RESUMO
Taguchi method was used to optimize loop mediated isothermal amplification tests aimed to amplify segments of the elongation factor 1a1 (tf-ef1a1), the 5,8 ribosomal gene (tf-5,8 r) or the beta tubulin 2 (tf-btub2) from the protozoan parasite Tritrichomonas foetus. L9 orthogonal array and quadratic loss functions that penalize deviations from prediction values revealed the effect of amplification reaction components. Analysis of variance (ANOVA) decomposed the contribution of individual factors to a small Ct. Confirmation experiments established that optimum conditions were predictable, verifiable and reproducible. Primers concentration conditioned the non-specific amplification of tf-ef1a1 while betaine and magnesium concentration contributed to accelerate the time to reach a positive threshold in tf-ef1a1, tf-5,8 r and tf-btub2. The general strategy of simple and robust experimental design holds potential as a general optimization protocol for LAMP tests in every diagnostic laboratory.
Assuntos
Técnicas de Diagnóstico Molecular , Técnicas de Amplificação de Ácido Nucleico , Tritrichomonas foetus , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/parasitologia , Primers do DNA , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Infecções Protozoárias em Animais/diagnóstico , Infecções Protozoárias em Animais/parasitologia , Sensibilidade e Especificidade , Tritrichomonas foetus/genéticaRESUMO
In this work, we report a characterization of gene expression profiles of mice hippocampus by use microarray after treatment with Psidium cattleyanum fruit (1000 mg/kg oral). After eight months, no toxic effects were detected with the supplementation. Genes differentially expressed include genes involved in a wide range of physiological functions, such as metabolism, transport, signal transduction and a group of genes whose function has not yet been identified. These results suggest that global analysis of gene expression might be useful to elucidate the mechanisms of beneficial phytochemical action and may also help to identify potential targets for further investigation.
El objetivo de éste trabajo fue evaluar el efecto de la administración oral prolongada (1000 mg/kg, 8 meses) del extracto de Psidium cattleyanum Sabine (Myrtaceae) en el perfil de la expresión génica en hipocampo de ratones idosos (Swiss). Después de 8 meses de suplementacion, no se detectaron efectos tóxicos en los animales tratados con relación al grupo control. Los genes con expresión diferencial incluyen, genes que codifican proteínas relacionadas con procesos de señalización, transcripción, metabolismo, así como genes con función desconocida. Los resultados demuestran la importancia de los microarray como herramienta para el estudio del mecanismo de acción de los compuestos fitoquímicos.
Assuntos
Masculino , Animais , Camundongos , Envelhecimento , Expressão Gênica , Extratos Vegetais/administração & dosagem , Hipocampo , Psidium/química , Análise em Microsséries , Brasil , PolifenóisRESUMO
The aim of this study was to carry out pharmacological screening in order to evaluate the potential effects of lyophilized fruits of different cultivars of Vaccinium ashei Reade (Family Ericaceae) berries, commonly known as rabbiteye blueberries, on nociception. This was achieved using the formalin, hot plate, tail-flick, and writhing tests in mice. During this experiment the mice consumed approximately 3.2-6.4 mg/kg/day (p.o.) of the anthocyanins. The extract was administered for 21 days or 60 minutes before test. Morphine and diclofenac (10 mg/kg, p.o.) as the standard drug (positive control) and water (via oral gavage) as the negative control were administered before all tests. The blueberry extract produced a significant decrease in constrictions induced by acetic acid and caused graded inhibition of the second phase of formalin-induced pain. Moreover, in both the hot plate and tail-flick tests, it significantly increased the threshold. These data suggest that the extract from V. ashei produced antinociceptive effects, as demonstrated in the experimental models of nociception in mice. Additional experiments are necessary in order to clarify the true target for the antinociceptive effects of rabbiteye blueberry extract.
Assuntos
Analgésicos/uso terapêutico , Antocianinas/uso terapêutico , Mirtilos Azuis (Planta)/química , Dor/tratamento farmacológico , Fitoterapia , Extratos Vegetais/uso terapêutico , Ácido Acético , Analgésicos/farmacologia , Animais , Antocianinas/farmacologia , Diclofenaco/farmacologia , Diclofenaco/uso terapêutico , Modelos Animais de Doenças , Formaldeído , Frutas , Temperatura Alta , Masculino , Camundongos , Morfina/farmacologia , Morfina/uso terapêutico , Contração Muscular , Dor/etiologia , Limiar da Dor/efeitos dos fármacos , Extratos Vegetais/farmacologiaRESUMO
Thyroid hormones (THs) have a relevant action on brain development and maintenance. By using an acute treatment to induce a hyperthyroid animal model, we aimed at investigating the effect of an altered THs levels on learning and memory and on the activity of Na(+), K(+)-ATPase in the rat brain. Our results have shown that the acute treatment with L-T4 did not alter the retrieval of the inhibitory avoidance task, but had a significant effect on the elevated plus maze and on open-field performance in rats. We suggest that animals subjected to L-T4 administration improved the habituation to a novel environment as well as a better evaluation of a dangerous environment, respectively. Na(+), K(+)-ATPase activity is increased in parietal cortex (30%), but it is not altered in hippocampus in L-T4 treated group. These both brain structures are involved in memory processing and it was previously demonstrated that there is a double dissociation between them for spatial location information, perceptual and episodic memory. We propose the hypothesis that this increase of Na(+), K(+)-ATPase activity in parietal cortex may be correlated to our results in behavior tests, which suggest a role of THs as well as of the Na(+), K(+)-ATPase in the cognitive process.
Assuntos
Comportamento Animal/efeitos dos fármacos , Encéfalo/enzimologia , Habituação Psicofisiológica/efeitos dos fármacos , Memória/efeitos dos fármacos , Assunção de Riscos , ATPase Trocadora de Sódio-Potássio/metabolismo , Tiroxina/farmacologia , Animais , Ansiedade/psicologia , Aprendizagem da Esquiva/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Hipocampo/enzimologia , Masculino , Atividade Motora/efeitos dos fármacos , Lobo Parietal/efeitos dos fármacos , Lobo Parietal/enzimologia , Ratos , Ratos Wistar , Membranas Sinápticas/efeitos dos fármacos , Membranas Sinápticas/enzimologia , Hormônios Tireóideos/sangue , Tiroxina/sangueRESUMO
Studies have shown that supplementation with berries rich in anthocyanins are effective in reducing oxidative stress associated with aging, and are beneficial in reversing age-related neuronal and behavioral changes. However, there are few reports on other biological activities of these polyphenols, such as genoprotective effects. The present experiments were performed to study the possible effects of 30-day administration of a lyophilized extract of Vaccinium ashei berries on cognitive performance using step-down inhibitory avoidance, open-field habituation and elevated plus-maze tasks, as well as on DNA damage in the hippocampus and cerebral cortex. The present study showed that the extract significantly enhanced long-term memory in the inhibitory avoidance task, induced an increase in the number of crossings during open-field habituation and had an anxiolytic effect in the elevated plus-maze task. Moreover, the extract reduced oxidative DNA damage in brain tissue in vitro. These results suggest that supplementation with V. ashei berries to mice improves performance on memory tasks and has a protective effect on DNA damage, possibly due to the antioxidant activity of polyphenols, including anthocyanins.
Assuntos
Antocianinas/farmacologia , Comportamento Animal/efeitos dos fármacos , Mirtilos Azuis (Planta) , Dano ao DNA/efeitos dos fármacos , Frutas , Extratos Vegetais/farmacologia , Animais , Ansiolíticos/farmacologia , Aprendizagem da Esquiva/efeitos dos fármacos , Córtex Cerebral/efeitos dos fármacos , Ensaio Cometa , Habituação Psicofisiológica/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , CamundongosRESUMO
Epidemiological studies suggest that diets with a high intake of vegetables and fruits may reduce the incidence of degenerative disorders including Alzheimer's disease. Berries are some of the popular fruits consumed worldwide. They are considered to be rich in anthocyanin pigments, a group belonging to the flavonoids, a widespread class of phenolic compounds. Anthocyanins have notorious pharmacological properties, and have been used in humans for therapeutic purposes. The present experiments were performed to study the possible effects of prolonged administration of lyophilised Vaccinium berries (blueberry, bilberry) on cognitive performance using step-down inhibitory avoidance, open field, elevated plus-maze, and radial maze tasks. During this experiment the rats consumed approximately 3.2 mg kg(-1)day (oral), of the anthocyanins. The lyophilised berries were administered for 30 days before first training. The present study showed that lyophilised berries significantly enhanced short-term memory, but not long-term memory in the inhibitory avoidance task, and induced an increase in the number of crossings in the first exposure to the open field. However, treated rats did not present any improvement of memory retention in open field habituation. Additionally, prolonged treatment with lyophilised berries did not have any significant effects in the elevated plus-maze task. Another interesting finding was that lyophilised berries improved working memory in the radial maze, with significant differences observed during sessions 1-2 and 4, but did not alter reference memory in this task. These results suggest that lyophilised berries may be beneficial in the prevention of memory deficits, one of the symptoms related to AD, and corroborate previous findings showing that flavonoids present effects in several learning paradigms.
Assuntos
Ansiedade/tratamento farmacológico , Mirtilos Azuis (Planta) , Memória/efeitos dos fármacos , Atividade Motora/efeitos dos fármacos , Fitoterapia , Vaccinium , Animais , Aprendizagem da Esquiva/efeitos dos fármacos , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Ratos , Ratos WistarRESUMO
Male Wistar rats were exposed to one-trial step-down inhibitory avoidance training using a 0.5 mA footshock. Through bilaterally implanted indwelling cannulae, they received bilateral 0.5 microL infusions of saline, mecamylamine (1.0 or 10.0 microg/side), or nicotine (0.6 or 3.0 microg/side) into the basolateral complex of the amygdaloid nucleus (BLA). Infusions were either 10 min before training (Experiment 1) or 4 min after training (Experiment 2). In Experiment 1, the animals were tested three times: first for working memory (WM) 5 s after training, then for short-term memory (STM) 90 min later, and finally for long-term memory (LTM) 24 h later. Mecamylamine depressed and nicotine enhanced WM, STM, and LTM. In Experiment 2, the treatments were given after WM was presumably over. Again, mecamylamine inhibited and nicotine enhanced STM and LTM. The results indicate that nAChRs in BLA participate in the regulation of WM formation and STM and LTM acquisition and consolidation.
Assuntos
Tonsila do Cerebelo/efeitos dos fármacos , Aprendizagem da Esquiva/efeitos dos fármacos , Mecamilamina/farmacologia , Memória de Curto Prazo/efeitos dos fármacos , Nicotina/farmacologia , Antagonistas Nicotínicos/farmacologia , Receptores Nicotínicos/efeitos dos fármacos , Retenção Psicológica/efeitos dos fármacos , Tonsila do Cerebelo/fisiologia , Animais , Aprendizagem da Esquiva/fisiologia , Relação Dose-Resposta a Droga , Esquema de Medicação , Potenciação de Longa Duração/efeitos dos fármacos , Potenciação de Longa Duração/fisiologia , Masculino , Memória de Curto Prazo/fisiologia , Ratos , Ratos Wistar , Receptores Colinérgicos/efeitos dos fármacos , Receptores Colinérgicos/fisiologia , Receptores Nicotínicos/fisiologia , Retenção Psicológica/fisiologiaRESUMO
Rats were bilaterally implanted with indwelling cannulae in the CA1 region of the dorsal hippocampus. After recovery from surgery, they were trained in a one-trial, step-down inhibitory avoidance task using a 0.5 mA foot shock. The animals received intrahippocampal infusions of either vehicle or anandamide (100 microM, 0.5 microl/side) 30 min before training. Then, either immediately post-training or 3 h later, they received infusions of saline, noradrenaline (0.5 microg/side), SKF 38393 (1.5 microg/side), oxotremorine (0.6 microg/side) or Sp-cAMPs (0.5 microg/side) also in the hippocampus. All animals were tested for retention 24-h post-training. Anandamide produced anterograde amnesia. Immediate, but not delayed, post-training treatment with Sp-cAMPs and noradrenaline reversed this effect. SKF 38393 and oxotremorine had no influence on the amnesia caused by anandamide either when given immediately or 3 h after training. The results suggest that the amnesic effect of anandamide is related to the known noradrenergic regulation of cAMP-dependent protein kinase (PKA) activity previously described in the hippocampus immediately after avoidance training, which is crucial to long-term memory (LTM) formation.