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ABSTRACTThe brain aging process triggers cognitive function impairment, such as memory loss and compromised quality of life. Cognitive impairment is based on bioenergetic status, with reduced glucose uptake and metabolism in aged brains. Anaplerotic substrates are reported to promote mitochondrial ATP generation, having been tested in clinical trials for the treatment of neurological disorders and metabolic diseases.Objectives and Methods: To assess whether the improvement in oxidative capacity ameliorates cognitive function in adults (12 weeks), and aged (22-month-old) C57/6BJ mice, they received (1) a ketogenic diet, (2) a ketogenic diet supplemented with the anaplerotic substance, triheptanoin, or (3) a control diet for 12 weeks. Spontaneous alternation and time spent in a previously closed arm in the Y-maze test and time interacting with an unknown object in the novel object recognition test (NORT) were used to evaluate working memory. Acetylcholinesterase (AChE) activity in the prefrontal lobe, brain left hemisphere, and cerebellum was also evaluated. Glucose transporter 3 (GLUT3) expression in the prefrontal lobe was analyzed by western blotting.Results: The ketogenic diet (KD) reduced spontaneous alternation in aged mice, leading to lower AChE activity in the aged prefrontal lobe and cerebellum, and in the parieto-temporal-occipital lobe of adult mice. Furthermore, KD decreased GLUT3 protein expression in the frontal lobe of the adults.Discussion: Supplementation of KD with triheptanoin prevented memory impairment and showed similar values of AChE activity and GLUT3 expression compared to the controls. Our data suggest that triheptanoin has a potential role in the bioenergetic capacity of the brain, improving cognitive function.
Assuntos
Acetilcolinesterase , Qualidade de Vida , Camundongos , Animais , Transportador de Glucose Tipo 3/metabolismo , Acetilcolinesterase/metabolismo , Triglicerídeos , Encéfalo/metabolismo , CogniçãoRESUMO
Retinocollicular connections form precise topographical maps that are normally completed through the selective elimination of misplaced axons and the stabilization of topographically ordered axon terminals during early development. Omega-3 fatty acids, acquired exclusively through the diet, and its main metabolite, docosahexaenoic acid (DHA), are involved in brain development and synaptic maturation. We have previously shown that the nutritional restriction of omega-3/DHA results in abnormal retinocollicular topographical fine-tuning. Therefore, we studied the role of omega-3 fatty acids nutritional supplementation and the developmental time windows during which this postnatal supplementation would restore normal topographical maps in the visual system. Female rats and their litters were chronically fed with either control (soy oil) or restricted omega-3 (coconut oil) diets. Fish oil supplementation was introduced between either postnatal day (PND) 7-13, PND7-28 or PND21-42. At PND13, PND28 or PND42, animals received an anterograde eye injection of a neuronal tracer to visualize retinocollicular axons. Confirming previous observations we found that an omega-3/DHA deficiency resulted in an abnormally high innervation density of retinal axons at the visual layers of the superior colliculus (SC). Although a short-term fish oil supplementation between PND7-13 could not restore normal retinocollicular topography, an extended treatment between PND7-28 completely recovered normal innervation densities of retinotectal axons. However, a late onset supplementation protocol, between PND28-42, was no longer effective in the restoration of the abnormal topographical pattern induced by an early omega-3 nutritional malnutrition. The results suggest a critical period for omega3/DHA dietary intake for the proper development of visual topographical maps.
Assuntos
Ácidos Graxos Ômega-3/administração & dosagem , Retina/crescimento & desenvolvimento , Colículos Superiores/crescimento & desenvolvimento , Vias Visuais/crescimento & desenvolvimento , Animais , Suplementos Nutricionais , Ácidos Docosa-Hexaenoicos/administração & dosagem , Feminino , Ratos , Retina/citologia , Colículos Superiores/citologia , Fatores de TempoRESUMO
An ATP diphosphohydrolase (EC 3.6.1.5) activity was identified in a Leishmania (Viannia) braziliensis promastigotes preparation (Lb). Ultrastructural cytochemical microscopy showed this protein on the parasite surface and also stained a possible similar protein at the mitochondrial membrane. Isolation of an active ATP diphosphohydrolase isoform from Lb was obtained by cross-immunoreactivity with polyclonal anti-potato apyrase antibodies. These antibodies, immobilized on Protein A-Sepharose, immunoprecipitated a polypeptide of approximately 48 kDa and, in lower amount, a polypeptide of approximately 43 kDa, and depleted 83% ATPase and 87% of the ADPase activities from detergent-homogenized Lb. Potato apyrase was recognized in Western blots by IgG antibody from American cutaneous leishmaniasis (ACL) patients, suggesting that the parasite and vegetable proteins share antigenic conserved epitopes. Significant IgG seropositivity in serum samples diluted 1:50 from ACL patients (n=20) for Lb (65%) and potato apyrase (90%) was observed by ELISA technique. Significant IgG antibody reactivity was also observed against synthetic peptides belonging to a conserved domain from L. braziliensis NDPase (80% seropositivity) and its potato apyrase counterpart (50% seropositivity), in accordance with the existence of shared antigenic epitopes and demonstrating that in leishmaniasis infection the domain r82-103 from L. braziliensis NDPase is a target for the human immune response.
Assuntos
Apirase/metabolismo , Leishmania braziliensis/enzimologia , Leishmaniose Cutânea/parasitologia , Sequência de Aminoácidos , Animais , Anticorpos Antiprotozoários/sangue , Apirase/genética , Apirase/imunologia , Western Blotting , Humanos , Imunoprecipitação , Isoenzimas , Leishmania braziliensis/imunologia , Leishmania braziliensis/ultraestrutura , Leishmaniose Cutânea/imunologia , Microscopia Eletrônica , Dados de Sequência MolecularRESUMO
A Leishmania (Leishmania) amazonensis ATP diphosphohydrolase isoform was partially purified from plasma membrane of promastigotes by preparative non-denaturing polyacrylamide gel electrophoresis. SDS-PAGE followed by Western blots developed with polyclonal anti-potato apyrase antibodies identified diffuse bands of about 58-63 kDa, possibly glycosylated forms of this protein. By ELISA technique, a significantly higher total IgG antibody level against potato apyrase was found in serum from promastigote-infected mice, as compared to the uninfected mice, confirming both the existence of shared epitopes between the parasite and vegetable proteins, and the parasite ATP diphosphohydrolase antigenicity. By Western blotting, serum from amastigote-infected BALB/c mice recognizes both potato apyrase and this antigenic ATP diphosphohydrolase isoform isolated from promastigotes, suggesting that it is also expressed in the amastigote stage. The infection monitored along a 90-day period in amastigote-infected mice showed reactivity of IgG2a antibody in early steps of infection, while the disappearance of the IgG2a response and elevation of IgG1 antibody serum levels against that shared epitopes were associated with the progression of experimental leishmaniasis. This is the first observation of the antigenicity of a L. (L.) amazonensis ATP diphosphohydrolase isoform, and of the ability of cross-immunoreactivity with potato apyrase to differentiate serologically stages of leishmaniasis in infected mice.
Assuntos
Apirase/imunologia , Leishmania mexicana/enzimologia , Leishmaniose Cutânea/diagnóstico , Solanum tuberosum/enzimologia , Animais , Variação Antigênica , Apirase/isolamento & purificação , Apirase/metabolismo , Western Blotting , Reações Cruzadas , Progressão da Doença , Eletroforese em Gel de Poliacrilamida , Epitopos , Feminino , Isoenzimas/imunologia , Isoenzimas/isolamento & purificação , Isoenzimas/metabolismo , Camundongos , Camundongos Endogâmicos BALB CRESUMO
We investigated in young rats the effects of malnutrition on the main structures of the circadian timing system: retina, hypothalamic suprachiasmatic nuclei (SCN), thalamic intergeniculate leaflet, retinohypothalamic- and geniculohypothalamic tracts. Control rats were born from mothers fed a commercial diet since gestation, and malnourished rats from mothers fed a multideficient diet since gestation (GLA group) or lactation (LA group). After weaning, pups received the same diet as their mothers, and were analysed at postnatal days 27, 30-33 and 60-63. Brain sections were processed to visualise in the SCN neuropeptide Y immunoreactivity and terminal labeling after intraocular tracer injections. Nissl staining was used to assess cytoarchitectonic boundaries of the SCN and cell features in retinal whole mounts. Cell counts, morphometric and densitometric analysis were performed. Compared with controls, the total retinal surface was reduced and the topographical distribution of retinal ganglion cells was altered in malnourished rats, with changes in their density. Alterations were also detected in the SCN dimensions in the GLA and LA groups at one and two postnatal months, as well as in the SCN portion occupied by the retinal input in the GLA group at days 30-33, but not in the NPY-containing geniculohypothalamic tract. The present data point to subtle changes, with a low and differential vulnerability to early malnutrition, of structures involved in circadian timing regulation. Furthermore, the present findings suggest that the altered circadian rhythmicity previously documented in malnourished rats cannot be ascribed to impaired development of the retino- and geniculohypothalamic projections to the SCN.
Assuntos
Encéfalo/patologia , Corpos Geniculados/patologia , Desnutrição/patologia , Retina/patologia , Núcleo Supraquiasmático/patologia , Fatores Etários , Animais , Animais Recém-Nascidos , Peso Corporal/fisiologia , Encéfalo/citologia , Encéfalo/crescimento & desenvolvimento , Encéfalo/metabolismo , Contagem de Células/métodos , Tamanho Celular , Feminino , Corpos Geniculados/anatomia & histologia , Corpos Geniculados/crescimento & desenvolvimento , Corpos Geniculados/metabolismo , Imuno-Histoquímica/métodos , Lactação , Masculino , Vias Neurais/crescimento & desenvolvimento , Vias Neurais/patologia , Neurônios/metabolismo , Neurônios/patologia , Neuropeptídeo Y/metabolismo , Tamanho do Órgão/fisiologia , Ratos , Ratos Wistar , Retina/citologia , Retina/crescimento & desenvolvimento , Retina/metabolismo , Fatores Sexuais , Coloração e Rotulagem/métodos , Núcleo Supraquiasmático/crescimento & desenvolvimento , Núcleo Supraquiasmático/metabolismoRESUMO
As plantas denominadas de ruibarbo sintetizam antraquinonas e taninos, que são responsáveis pelos efeitos laxante e adstringente, respectivamente. Análises da qualidade de cinco matérias-primas de ruibarbo foram realizadas a fim de detectar adulterações. As reações para antraquinonas e taninos foram positivas. Os constituintes rapônticos foram observados na amostra de Rheum palmatum (2). As medidas de cinzas totais para Rheum palmatum (2) e Ferraria cathartica estão acima do esperado. Conforme resultados, as amostras de Rheum palmatum (2) e Ferraria cathartica devem estar adulteradas.
Plants known as rhubarb synthesize anthraquinones and tannins. Theses compounds have medicinal uses as laxative and astringent, respectivity. The quality of five raw materials was analysed to detect adulterant. Anthraquinones and tannins reactions were positive. Derivatives rhapontocoside were observed in the sample of Rheum palmatum (2). Total ashes were increased to Rheum palmatum (2) and Ferraria cathartica. According to the results, samples of Rheum palmatum (2) and Ferraria cathartica shoulde are adulterated.
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Vanillosmopsis erythropappa é um vegetal rico em óleos essenciais, especialmente o á-bisabolol. O objetivo deste estudo foi pesquisar a variação sazonal qualitativa de substâncias naturais, teores de óleos essenciais, cinzas totais, umidade e pH. Os resultados mostraram uma variação de flavonóides, taninos, triterpenóides, esteróides, saponinas, óleos essenciais, cinzas totais, umidade e pH, demonstrando que os aspectos ambientais influenciam o metabolismo dessa espécie. As médias anuais dos valores obtidos foram: óleos essenciais= 0,29 ± 0,09 por cento; cinzas totais= 3,13 ± 0,49 por cento; umidade= 6,56 ± 0,80 por cento e pH= 5,40 ± 0,24.
Vanillosmopsis erythropappa is a plant rich in essential oils, especially á-bisabolol. The aim of this study was research the qualitative seasonal variation of natural substances, tenor of essential oils, total ashes, dampness and pH. The results showed a variation of the flavonoids, tannins, triterpenoids, steroids, saponins, essential oils, total ashes, dampness and pH, demonstrating that environmental aspects influence on the metabolism of this specie. The annual averages of the obtained values were: essential oils = 0,29 ± 0,09 percent; total ashes = 3,13 ± 0,49 percent; dampness= 6,56 ± 0,80 percent and pH = 5,40 ± 0,24.
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The distribution of ganglion cell densities and sizes was studied in Nissl-stained flat-mount retinae fo the two-toed sloth. The area centrailis, a weak specializtion with low ganglion cell density, is located in the temporal retina close to the center of the eye. The presence of a visual treak was nsoted. The distribution of different ganglion cell sizes was approximately equal throughout the retina. Although the retinal organization differs from that of the closely related three-toed sloth, presumed function of retinl specializations in both species is to guide limb movements by permiting visualization of the branch along which the animal is climbing
Assuntos
Animais , Gânglios Simpáticos/citologia , Retina/citologia , Bichos-Preguiça/anatomia & histologia , Xenarthra/anatomia & histologia , Adaptação Fisiológica , Retina/fisiologiaRESUMO
1. The distribution ans size of retinal ganglion cells labelled with horseradish preoxidase (HRP) were studied in flat-mounted retinas of three-toed sloths. 2. Massive injections of HRP solution were made throughout the thalamus and midbrain in anesthetized sloths in order to retrogradely label the retinal ganglion cell population. Twenty to thirty h later the eyes were and the retinas flat-mounted and reacted with phenylenediamine-HCL and H2O2 to label ganglion cells,thus distinguishing then from other cells int he same retinal layer. 3. Ganglion cell density graually increased from about 500 cell/mm2 at the far periphery to a peak of about 1,500 cells/mm2 in an area, termed the area centralis, deep in the inferior temporal retina. The presence of a vertical visual steak was also noted. 4. The area centralis contained a higher frequency of small ganglion cells than the peripheral retina where large cells preominated. 5. /the unusual postion of the area centralis and cisual streak in the retina can be explained by the slot's unique ability to rotate its head 180- while climbing upside-dow along horizontal branches so that the head is right-side-up. If it is assumed that the branch directly above the sloth's head needs to be visualized for accurate claw placement then the branch would be imaged on the inferior temporal retina in an area corresponding to the maximum density region