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Crack cocaine is a highly addictive and potent stimulant drug. Animal studies have shown that the cholinergic system plays a role in neurotoxicity induced by cocaine or its active metabolites inhalation. Behavioral alterations associated with crack cocaine use include hyperactivity, depressed mood, and decreased seizure threshold. Here we evaluate the acetylcholinesterase (AChE) and reactive oxygen species (ROS) activity, behavioral profile, and the threshold for epileptic seizures in rats that received intrahippocampal pilocarpine (H-PILO) followed by exposure to crack cocaine (H-PILO + CRACK). Animals exposed to H-PILO + CRACK demonstrated increased severity and frequency of limbic seizures. The AChE activity was reduced in the groups exposed to crack cocaine alone (CRACK) and H-PILO + CRACK, whereas levels of ROS remained unchanged. In addition, crack cocaine exposure increased vertical locomotor activity, without changing water and sucrose intake. Short-term memory consolidation remained unchanged after H-PILO, H-PILO + CRACK, and CRACK administration. Overall, our data suggest that crack cocaine inhalation reduced the threshold for epileptic seizures in rats submitted to low doses of pilocarpine through the inhibition of AChE. Taken together, our findings can be useful in the development of effective strategies for preventing and treating the harmful effects of cocaine and crack cocaine on the central nervous system.

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The population growth trend in recent decades has resulted in continuing efforts to guarantee food security in which leguminous plants, such as the common bean (Phaseolus vulgaris L.), play a particularly important role as they are relatively cheap and have high nutritional value. To meet this demand for food, the main target for genetic improvement programs is to increase productivity, which is a complex quantitative trait influenced by many component traits. This research aims to identify Quantitative Trait Nucleotides (QTNs) associated with productivity and its components using multi-locus genome-wide association studies. Ten morpho-agronomic traits [plant height (PH), first pod insertion height (FPIH), number of nodules (NN), pod length (PL), total number of pods per plant (NPP), number of locules per pod (LP), number of seeds per pod (SP), total seed weight per plant (TSW), 100-seed weight (W100), and grain yield (YLD)] were evaluated in four environments for 178 Mesoamerican common bean domesticated accessions belonging to the Brazilian Diversity Panel. In order to identify stable QTNs, only those identified by multiple methods (mrMLM, FASTmrMLM, pLARmEB, and ISIS EM-BLASSO) or in multiple environments were selected. Among the identified QTNs, 64 were detected at least thrice by different methods or in different environments, and 39 showed significant phenotypic differences between their corresponding alleles. The alleles that positively increased the corresponding traits, except PH (for which lower values are desired), were considered favorable alleles. The most influenced trait by the accumulation of favorable alleles was PH, showing a 51.7% reduction, while NN, TSW, YLD, FPIH, and NPP increased between 18 and 34%. Identifying QTNs in several environments (four environments and overall adjusted mean) and by multiple methods reinforces the reliability of the associations obtained and the importance of conducting these studies in multiple environments. Using these QTNs through molecular techniques for genetic improvement, such as marker-assisted selection or genomic selection, can be a strategy to increase common bean production.

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KEY MESSAGE: QTNs significantly associated to nine mineral content in grains of common bean were identified. The accumulation of favorable alleles was associated with a gradually increasing nutrient content in the grain. Biofortification is one of the strategies developed to address malnutrition in developing countries, the aim of which is to improve the nutritional content of crops. The common bean (Phaseolus vulgaris L.), a staple food in several African and Latin American countries, has excellent nutritional attributes and is considered a strong candidate for biofortification. The objective of this study was to identify genomic regions associated with nutritional content in common bean grains using 178 Mesoamerican accessions belonging to a Brazilian Diversity Panel (BDP) and 25,011 good-quality single nucleotide polymorphisms. The BDP was phenotyped in three environments for nine nutrients (phosphorus, potassium, calcium, magnesium, copper, manganese, sulfur, zinc, and iron) using four genome-wide association multi-locus methods. To obtain more accurate results, only quantitative trait nucleotides (QTNs) that showed repeatability (i.e., those detected at least twice using different methods or environments) were considered. Forty-eight QTNs detected for the nine minerals showed repeatability and were considered reliable. Pleiotropic QTNs and overlapping genomic regions surrounding the QTNs were identified, demonstrating the possible association between the deposition mechanisms of different nutrients in grains. The accumulation of favorable alleles in the same accession was associated with a gradually increasing nutrient content in the grain. The BDP proved to be a valuable source for association studies. The investigation of different methods and environments showed the reliability of markers associated with minerals. The loci identified in this study will potentially contribute to the improvement of Mesoamerican common beans, particularly carioca and black beans, the main groups consumed in Brazil.

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The common bean (Phaseolus vulgaris L.) is of great importance to the food and nutritional security of many populations, and exploitation of the crop's genetic diversity is essential for the success of breeding programs. Thus, the aim of the present study was to evaluate the genetic diversity of 215 common bean accessions, which included cultivars, obsolete cultivars, improved lines, and landraces using morpho-agronomic and biochemical traits, and amplified fragment length polymorphism markers (AFLP). Genetic parameters, box plots, Pearson's correlation analysis, and Ward's hierarchical clustering were used to analyze the data. The Jaccard similarity coefficient and neighbour-joining clustering method were used for molecular analysis. A wide variability among the accessions was observed for morpho-agronomic and biochemical traits. Selective accuracy (Ac) and broad-sense heritability (h2) values were high to intermediate for all traits, except seed yield. Ward's hierarchical clustering analysis generated six groups. AFLP analysis also revealed significant differences among the accessions. There was no correlation between the differences based on genetic markers and those based on morpho-agronomic and biochemical data, which indicates that both datasets are important for elucidating the differences among accessions. The results of the present study indicate great genetic diversity among the evaluated accessions.

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Brazil is the world's largest producer of common bean. Knowledge of the genetic diversity and relatedness of accessions adapted to Brazilian conditions is of great importance for the conservation of germplasm and for directing breeding programs aimed at the development of new cultivars. In this context, the objective of this study was to analyze the genetic diversity, population structure, and linkage disequilibrium (LD) of a diversity panel consisting of 219 common bean accessions, most of which belonging to the Mesoamerican gene pool. Genotyping by sequencing (GBS) of these accessions allowed the identification of 49,817 SNPs with minor allele frequency > 0.05. Of these, 17,149 and 12,876 were exclusive to the Mesoamerican and Andean pools, respectively, and 11,805 SNPs could differentiate the two gene pools. Further the separation according to the gene pool, bayesian analysis of the population structure showed a subdivision of the Mesoamerican accessions based on the origin and color of the seed tegument. LD analysis revealed the occurrence of long linkage blocks and low LD decay with physical distance between SNPs (LD half decay in 249 kb, corrected for population structure and relatedness). The GBS technique could effectively characterize the Brazilian common bean germplasms, and the diversity panel used in this study may be of great use in future genome-wide association studies.

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Crack users suffer the effects of cocaine present in the drug and the action of other active compounds from its pyrolysis. An emergent fact is an increase in the number of pregnant crack cocaine users. Studies suggest that crack cocaine and its metabolites cross the placenta, promoting premature birth, fever, irritability, sweating, and seizures in the early months of life. In children, the effects of crack cocaine have been associated with cognitive deficits, difficulty in verbalization, aggressiveness, and depression, besides enhancing the susceptibility to epileptic seizures, including status epilepticus (SE) in adulthood. Therefore, we investigated the effect of maternal exposure to smoke crack cocaine on several behavioral parameters in the offspring during adulthood. A series of behavioral tests and intrahippocampal pilocarpine (H-PILO) microinjection at sub-convulsive and convulsive doses in a rat model demonstrated that exposure to crack cocaine during the embryonic period leads to anxiogenic-like behavior and long-term memory impairment in both genders and promotes depressive-like behavior in the female. Besides, crack cocaine offspring exposed to a sub-convulsive H-PILO dose showed higher susceptibility to SE, increased seizure frequency, and neurodegeneration, while animals that received a convulsive dose of H-PILO displayed no alteration in SE severity. Taken together, our data suggest that crack cocaine exposure during the gestational period leads to an increased predilection for anxiety and depression, long-term memory deficits, and reduction in the threshold for developing epileptic seizures associated with neuronal death, which predispose crack cocaine babies to develop neuropsychological disorders.

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Mineral deficiency is worldwide one of the major problems associated with human health, and biofortification through breeding is considered an important strategy to improve the nutritional content of staple food in countries that face this problem. The assessment of genetic variability for seed nutrient contents is a first step in the development of a biofortified crop. From the germplasm bank IDR-IAPAR-EMATER, a set of 1,512 common bean accessions, consisting of local and commercial varieties and improved lines, was analyzed. High variability among the accessions was observed for all evaluated nutrient contents (P, K, Ca, Mg, Cu, Zn, Mn, Fe and S and protein). In the mean, the contents of the carioca and black market groups (Mesoamerican gene pool), were around 7% higher for the minerals Ca, Cu, Mn and Fe and between 2-4% higher for P, K, Mg and Zn than in the other groups with Mesoamerican and Andean common bean. Few differences were observed among the Mesoamerican accessions that belong to the carioca and black commercial groups. Wide variability was observed among the evaluated genotypes, and the concentrations of the best accessions exceeded the overall mean by 14-28%. Due to the high variability in the evaluated accessions, these results may contribute to the selection of promising parents for the establishment of mating blocks. The nutritional contents of many of the improved lines evaluated in this study were higher than those of the commercial cultivars, indicating the possibility of developing new biofortified cultivars.

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Some 1,3-diarylureas and 1-((1,4-trans)-4-aryloxycyclohexyl)-3-arylureas (cHAUs) activate heme-regulated kinase causing protein synthesis inhibition via phosphorylation of the eukaryotic translation initiation factor 2 (eIF2) in mammalian cancer cells. To evaluate if these agents have potential to inhibit trypanosome multiplication by also affecting the phosphorylation of eIF2 alpha subunit (eIF2α), we tested 25 analogs of 1,3-diarylureas and cHAUs against Trypanosoma cruzi, the agent of Chagas disease. One of them (I-17) presented selectivity close to 10-fold against the insect replicative forms and also inhibited the multiplication of T. cruzi inside mammalian cells with an EC50 of 1-3 µM and a selectivity of 17-fold. I-17 also prevented replication of African trypanosomes (Trypanosoma brucei bloodstream and procyclic forms) at similar doses. It caused changes in the T. cruzi morphology, arrested parasite cell cycle in G1 phase, and promoted phosphorylation of eIF2α with a robust decrease in ribosome association with mRNA. The activity against T. brucei also implicates eIF2α phosphorylation, as replacement of WT-eIF2α with a non-phosphorylatable eIF2α, or knocking down eIF2 protein kinase-3 by RNAi increased resistance to I-17. Therefore, we demonstrate that eIF2α phosphorylation can be engaged to develop trypanosome-static agents in general, and particularly by interfering with activity of eIF2 kinases.

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Some 1,3-diarylureas and 1-((1,4-trans)-4-aryloxycyclohexyl)-3-arylureas (cHAUs) activate heme-regulated kinase causing protein synthesis inhibition via phosphorylation of the eukaryotic translation initiation factor 2 (eIF2) in mammalian cancer cells. To evaluate if these agents have potential to inhibit trypanosome multiplication by also affecting the phosphorylation of eIF2 alpha subunit (eIF2 alpha), we tested 25 analogs of 1,3-diarylureas and cHAUs against Trypanosoma cruzi, the agent of Chagas disease. One of them (I-17) presented selectivity close to 10-fold against the insect replicative forms and also inhibited the multiplication of T. cruzi inside mammalian cells with an EC50 of 1-3 mu M and a selectivity of 17-fold. I-17 also prevented replication of African trypanosomes (Trypanosoma brucei bloodstream and procyclic forms) at similar doses. It caused changes in the T. cruzi morphology, arrested parasite cell cycle in G1 phase, and promoted phosphorylation of eIF2 alpha with a robust decrease in ribosome association with mRNA. The activity against T. brucei also implicates eIF2 alpha phosphorylation, as replacement of WT-eIF2 alpha with a non-phosphorylatable eIF2 alpha, or knocking down eIF2 protein kinase-3 by RNAi increased resistance to I-17. Therefore, we demonstrate that eIF2 alpha phosphorylation can be engaged to develop trypanosome-static agents in general, and particularly by interfering with activity of eIF2 kinases.

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Common bean (Phaseolus vulgaris L.) is one of the most important legumes for human consumption and is a staple food in the diet of the population of some countries of Latin America, Africa and Asia. The distinction between cultivars is based predominantly on morphological descriptors, which proved inefficient for the differentiation of some cultivars. This study had the objective of describing, distinguishing and evaluating the agronomic potential of 39 common bean cultivars of the carioca and black grain groups registered for cultivation in Brazil, based on 49 morphoagronomic descriptors and microsatellite (simple sequence repeat -SSR) markers. The morphoagronomic traits of each commercial group were characterized in four environments. Thirty-seven SSR markers were used for the molecular description. The morphological data, analyzed by the Shanonon-Weaver index, detected low variability among cultivars for qualitative data. On the other hand, the estimates of variance analysis, relative importance of the traits and hierarchical grouping analysis applied to the quantitative variables showed that the descriptors related to plant morphology were the most important for the carioca group, and those related to seed morphology were determining for the black group. The genetic parameters estimated for SSR markers by hierarchical and Bayesian cluster analysis identified 116 alleles, with 33 and 30 polymorphic loci and 24 and 22 private alleles for the carioca and black groups, respectively. The combined use of morphoagronomic and molecular descriptors improves the distinguishability of cultivars, contributing in a more efficient way to breeding and to the protection of cultivars.

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ABSTRACT Forty isolates of endophytic bacteria isolated from banana tree roots were assessed as to their capacity to solubilize phosphate in a solid culture medium supplemented with different inorganic and one organic source of phosphorus. The amount of phosphorus (P) in each liquid medium was quantified, and an indirect assessment of acid phosphatase activity was performed. All assays had a fully randomized design, with three repetitions. Approximately 67.5% of the 40 isolates assessed in solid medium solubilized phosphorus from tricalcium phosphate and 7.5% of the isolates solubilized phosphorus from soy lecithin; no isolates exhibited P solubilization capacity in medium supplemented with iron phosphate. Acid phosphatase activity was detected in 65% of the isolates; Aneurinibacillus sp. and Lysinibacillus sp. isolates presented with the best solubilization indexes. All of the assessed isolates exhibited a capacity to reduce the potential of hydrogen in liquid medium supplemented with tricalcium phosphate. Isolate EB. 78 (Bacillus sp.) exhibited P solubilization capacity in solid media when Ca3(PO4)2 and soy lecithin were used as P sources; this isolate significantly reduced the pH of the liquid medium and exhibited acid phosphatase activity. The results of the present study highlight isolates that exhibit variations in their capacity to solubilize P. These isolates should be used in future tests to assess their field performance.

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SUMMARY The study aimed to evaluate the effect of feed restriction on the development of organs and fat deposition in castrated Caninde goats. Twenty one goats were distributed in a completely randomized design, with initial body weight of 15.9 ± 1.03kg. The goats were allocated into three levels of feed restriction (seven animals per level): ad libitum; moderate restriction (restriction of 20% on the amount of feed consumed by the animals fed ad libitum) and severe restriction (40% restriction on the amount of feed consumed by the animals fed ad libitum). The diet consisted of 55% forage (Tifton) and 45% concentrate. At 110 days of experiment, the goats were slaughtered with abody weight of 23.5kg ± 2.5kg. The components included the internal organs (spleen, heart, liver, pancreas, lungs, kidneys, blood) the emptied and cleaned digestive tract and fat deposits (in heart, mesenteric, omental and pelvic-renal) were weighed. Feed restriction in Canindé goats affected the weight of organs and fat deposition in regions more involved with the energy storage function (mesenteric, omental and pelvic-renal) (P 0.05), however, feed restriction does not affect the percentage of the organs in relation to EBW (P > 0.05), indicating that even under feed restriction organ development is proportional to the development of the body.

RESUMO Objetivou-se avaliar o efeito da restrição alimentar sobre o desenvolvimento dos órgãos e deposição de gordura em caprinos Canindé castrados. Foram utilizados 21 cabritos confinados, em um delineamento inteiramente casualizado, com peso inicial de 15,9 ± 1,03kg. Os cabritos foram alocados em três níveis de restrição alimentar (sete animais por nível): ad libitum (alimentados à vontade); restrição moderada (restrição de 20% em relação à quantidade de matéria natural consumida pelos animais alimentados ad libitum) e restrição severa (restrição de 40% em relação à quantidade de ração consumida pelos animais alimentados ad libitum). A ração experimental apresentou uma proporção de 55% de volumoso (Tifton) e 45% de concentrado. Aos 110 dias de experimento os cabritos foram abatidos com peso médio de 23,5 kg ± 2,5 kg. Esvaziou-se o trato gastrointestinal (TGI), a bexiga e a vesícula biliar e foram mensurados os seus pesos para determinação do peso de corpo vazio (PCV). Foram separados e registrados os pesos dos órgãos (baço, coração, fígado, pâncreas, pulmões, rins, sangue, TGI) e dos depósitos de gordura (cardíaca, mesentérica, omental e pélvico-renal). A restrição alimentar em caprinos Canindé afetou o peso absoluto dos órgãos e das gorduras (P 0.05) que estão mais envolvidas com a função de reserva energética (mesentérica, omental e pélvico-renal), no entanto, não afetou o percentual dos órgãos em relação ao PCV (P > 0.05), indicando que mesmo sob restrição o desenvolvimento dos órgãos é proporcional ao desenvolvimento do corpo.

RESUMO

SUMMARY The study aimed to evaluate the effect of feed restriction on the development of organs and fat deposition in castrated Caninde goats. Twenty one goats were distributed in a completely randomized design, with initial body weight of 15.9 ± 1.03kg. The goats were allocated into three levels of feed restriction (seven animals per level): ad libitum; moderate restriction (restriction of 20% on the amount of feed consumed by the animals fed ad libitum) and severe restriction (40% restriction on the amount of feed consumed by the animals fed ad libitum). The diet consisted of 55% forage (Tifton) and 45% concentrate. At 110 days of experiment, the goats were slaughtered with abody weight of 23.5kg ± 2.5kg. The components included the internal organs (spleen, heart, liver, pancreas, lungs, kidneys, blood) the emptied and cleaned digestive tract and fat deposits (in heart, mesenteric, omental and pelvic-renal) were weighed. Feed restriction in Canindé goats affected the weight of organs and fat deposition in regions more involved with the energy storage function (mesenteric, omental and pelvic-renal) (P 0.05), however, feed restriction does not affect the percentage of the organs in relation to EBW (P > 0.05), indicating that even under feed restriction organ development is proportional to the development of the body.

RESUMO Objetivou-se avaliar o efeito da restrição alimentar sobre o desenvolvimento dos órgãos e deposição de gordura em caprinos Canindé castrados. Foram utilizados 21 cabritos confinados, em um delineamento inteiramente casualizado, com peso inicial de 15,9 ± 1,03kg. Os cabritos foram alocados em três níveis de restrição alimentar (sete animais por nível): ad libitum (alimentados à vontade); restrição moderada (restrição de 20% em relação à quantidade de matéria natural consumida pelos animais alimentados ad libitum) e restrição severa (restrição de 40% em relação à quantidade de ração consumida pelos animais alimentados ad libitum). A ração experimental apresentou uma proporção de 55% de volumoso (Tifton) e 45% de concentrado. Aos 110 dias de experimento os cabritos foram abatidos com peso médio de 23,5 kg ± 2,5 kg. Esvaziou-se o trato gastrointestinal (TGI), a bexiga e a vesícula biliar e foram mensurados os seus pesos para determinação do peso de corpo vazio (PCV). Foram separados e registrados os pesos dos órgãos (baço, coração, fígado, pâncreas, pulmões, rins, sangue, TGI) e dos depósitos de gordura (cardíaca, mesentérica, omental e pélvico-renal). A restrição alimentar em caprinos Canindé afetou o peso absoluto dos órgãos e das gorduras (P 0.05) que estão mais envolvidas com a função de reserva energética (mesentérica, omental e pélvico-renal), no entanto, não afetou o percentual dos órgãos em relação ao PCV (P > 0.05), indicando que mesmo sob restrição o desenvolvimento dos órgãos é proporcional ao desenvolvimento do corpo.

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Fixation of an osteochondral fracture after acute patellar dislocation is an infrequent form of treatment. Likewise, the location of this fragment in the lateral region of the lateral femoral condyle, functioning as a free body, is uncommon. The aim of this study was to present a case of osteochondral fracture of the patella at an unusual site, along with the therapy used and the clinical follow-up.

A fixação da fratura osteocondral após a luxação aguda da patela é um tratamento infrequente, bem como a localização desse fragmento na região lateral do côndilo femoral lateral que funciona como um corpo livre. O objetivo desta pesquisa foi apresentar um caso de fratura osteocondral da patela em sítio não usual, assim como a terapêutica adotada e o seguimento clínico.

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OBJECTIVES: to review radiographs of patients who suffered tendon tears of the knee extensor apparatus and observe alterations that might be factors predisposing toward this type of injury. METHODS: we retrospectively analyzed 60 cases of injury to the knee extensor mechanism that were treated surgically at the Miguel Couto Municipal Hospital between March 2004 and March 2011. Four patients were excluded due to poor quality of the examination. RESULTS: of the 56 patients evaluated, 23 were considered to be normal and 33 presented radiographic alterations. Among these, eight (24.3%) presented suprapatellar osteophytes alone; seven (21.2%), infrapatellar calcification; seven (21.2%), suprapatellar calcification; six (18.2%), supra- and infrapatellar osteophytes; and five (15.1%), infrapatellar osteophytes alone. CONCLUSION: radiographic alterations were frequently observed in patients with extensor mechanism tears.

OBJETIVOS: revisar radiografias de pacientes vítimas de rupturas tendinosas do aparelho extensor do joelho e observar alterações que possam ser um fator predisponente a esse tipo de lesão. MÉTODOS: analisamos, retrospectivamente, 60 lesões do mecanismo extensor do joelho, tratadas cirurgicamente no Hospital Municipal Miguel Couto, de março de 2004 a março de 2011, e quatro pacientes foram excluídos por má qualidade do exame. RESULTADOS: dos 56 pacientes avaliados, 23 foram considerados normais e 33 apresentaram alterações radiográficas, oito (24,3%) com presença de osteófito suprapatelar isolado, sete (21,2%) com calcificação infrapatelar, sete (21,2%) com calcificação suprapatelar, seis (18,2%) com osteófitos supra e infrapatelares e cinco (15,1%) com osteófito infrapatelar isolado. CONCLUSÃO: as alterações radiográficas foram frequentes em pacientes com ruptura do mecanismo extensor.

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The present study was conducted to evaluate the effect of 2-phenylethynyl-butyltellurium (PEBT), an organotellurium compound, at doses of 5 and 10 mg/kg on memory, employing the step-down inhibitory avoidance task in mice. Moreover, the involvement of glutamate uptake and release in cerebral cortex and hippocampus of mice was investigated. A single oral administration (p.o.) of PEBT at the dose of 10 mg/kg 1h before training (acquisition), immediately after training (consolidation) or 1 h before the test session (retrieval) of the step-down inhibitory avoidance task increased the step-through latency time in comparison to the control mice. In the open-field test, no significant differences in the number of crossings and rearings were observed among groups. The [(3)H]glutamate uptake by cerebral cortex and hippocampal slices of mice was significantly inhibited after 1h of treatment with PEBT. After 24h of PEBT exposure, only the hippocampal [(3)H]glutamate uptake was inhibited. The [(3)H]glutamate release by cerebral cortex and hippocampal synaptosomes of mice was not altered. These results suggest that PEBT improved memory stages (acquisition, consolidation and retrieval) in the step-down inhibitory avoidance task in mice. The improvement of memory by PEBT seems most likely to be mediated through an interaction with the amino acid transporters of the glutamatergic system.

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