin esterification degree and bound calcium. For the third experiment, the cashew apples were harvested on the commercial maturation stage and submitted to a selection, pre-cooling, chlorinated water washing, air drying, new selection and treatment with CaCI2 0.5%, 1% and 2% with thickener being the testimony, water and thickener only. After air drying, the cashew apples were placed on trays, involved with PVC film and stored at 5°C ± 1°C and relative humidity 88% ± 3%. Samples were taken for analyses at each 5 days during a 25 day period and the following analyses carried out: weight loss, firmness, senescence, anthocyanins, pectins, tannins, vitamin C, pH, total soluble solids, titratable acidity, calcium, soluble sugars, polygalacturonase, pectinmethylesterase activities. The major ratio soluble solids/acidity was reached in the stage 7, indicating that stage as the ideal for cashew apple harvesting, occasion that the edible quality peak is reached. The cashew apples showed a sharp softness during the development and maturation in parallel to a decrease in total pectin levei and increase in soluble pectin/total pectin ratio. There was an inverse and significative correlation between cashew apples firmness and pectin solubility, PME and PG activities. On the other hand, pectin solubility showed a significative and positive correlation with the activity of both enzymes, which correlated, positively to each other. The refrigeration added to the modified atmosphere may be used with success to delay cashew apple senescence process once they increase substantially its post harvest shelf life without causing physiological damage up to 20 days of storage. The cashew apples treated with calcium showed better firmness than those without treatment did, although, under calcium concentration up to 0,5% that effect was not observed anymore. Increased calcium dosages gave mean levei of total, soluble and insoluble calcium higher than the testimony. The calcium treatment did not inhibit the pectinmethylesterase activity. However, calcium was decisive for polygalacturonase activity, once the cashew apples that did not receive calcium treatment, obtained mean values for that enzyme activity higher than those treated.