Carbohydrate metabolism of Xylella fastidiosa: Detection of glycolytic and pentose phosphate pathway enzymes and cloning and expression of the enolase gene

The objective of this work was to assess the functionality of the glycolytic pathways in the bacterium Xylella fastidiosa. To this effect, the enzymes phosphoglucose isomerase, aldolase, glyceraldehyde-3-phosphate dehydrogenase and pyruvate kinase of the glycolytic pathway, and glucose 6-phosphate dehydrogenase of the Entner-Doudoroff pathway were studied, followed by cloning and expression studies of the enolase gene and determination of its activity. These studies showed that X. fastidiosa does not use the glycolytic pathway to metabolize carbohydrates, which explains the increased duplication time of this phytopatogen. Recombinant enolase was expressed as inclusion bodies and solubilized with urea (most efficient extractor), Triton X-100, and TCA. Enolase extracted from X. fastidiosa and from chicken muscle and liver is irreversibly inactivated by urea. The purification of enolase was partial and resulted in a low yield. No enzymatic activity was detected for either recombinant and native enolases, aldolase, and glyceraldehyde-3-phosphate dehydrogenase, suggesting that X. fastidiosa uses the Entner-Doudoroff pathway to produce pyruvate. Evidence is presented supporting the idea that the regulation of genes and the presence of isoforms with regulation patterns might make it difficult to understand the metabolism of carbohydrates in X. fastidiosa.

Effect of fluoride intake on carbohydrate metabolism, glucose tolerance, and insulin signaling

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

HETEROGENEOUS GROWTH IN THE NILE TILAPIA - SOCIAL STRESS AND CARBOHYDRATE-METABOLISM

Increase in heterogeneous growth as a result of grouping in the Nile tilapia, Oreochromis niloticus (L.), is presumed to be partially promoted by the social stress imposed by the dominant fish on the subordinates. Such stress may decrease the energy available for growth. In this study, the effect of social stress on carbohydrate metabolism was studied in adult, growing males. All animals were deprived of food during the experimentation period and pairing was imposed for either 2 or 4 days. Glycemia was measured before and after pairing, and muscle and liver glycogen contents were determined only after pairing. Subordinate fish showed the highest consumption of carbohydrate reserves. This response was caused by the social stress imposed which corroborates the idea that metabolic differences promoted by social stress may be involved in the rouping effect on heterogeneous growth in the Nile tilapia.

Effect of fasting on carbohydrate metabolism in frugivorous bats (Artibeus lituratus and Artibeus jamaicensis)

The compensatory changes of carbohydrate metabolism induced by fasting were investigated in frugivorous bats, Artibeus lituratus and Artibeus jamaicensis. For this purpose, plasma levels of glucose and lactate, liver and muscle glycogen content, rates of liver gluconeogenesis and the activity of related enzymes were determined in male bats. After a decrease during the first 48 h of fasting, plasma glucose levels remained constant until the end of the experimental period. Plasma lactate levels, extremely high in fed bats, decreased after 48 h of fasting. Similarly, liver glycogen content, markedly high in fed animals, was reduced to low levels after 24 h without food. Muscle glycogen was also reduced in fasted bats. The expected increase in liver gluconeogenesis during fasting was observed after 48 h of fasting. The activities of liver glucose-6-phosphatase and fructose-1,6-bisphosphatase were not affected by food withdrawn. on the other hand, fasting for 24 h induced an increase in the activity of liver cytosolic phosphoenolpyruvate carboxykinase. The data indicate that liver gluconeogenesis has an important role in the glucose homeostasis in frugivorous bats during prolonged periods of food deprivation. During short periods of fasting liver glycogenolysis seems to be the main responsible for the maintenance of glycemia. (C) 2005 Elsevier B.V. All rights reserved.

Growth performance and energetic metabolism of pacu, Piaractus mesopotamicus (Holmberg, 1887), fed diets supplemented with ammonium metavanadate

Vanadium compounds mimic most of the metabolic effects of insulin, suggesting that it might be useful to improve utilization of dietary carbohydrate. This work evaluated the effect of dietary ammonium metavanadate (H(4)NO(3)V) on the growth performance and energy metabolism of pacu, an omnivorous South America characin. Two hundred and eighty-eight fish were distributed into four blocks according to the body weight (21.8 +/- 1.7, 28.5 +/- 2.0, 28.4 +/- 1.9, 35.7 +/- 1.9 g), stocked in 24 plastic tanks and fed twice daily with isonitrogenous and isoenergetic diets containing six levels of H(4)NO(3)V (0, 10, 50, 100, 300 and 1000 mg kg(-1)) for 60 days. Increasing levels of dietary ammonium metavanadate did not improve growth (P > 0.05), and the highest level of inclusion (1000 mg kg(-1)) reduced performance (P < 0.05). Blood glucose levels decreased (P < 0.05) in fish fed 300 and 1000 mg kg(-1) H(4)NO(3)V, but no differences were observed in other blood metabolites. A slight increase in muscle lipid content was observed in fish fed a diet containing 300 mg kg(-1) H(4)NO(3)V. Based on the results of this study, there is no benefit in supplementing pacu diets with metavanadate.

Effects of physical training with different intensities of effort on lipid metabolism in rats submitted to the neonatal application of alloxan

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Energy Metabolism in Cardiac Remodeling and Heart Failure

Fatty acids are the main substrates used by mitochondria to provide myocardial energy under normal conditions. During heart remodeling, however, the fuel preference switches to glucose. In the earlier stages of cardiac remodeling, changes in energy metabolism are considered crucial to protect the heart from irreversible damage. Furthermore, low fatty acid oxidation and the stimulus for glycolytic pathway lead to lipotoxicity, acidosis, and low adenosine triphosphate production. While myocardial function is directly associated with energy metabolism, the metabolic pathways could be potential targets for therapy in heart failure. © 2013 by Lippincott Williams & Wilkins.

Progresso científico sobre nutrição de animais de companhia na primeira década do século XXI

O conhecimento científico sobre nutrição de animais de companhia tem aumentado de forma contínua, acompanhando o fenômeno visto em diversas áreas de conhecimento. Na última década, têm-se prioritariamente pesquisas direcionadas ao uso de nutrientes na promoção de saúde, prevenção de doenças degenerativas, melhoria da qualidade de vida e aumento da expectativa de vida de cães e gatos. Este direcionamento de pesquisas é, em grande parte, explicado pela importância que cães e gatos assumiram na vida das pessoas, fazendo com que as decisões alimentares dos proprietários com seus animais se assemelhassem às que adotam para si próprios. A publicação da nova revisão do Nutrient Requirements of Dogs and Cats em 2006 foi, talvez, o avanço recente mais significativo, trazendo novo entendimento sobre necessidades energéticas e de nutrientes nas diferentes fases de desenvolvimento e estados fisiológicos. Apesar destes avanços, ainda são necessárias pesquisas na área de caracterização físico-química e de utilização dos ingredientes, efeitos do processo de extrusão e mesmo de necessidades nutricionais, que conta com uma base pequena de artigos disponíveis, a maioria antiga. Talvez os desafios científicos mais importantes em nutrição de cães e gatos sejam metabolismo de carboidratos, importância da massa corporal magra na saúde, urolitíases, gerontologia, relação entre microbiota intestinal e saúde, imunonutrição e manejo nutricional em condições clínicas específicas importantes para estas espécies.

Cissus sicyoides (princess vine) in the long-term treatment of streptozotocin-diabetic rats

Leaf decoctions of Cissus sicyoides (princess vine) are taken widely as a popular remedy for diabetes mellitus in Brazil, where its common name is 'vegetal insulin'. However, there have been practically no attempts so far to determine scientifically whether it has antidiabetic effects and we decided to administer leaf decoctions, over extended periods, to normal and streptozotocin-diabetic rats, and investigate the effects of this treatment on the physiological and metabolic parameters that are altered in diabetic animals. The experimental model adopted was shown to be appropriate by running a parallel treatment with insulin, which led to expected improvements in several abnormal parameter values. The decoction treatment significantly reduced the intake of both food and fluid and the volume of urine excreted, as well as the levels of blood glucose, urinary glucose and urinary urea, in comparison with controls. Lipid metabolism was not affected by the treatment; nor was the level of hepatic glycogen in diabetic animals, which indicated that the mechanism responsible for the improvement in carbohydrate metabolism, observed in animals treated with the decoction, could not involve inhibition of glycogenolysis and/or stimulation of glycogenesis. The fact that normal animals treated with C. sicyoides exhibited no changes in any of the measured parameters suggests that its mode of action in diabetic animals does not resemble those of sulphonylurea or insulin. It may, however, act in a similar way to biguanide, via inhibition of gluconeogenesis.

Anti-diabetic activity of Bauhinia forficata decoction in streptozotocin-diabetic rats

The effects of using Bauhinia forficata leaf decoction (150 g leaf/l water; 35.2+/-7.8 ml/100 g body weight mean daily dose) as a drinking-water substitute for about I month on streptozotocin-diabetes (STZ-diabetes) in male Wistar rats were investigated. The physico-metabolic parameters measured were: body weight, food and liquid intake, urinary volume, hepatic glycogen, serum triglycerides and cholesterol, plasma glucose, urinary glucose and urea, and the weight of epididymal and retroperitoneal adipose tissue and soleus and extensor digitorum longus muscles. The STZ-diabetic rats treated with decoction showed a significant reduction in serum and urinary glucose and urinary urea as compared to the STZ-diabetic control, no difference being seen between decoction-treated and -untreated non-diabetic rats. The other physico-metabolic factors showed no changes in treated STZ-diabetic rats. The improvement in carbohydrate metabolism seen in the rats treated with Bauhinia forficata decoction does not appear to be linked to the inhibition of glycogenolysis or the stimulation of glycogenesis nor does it appear to act in a way similar to insulin or the sulfonylureas, although it may act by the inhibition of neoglycogenesis in a manner similar to that of the biguanides. (C) 2002 Elsevier B.V. Ireland Ltd. All rights reserved.

Metabolic responses to glucoprivation induced by 2-deoxy-D-glucose in Brycon cephalus (Teleostei, Characidae)

The effects of functional cytoglucopenia provoked by 2-deoxy-D-glucose (2-DG) were studied in adult Brycon cephalus, an omnivorous fish from the Amazon Basin in Brazil. Glycogen content in liver and muscle as well as plasmatic glucose, free fatty acids (FFA), insulin, and glucagon were measured. After 48 h fasting, an intraperitoneal saline injection (NaCl 0.6 g/100 ml) was administered to control fish, whereas the experimental group received 2-DG, dissolved in saline, in the dosage of 80 mg/kg (0.487 mmol/kg) or 150 mg/kg (0.914 mmol/kg) body weight; injection volume was 5 ml in all treatments. Blood and tissue samples were taken immediately before, and 2, 8, 10, and 24 h after administration of the drug or saline. Fish injected with both doses of 2-DG showed a marked increase in glycemia levels. Liver and muscle glycogen decreased after 2-DG administration and reached their lowest values 10-24 h after injection, while in control animals no significant changes were observed. Elevation in plasma glucagon was observed only in response to the maximum dosage of 2-DG administered, especially 10 h and 24 h post-injection. Plasma insulin levels were lower in animals treated with the glucose analogue but only statistically significant 24 h after drug administration. In conclusion, the administration of the non-metabolizable glucose analogue 2-DG in B. cephalus is a stimulus to generate responses towards an increase in the glucose available to tissues, which is a characteristic of a fasting situation. All the above data support the interest of 2-DG administration as a model to study carbohydrate metabolism adjustment mechanisms in fish.

The use of the oxidative stress responses as biomarkers in Nile tilapia (Oreochromis niloticus) exposed to in vivo cadmium contamination

Water contaminants have a high potential risk for the health of populations. Protection from toxic effects of environmental water pollutants primarily involves considering the mechanism of low level toxicity and likely biological effects in organisms who live in these polluted waters. The biomarkers assessment of oxidative stress and metabolic alterations to cadmium exposure were evaluated in Nile tilapia, Oreochromis niloticus. The fish were exposed to 0.35, 0.75, 1.5, and 3.0 mg/l concentrations of Cd2+ (CdCl2) in water for 60 days. Fish that survived cadmium exposure showed a metabolic shift and a compensatory development for maintenance of the body weight gain. We observed a decreased glycogen content and decreased glucose uptake in white muscle. Lactate dehydrogenase (LDH) and creatine phosphokinase (CK) activities were also decreased, indicating that the glycolytic capacity was decreased in this tissue. No alterations were observed in total protein content in white muscle due to cadmium exposure suggesting a metabolic shift of carbohydrate metabolism to maintenance of the muscle protein reserve. There was an increase in glucose uptake, CK increased activity, and a clear increase of LDH activity in red muscle of fish with cadmium exposure. Since no alterations were observed in lipoperoxide concentration, while antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were changed in the liver and the red and white muscle of fish with cadmium exposure, we can conclude that oxygen free radicals are produced as a mediator of cadmium toxicity. Resistance development is related with increased activities of antioxidant enzymes, which were important in the protection against cadmium damage, inhibiting lipoperoxide formation. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Low-Resolution Molecular Models Reveal the Oligomeric State of the PPAR and the Conformational Organization of Its Domains in Solution

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)