Skin fibroblast model to study an impaired glutathione synthesis: consequences of a genetic polymorphism on the proteome.

An impaired glutathione (GSH) synthesis was observed in several multifactorial diseases, including schizophrenia and myocardial infarction. Genetic studies revealed an association between schizophrenia and a GAG trinucleotide repeat (TNR) polymorphism in the catalytic subunit (GCLC) of the glutamate cysteine ligase (GCL). Disease-associated genotypes of this polymorphism correlated with a decrease in GCLC protein expression, GCL activity and GSH content. To clarify consequences of a decreased GCL activity at the proteome level, three schizophrenia patients and three controls have been selected based on the GCLC GAG TNR polymorphism. Fibroblast cultures were obtained by skin biopsy and were challenged with tert-butylhydroquinone (t-BHQ), a substance known to induce oxidative stress. Proteome changes were analyzed by two dimensional gel electrophoresis (2-DE) and results revealed 10 spots that were upregulated in patients following t-BHQ treatment, but not in controls. Nine corresponding proteins could be identified by MALDI mass spectrometry and these proteins are involved in various cellular functions, including energy metabolism, oxidative stress response, and cytoskeletal reorganization. In conclusion, skin fibroblasts of subjects with an impaired GSH synthesis showed an altered proteome reaction in response to oxidative stress. Furthermore, the study corroborates the use of fibroblasts as an additional mean to study vulnerability factors of psychiatric diseases.

Disrupting the EMMPRIN (CD147)-cyclophilin A interaction reduces infarct size and preserves systolic function after myocardial ischemia and reperfusion.

Objective-Inflammation and proteolysis crucially contribute to myocardial ischemia and reperfusion injury. The extracellular matrix metalloproteinase inducer EMMPRIN (CD147) and its ligand cyclophilin A (CyPA) may be involved in both processes. The aim of the study was to characterize the role of the CD147 and CyPA interplay in myocardial ischemia/reperfusion (I/R) injury.Methods and Results-Immunohistochemistry showed enhanced expression of CD147 and CyPA in myocardial sections from human autopsies of patients who had died from acute myocardial infarction and from mice at 24 hours after I/R. At 24 hours and 7 days after I/R, the infarct size was reduced in CD147(+/-) mice vs CD147(+/+) mice (C57Bl/6), in mice (C57Bl/6) treated with monoclonal antibody anti-CD147 vs control monoclonal antibody, and in CyPA(-/-) mice vs CyPA(+/+) mice (129S6/SvEv), all of which are associated with reduced monocyte and neutrophil recruitment at 24 hours and with a preserved systolic function at 7 days. The combination of CyPA(-/-) mice with anti-CD147 treatment did not yield further protection compared with either inhibition strategy alone. In vitro, treatment with CyPA induced monocyte chemotaxis in a CD147-and phosphatidylinositol 3-kinase-dependent manner and induced monocyte rolling and adhesion to endothelium (human umbilical vein endothelial cells) under flow in a CD147-dependent manner.Conclusion-CD147 and its ligand CyPA are inflammatory mediators after myocardial ischemia and reperfusion and represent potential targets to prevent myocardial I/R injury.

Splanchnic metabolism of nutrients and hormones in steers fed alfalfa under conditions of increased absorption of ammonia and L-arginine supply across the portal-drained viscera

Effects of increased ammonia and/or arginine absorption on net splanchnic (portal-drained viscera [PDV] plus liver) metabolism of nonnitrogenous nutrients and hormones in cattle were examined. Six Hereford x Angus steers (501 +/- 1 kg BW) prepared with vascular catheters for measurements of net flux across the splanchnic bed were fed a 75% alfalfa:25% (as-fed basis) corn and soybean meal diet (0.523 MJ of ME/[kg BW(0.75.)d]) every 2 h without (27.0 g of N/kg of DM) and. with 20 g of urea/kg of DM (35.7 g of N/kg of DM) in a split-plot design. Net flux measurements were made immediately before and after a 72-h mesenteric vein infusion Of L-arginine (15 mmol/h). There were no treatment effects on PDV or hepatic 02 consumption. Dietary urea had no effect on splanchnic metabolism of glucose or L-lactate, but arginine infusion decreased net hepatic removal Of L-lactate when urea was fed (P < 0.01). Net PDV appearance of n-butyrate was increased by arginine infusion (P < 0.07), and both dietary urea (P < 0.09) and arginine infusion (P < 0.05) increased net hepatic removal of n-butyrate. Dietary urea also increased total splanchnic acetate output (P < 0.06), tended to increase arterial glucagon concentration (P < 0.11), and decreased arterial ST concentration (P < 0.03). Arginine infusion increased arterial concentration (P < 0.07) and net PDV release (P < 0.10) and tended to increase hepatic removal (P < 0.11) of insulin, as well as arterial concentration (P < 0.01) and total splanchnic output (P < 0.01) of glucagon. Despite changes in splanchnic N metabolism, increased ammonia and arginine absorption had little measurable effect on splanchnic metabolism of glucose and other nonnitrogenous components of splanchnic energy metabolism.

Gut microbiota modulate the metabolism of brown adipose tissue in mice

A two by two experimental study has been designed to determine the effect of gut microbiota on energy metabolism in mouse models. The metabolic phenotype of germ-free (GF, n = 20) and conventional (n = 20) mice was characterized using a NMR spectroscopy-based metabolic profiling approach, with a focus on sexual dimorphism (20 males, 20 females) and energy metabolism in urine, plasma, liver, and brown adipose tissue (BAT). Physiological data of age-matched GF and conventional mice showed that male animals had a higher weight than females in both groups. In addition, conventional males had a significantly higher total body fat content (TBFC) compared to conventional females, whereas this sexual dimorphism disappeared in GF animals (i.e., male GF mice had a TBFC similar to those of conventional and GF females). Profiling of BAT hydrophilic extracts revealed that sexual dimorphism in normal mice was absent in GF animals, which also displayed lower BAT lactate levels and higher levels of (D)-3-hydroxybutyrate in liver, plasma, and BAT, together with lower circulating levels of VLDL. These data indicate that the gut microbiota modulate the lipid metabolism in BAT, as the absence of gut microbiota stimulated both hepatic and BAT lipolysis while inhibiting lipogenesis. We also demonstrated that (1)H NMR metabolic profiles of BAT were excellent predictors of BW and TBFC, indicating the potential of BAT to fight against obesity.

Top single nucleotide polymorphisms affecting carbohydrate metabolism in Metabolic Syndrome: from the LIPGENE study

Rationale:Metabolic Syndrome (MetS) is a high prevalence condition characterized by altered energy metabolism, insulin resistance and elevated cardiovascular risk.Objectives:Although many individual single nucleotide polymorphisms (SNPs) have been linked to certain MetS features, there are few studies analyzing the influence of SNPs on carbohydrate metabolism in MetS.Methods:904 SNPs (tag SNPs and functional SNPs) were tested for influence in eight fasting and dynamic markers of carbohydrate metabolism, performing an intravenous glucose tolerance test in 450 participants of the LIPGENE study.Findings:From 382 initial gene-phenotype associations between SNPs and any phenotypic variables, 61 (a 16 % of the pre-selected) remained significant after Bootstrapping. Top SNPs affecting glucose metabolism variables were as follows: fasting glucose: rs26125 (PPARGC1B); fasting insulin: rs4759277 (LRP1); C peptide: rs4759277 (LRP1); HOMA-IR: rs4759277 (LRP1); QUICKI: rs184003 (AGER); SI: rs7301876 (ABCC9), AIRg: rs290481 (TCF7L2) and DI: rs12691 (CEBPA).Conclusions:We describe here the top SNPs linked to phenotypic features in carbohydrate metabolism among aproximately 1000 candidate gene variations in fasting and postprandial samples of 450 patients with MetS from the LIPGENE study.

Short communication: plasma concentration of glucose-dependent insulinotropic polypeptide may regulate milk energy production in lactating dairy cows

In dairy cows, an increase in plasma concentration of glucose-dependent insulinotropic polypeptide (GIP) is associated with an increase in metabolizable energy intake, but the role of GIP in energy partitioning of dairy cattle is not certain. The objective of this study was to examine the relationship between plasma GIP concentrations and energy partitioning toward milk production. Four mid-lactation, primiparous, rumenfistulated Holstein-Friesian cows were fed a control diet of 55% forage and 45% concentrate [dry matter (DM) basis] in a 4 × 4 Latin square design with 4-wk periods. The 4 treatments were (1) control diet fed at 1000 and 1600 h, and (2) once-daily (1000 h) feeding, (3) twice daily (1000 and 1600 h) feeding, and (4) 4 times/d (1000, 1600, 2200 and 0400 h) feeding of the control diet plus 1 dose (1.75 kg on a DM basis at 0955 h) into the rumen of supplemental vegetable proteins (Amino Green; SCA NuTec Ltd., Thirsk, UK). Measurements of respiratory exchange and energy balance were obtained over 4 d during the last week of each period while cows were housed in open-circuit respiration chambers. Blood was collected from the jugular vein every 30 min for 12 h, using indwelling catheters, starting at 0800 h on d 20 of each period. Plasma GIP concentration was measured in samples pooled over each 5 consecutive blood samplings. The relationships between plasma GIP, DM intake, heat production, respiratory quotient, milk yield, and milk energy output were analyzed using linear correlation procedures, with metabolizable intake as a partial variant. Plasma GIP concentration was not correlated with heat production, or milk yield, but was positively correlated with milk energy yield (correlation coefficient = 0.67) and negatively correlated with RQ (correlation coefficient = −0.72). The correlations between GIP and RQ and milk energy output do not imply causality, but suggest that a role for GIP may exist in the regulation of energy metabolism in dairy cows.

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.

Effect of energy intake on performance and carcass composition of broiler chickens from two different genetic groups

In order to evaluate the effect of energy intake and broiler genotype on performance, carcass yield, and fat deposition, 600 one-day-old male chicks from two different genetic groups (AgRoss 308 - commercial line and PCLC - Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) non-improved line) were fed diets with different metabolizable energy level (2950, 3200 and 3450 kcal/kg). A completely randomized experimental design in a 2X3 factorial arrangement with four replications of 25 birds per treatment was applied. In order to ensure different energy intake among treatments within each strain, feed intake was daily adjusted by pair-feeding schemes. AgRoss 308 broilers had better performance and carcass yield, and presented lower abdominal fat deposition rate. In both genetic groups, the highest dietary energy level increased weight gain, heart relative weight, and fat deposition. However, it reduced the difference between AgRoss 308 and PCLC for feed conversion ratio and carcass protein deposition. These findings allow concluding that genetic improvement had a significant effect on broiler energy metabolism, and that the highest performance differences between genetic groups are found when low-energy intake is imposed.

Modeling energy utilization in broiler breeders, laying hens and broilers

The factorial approach has been used to partition the energy requirements into maintenance, growth, and production. The coefficients determined for these purposes can be used to elaborate energy requirement models. These models consider the body weight, weight gain, egg production, and environmental temperature to determine the energy requirements for poultry. Predicting daily energy requirement models can help to establish better and more profitable feeding programs for poultry. Studies were conducted at UNESP-Jaboticabal to determine metabolizable energy (ME) requirement models for broiler breeders, laying hens, and broilers. These models were evaluated in performance trials and provided good adjustments. Therefore, they could be used to establish nutritional programs. This review aims to outline the results found at UNESP studies and to show the application of models in nutritional programs for broiler breeders, laying hens, and broilers.

Energy expenditure during daily activities as measured by two motion sensors in patients with COPD

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

Ventricular Remodeling Induced by Tissue Vitamin A Deficiency in Rats

Background/Aims: Experimental studies suggest that vitamin A plays a role in regulating cardiac structure and function. We tested the hypothesis that cardiac vitamin A deficiency is associated with adverse myocardial remodeling in young adult rats. Methods: Two groups of young female rats, control (C - n = 29) and tissue vitamin A deficient (RVA - n = 31), were subjected to transthoracic echocardiography exam, isolated rat heart study and biochemical study. Results: The RVA rats showed a reduced total vitamin A concentration in both the liver and heart [vitamin A in heart, mu mol/kg (C = 0.95 +/- 0.44 and RVA = 0.24 +/- 0.16, p = 0.01)] with the same serum retinol levels (C = 0.73 +/- 0.29 mu mol/L e RVA = 0.62 +/- 0.17 mu mol/L, p = 0.34). The RVA rats showed higher left ventricular diameters and reduced systolic function. The RVA rats also demonstrated increased lipid hydroperoxide/total antioxidant capacity ratio and cardiac levels of IFN-gamma and TNF-alpha but not of metalloproteinase (MMP)-2 and -9 activity. on the other hand, the RVA rats had decreased levels of beta-hydroxyacylcoenzyme A dehydrogenase and lactate dehydrogenase. Conclusions: Tissue vitamin A deficiency stimulated cardiac remodeling and ventricular dysfunction. Additionally, the data support the involvement of oxidative stress, energy metabolism, and cytokine production in this remodeling process. Copyright (C) 2010 S. Karger AG, Basel

Energy system contributions in indoor rock climbing

The present study cross-sectionally investigated the influence of training status, route difficulty and upper body aerobic and anaerobic performance of climbers on the energetics of indoor rock climbing. Six elite climbers (EC) and seven recreational climbers ( RC) were submitted to the following laboratory tests: ( a) anthropometry, (b) upper body aerobic power, and ( c) upper body Wingate test. on another occasion, EC subjects climbed an easy, a moderate, and a difficult route, whereas RC subjects climbed only the easy route. The fractions of the aerobic (WAER), anaerobic alactic (W-PCR) and anaerobic lactic (W-[La(])-) systems were calculated based on oxygen uptake, the fast component of excess post-exercise oxygen uptake, and changes in net blood lactate, respectively. on the easy route, the metabolic cost was significantly lower in EC [ 40.3 ( 6.5) kJ] than in RC [60.1 ( 8.8) kJ] ( P < 0.05). The respective contributions of the WAER, WPCR, and W-[La(])- systems in EC were: easy route = 41.5 (8.1), 41.1 (11.4) and 17.4% (5.4), moderate route = 45.8 (8.4), 34.6 (7.1) and 21.9% (6.3), and difficult route = 41.9 (7.4), 35.8 (6.7) and 22.3% (7.2). The contributions of the WAER, WPCR, and W-[La(])- systems in RC subjects climbing an easy route were 39.7 (5.0), 34.0 (5.8), and 26.3% (3.8), respectively. These results indicate that the main energy systems required during indoor rock climbing are the aerobic and anaerobic alactic systems. In addition, climbing economy seems to be more important for the performance of these athletes than improved energy metabolism.

Effects of olive oil and its minor phenolic constituents on obesity-induced cardiac metabolic changes

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

Putative pyrophosphate phosphofructose 1-kinase genes identified in sugarcane may be getting energy from pyrophosphate

Pyrophosphate-dependent phosphofructokinase (PPi-PFK) has been detected in several types of plant cells, but the gene has not been reported in sugar cane. Using Citrux paradixi PPi-PFK gene (AF095520 and AF095521) sequences to search the sugar cane EST database, we have identified both the α and β subunits of this enzyme. The deduced amino acid sequences showed 76 and 80% similarity with the corresponding α and β subunits of C. paradisi. A high degree of similarity was also observed among the PFK β subunits when the alignment of the sugar cane sequences was compared to those of Ricinus communis and Solanum tuberosum, it appears that α and β are two distinct subunits; they were found at different concentrations in several sugar cane tissues. It remains to be determined if the different gene expression levels have some physiological importance and how they affect sucrose synthesis, export, and storage in vacuoles. A comparison between the amino acid sequences of β PFKs from a variety of organisms allowed us to identify the two critical Asp residues typical of this enzyme's activity site and the other binding sites; these residues are tightly conserved in all members of this protein family. Apparently, there are catalytic residues on the β subunit of the pyrophosphate-dependent enzyme.