Robust arm and leg muscle adaptation to training despite ACE inhibition: a randomized placebo-controlled trial.

PURPOSE: Angiotensin-converting enzyme (ACE) inhibitor treatment is widely applied, but the fact that plasma ACE activity is a potential determinant of training-induced local muscular adaptability is often neglected. Thus, we investigated the hypothesis that ACE inhibition modulates the response to systematic aerobic exercise training on leg and arm muscular adaptations. METHODS: Healthy, untrained, middle-aged participants (40 ± 7 yrs) completed a randomized, double-blinded, placebo-controlled trial. Participants were randomized to placebo (PLA: CaCO3) or ACE inhibitor (ACEi: enalapril) for 8 weeks and completed a supervised, high-intensity exercise training program. Muscular characteristics in the leg and arm were extensively evaluated pre and post-intervention. RESULTS: Forty-eight participants (nACEi = 23, nPLA = 25) completed the trial. Exercise training compliance was above 99%. After training, citrate synthase, 3-hydroxyacyl-CoA dehydrogenase and phosphofructokinase maximal activity were increased in m. vastus lateralis in both groups (all P < 0.05) without statistical differences between them (all time × treatment P > 0.05). In m. deltoideus, citrate synthase maximal activity was upregulated to a greater extent (time × treatment P < 0.05) in PLA (51 [33;69] %) than in ACEi (28 [13;43] %), but the change in 3-hydroxyacyl-CoA dehydrogenase and phosphofructokinase maximal activity was similar between groups. Finally, the training-induced changes in the platelet endothelial cell adhesion molecule-1 protein abundance, a marker of capillary density, were similar in both groups in m. vastus lateralis and m. deltoideus. CONCLUSION: Eight weeks of high-intensity whole-body exercise training improves markers of skeletal muscle mitochondrial oxidative capacity, glycolytic capacity and angiogenesis, with no overall effect of pharmacological ACE inhibition in healthy adults.

Hadh deficiency induced oligoasthenoteratozoospermia through the TNF-α/Bcl-2 pathway in male mice.

The process of spermatogenesis is a complex and delicate process that is still not fully understood. In this study, we examined the role of fatty acid oxidase 3-hydroxy acyl CoA dehydrogenase (HADH) in maintaining normal spermatogenesis in mice. In male mice, ablation of the Hadh gene using CRISPR/Cas9 technology arrested spermatocyte meiosis, increased multinucleated giant germ cells and vacuoles in seminiferous tubules, and accompanied with acrosomal dysplasia. Hadh-/- male mice showed the typical features of oligoasthenoteratozoospermia (OAT), including decreased sperm concentration and motility and increased sperm abnormalities. Next, we explored the molecular events in the testes of the mutant mice. We found fatty acids accumulated in the testis of Hadh-/- mice. And also, inflammatory factors TNF-α, IL-1ß, and IL-6 were significantly increased, apoptosis-related protein Bcl-2 was decreased, and Bax and cleaved-Caspase3 were increased in Hadh-/- male mice testis. After using etanercept, a specific inhibitor of TNF-α, testis injury caused by Hadh knockout was significantly alleviated, the sperm quality and motility were improved, and germ cell apoptosis was reduced. So our study demonstrated that Hadh deletion caused an increase in fatty acids. The accumulated fatty acids further induced testicular inflammation and germ cell apoptosis through the TNF-α/Bcl-2 signaling pathway, finally resulting in OAT in the Hadh-/- mice. Inhibiting TNF-α may be used as a new treatment approach for testicular inflammation and OAT.

Functional analysis of HADH c.99C>G shows that the variant causes the proliferation of pancreatic islets and leu-sensitive hyperinsulinaemia.

A novel missense variant (NM_005327.7: c.99C>G, p.Ile33Met) was discovered in 3-hydroxyacyl-CoA dehydrogenase (HADH), which is involved in congenital hyperinsulinism (CHI). This variant may be damaging or deleterious, as assessed using protein prediction software. This study aimed at the impact of this variant on islets and if it caused the leu-sensitive insulin secretion. The adenoassociated virus containing the HADH missense variant (p.Ile33Met), wild-type (WT) HADH or empty vector (EV) was constructed, and the rats were infected with it. Three weeks after the transfection, 15 rats were dissected to observe the effect of the variant on the islet tissue. Then we treated the remaining rats with leucine or sodium carboxymethyl cellulose (CMC-Na) by gavage and drew blood from the rat tail vein to detect the variations in blood glucose, serum insulin and serum glucagon. Further, we dissected the rats to observe the fluctuation of insulin and glucagon contents in pancreatic islets under the combined action of leucine and p.Ile33Met. Insulin and glucagon were observed in the islet tissue under an inverted fluorescence microscope, serum insulin and glucagon were detected by ELISA, and the blood glucose value was determined using a Roche glucometer. The positive area and average gray value of islet fluorescence pictures were analysed using the software Image J (USA). Rats expressing p.Ile33Met showed significantly higher insulin and glucagon content, as well as the islet area, compared to WT and EV rats. Moreover, after intragastric administration of leucine, the serum insulin content of the variant rats increased but the blood sugar level decreased significantly. Meanwhile, there was an appreciable decrease in the insulin content in rat pancreatic islet tissues. Our results suggest that the variant NM_005327.7: c.99C>G promotes the proliferation of pancreatic islets, enhances the secretion of insulin, and induces leu-sensitive hyperinsulinaemia.

Salinomycin biosynthesis reversely regulates the ß-oxidation pathway in Streptomyces albus by carrying a 3-hydroxyacyl-CoA dehydrogenase gene in its biosynthetic gene cluster.

Streptomyces is well known for synthesis of many biologically active secondary metabolites, such as polyketides and non-ribosomal peptides. Understanding the coupling mechanisms of primary and secondary metabolism can help develop strategies to improve secondary metabolite production in Streptomyces. In this work, Streptomyces albus ZD11, an oil-preferring industrial Streptomyces strain, was proved to have a remarkable capability to generate abundant acyl-CoA precursors for salinomycin biosynthesis with the aid of its enhanced ß-oxidation pathway. It was found that the salinomycin biosynthetic gene cluster contains a predicted 3-hydroxyacyl-CoA dehydrogenase (FadB3), which is the third enzyme of ß-oxidation cycle. Deletion of fadB3 significantly reduced the production of salinomycin. A variety of experimental evidences showed that FadB3 was mainly involved in the ß-oxidation pathway rather than ethylmalonyl-CoA biosynthesis and played a very important role in regulating the rate of ß-oxidation in S. albus ZD11. Our findings elucidate an interesting coupling mechanism by which a PKS biosynthetic gene cluster could regulate the ß-oxidation pathway by carrying ß-oxidation genes, enabling Streptomyces to efficiently synthesize target polyketides and economically utilize environmental nutrients.

Crystal structure of multi-functional enzyme FadB from Cupriavidus necator: Non-formation of FadAB complex.

Cupriavidus necator H16 is a gram-negative chemolithoautotrophic bacterium that has been extensively studied for biosynthesis and biodegradation of polyhydroxyalkanoate (PHA) plastics. To improve our understanding of fatty acid metabolism for PHA production, we determined the crystal structure of multi-functional enoyl-CoA hydratase from Cupriavidus necator H16 (CnFadB). The predicted model of CnFadB created by AlphaFold was used to solve the phase problem during determination of the crystal structure of the protein. The CnFadB structure consists of two distinctive domains, an N-terminal enol-CoA hydratase (ECH) domain and a C-terminal 3-hydroxyacyl-CoA dehydrogenase (HAD) domain, and the substrate- and cofactor-binding modes of these two functional domains were identified. Unlike other known FadB enzymes that exist as dimers complexed with FadA, CnFadB functions as a monomer without forming a complex with CnFadA. Small angle X-ray scattering (SAXS) measurement further proved that CnFadB exists as a monomer in solution. The non-sequential action of FadA and FadB in C. necator appears to affect ß-oxidation and PHA synthesis/degradation.

iTRAQ-facilitated proteomic analysis of Bacillus cereus via degradation of malachite green.

The wide use of malachite green (MG) as a dye has caused substantial concern owing to its toxicity. Bacillus cereus can against the toxic effect of MG and efficiently decolourise it. However, detailed information regarding its underlying adaptation and degradation mechanisms based on proteomic data is scarce. In this study, the isobaric tags for relative and absolute quantitation (iTRAQ)-facilitated quantitative method was applied to analyse the molecular mechanisms by which B. cereus degrades MG. Based on this analysis, 209 upregulated proteins and 198 downregulated proteins were identified with a false discovery rate of 1% or less during MG biodegradation. Gene ontology and KEGG analysis determined that the differentially expressed proteins were enriched in metabolic processes, catalytic activity, antioxidant activity, and responses to stimuli. Furthermore, real-time qPCR was utilised to further confirm the regulated proteins involved in benzoate degradation. The proteins BCE_4076 (Acetyl-CoA acetyltransferase), BCE_5143 (Acetyl-CoA acetyltransferase), BCE_5144 (3-hydroxyacyl-CoA dehydrogenase), BCE_4651 (Enoyl-CoA hydratase), and BCE_5474 (3-hydroxyacyl-CoA dehydrogenase) involved in the benzoate degradation pathway may play an important role in the biodegradation of MG by B. cereus. The results of this study not only provide a comprehensive view of proteomic changes in B. cereus upon MG loading but also shed light on the mechanism underlying MG biodegradation by B. cereus.

Distribution of concurrent training sessions does not impact endurance adaptation.

OBJECTIVES: Optimized concurrent training regimes are warranted in physical training of military-, law enforcement- and rescue-personnel. This study investigated if four 15-min endurance training sessions weekly improve aerobic capacity and performance more than one 60-min endurance session weekly during the initial phase of a Basic Military Training program. DESIGN: A randomized training intervention study with functional and physiological tests before and after the intervention. METHODS: Military conscripts (n=290) were randomly allocated to three groups completing 9 weeks training. Weekly training consisted of four endurance and four strength training sessions lasting 15min each ('Micro-training': MIC); one strength and one endurance session lasting 60min each ('Classical-training': CLA) or two 60min sessions of standard military training ('Control-training': CON). RESULTS: Both 12-min (∼7-10%) and shuttle run performance (∼35-42%) improved (P≤0.001) similarly in all groups. Likewise, functional 2-min maximal repetition exercise capacity increased (P≤0.05) similarly in all groups (Lunges ∼17-24 %; PushUp ∼10-20%; AbdominalFlexions∼21-23%). Peak oxygen uptake changes depended on group (P≤0.05) with increases (P≤0.01) in MIC (7±7%, n=23) and CON (12±18%, n=17) and no changes in CLA. Maximal m. vastus lateralis citrate synthase activity decreased 14±26% (P≤0.001, n=18) in CLA. Likewise, maximal m. vastus lateralis 3-hydroxyacyl-CoA dehydrogenase activity decreased 8±17% in MIC (n=28) and 14±24% in CLA (n=18). CONCLUSIONS: Four 15-min endurance training sessions weekly improves running performance and strength-endurance similarly to one 60min session. Peak oxygen uptake only increases with more than one endurance session weekly and leg muscle oxidative capacity appears reduced after basic military training.

Effects of lactate administration on mitochondrial enzyme activity and monocarboxylate transporters in mouse skeletal muscle.

Growing evidence shows that lactate is not merely an intermediate metabolite, but also a potential signaling molecule. However, whether daily lactate administration induces physiological adaptations in skeletal muscle remains to be elucidated. In this study, we first investigated the effects of daily lactate administration (equivalent to 1 g/kg of body weight) for 3 weeks on mitochondrial adaptations in skeletal muscle. We demonstrated that 3-week lactate administration increased mitochondrial enzyme activity (citrate synthase, 3-hydroxyacyl CoA dehydrogenase, and cytochrome c oxidase) in the plantaris muscle, but not in the soleus muscle. MCT1 and MCT4 protein contents were not different after 3-week lactate administration. Next, we examined whether lactate administration enhances training-induced adaptations in skeletal muscle. Lactate administration prior to endurance exercise training (treadmill running, 20 m/min, 60 min/day), which increased blood lactate concentration during exercise, enhanced training-induced mitochondrial enzyme activity in the skeletal muscle after 3 weeks. MCT protein content and blood lactate removal were not different after 3-week lactate administration with exercise training compared to exercise training alone. In a single bout experiment, lactate administration did not change the phosphorylation state of AMPK, ACC, p38 MAPK, and CaMKII in skeletal muscle. Our results suggest that lactate can be a key factor for exercise-induced mitochondrial adaptations, and that the efficacy of high-intensity training is, at least partly, attributed to elevated blood lactate concentration.

Effects of exercise training with short-duration intermittent hypoxia on endurance performance and muscle metabolism in well-trained mice.

The author previously reported that short-duration intermittent hypoxia had additive effects on improvements in endurance capacity by enhancing fatty acid metabolism. The present study was designed to investigate the effects of short-duration intermittent hypoxia on endurance capacity, metabolic enzyme activity, and protein levels associated with mitochondrial biogenesis in well-trained mice. Mice in the training group were housed in a cage with a running wheel for 7 weeks from 5 weeks old. Voluntary running markedly increased maximal work values by 5.0-fold. Trained mice were then subjected to either endurance treadmill training (ET) for 60 min or hybrid training (HT, ET for 30 min followed by sprint interval exercise (5-sec run-10-sec rest) for 30 min) with (H-ET or H-HT) or without (ET or HT) short-duration intermittent hypoxia (4 cycles of 12-13% O2 for 15 min and 20.9% O2 for 10 min) for 4 weeks. Maximal endurance capacity was markedly greater in the H-ET and H-HT than ET and HT groups, respectively. H-ET and H-HT increased activity levels of 3-hydroxyacyl-CoA-dehydrogenase in oxidative muscle portion and pyruvate dehydrogenase complex in glycolytic muscle portion. These activity levels were significantly correlated with maximal endurance capacity. Protein levels of dynamin-related protein-1 were increased more by H-ET and H-HT than by ET and HT, but were not significantly correlated with maximal work. These results suggest that intermittent hypoxic exposure has beneficial effects on endurance and hybrid training to improve the endurance capacity via improving fatty acid and pyruvate oxidation in highly trained mice.

1064 nm Nd:YAG laser light affects transmembrane mitochondria respiratory chain complexes.

Photobiomodulation (PBM) is a non-plant-cell manipulation through a transfer of energy by means of light sources at the non-ablative or thermal intensity. Authors showed that cytochrome-c-oxidase (complex IV) is the specific chromophore's target of PBM at the red (600-700 nm) and NIR (760-900 nm) wavelength regions. Recently, it was suggested that the infrared region of the spectrum could influence other chromospheres, despite the interaction by wavelengths higher than 900 nm with mitochondrial chromophores was not clearly demonstrated. We characterized the interaction between mitochondria respiratory chain, malate dehydrogenase, a key enzyme of Krebs cycle, and 3-hydroxyacyl-CoA dehydrogenase, an enzyme involved in the ß-oxidation (two mitochondrial matrix enzymes) with the 1064 nm Nd:YAG (100mps and 10 Hz frequency mode) irradiated at the average power density of 0.50, 0.75, 1.00, 1.25 and 1.50 W/cm2 to generate the respective fluences of 30, 45, 60, 75 and 90 J/cm2 . Our results show the effect of laser light on the transmembrane mitochondrial complexes I, III, IV and V (adenosine triphosphate synthase) (window effects), but not on the extrinsic mitochondrial membrane complex II and mitochondria matrix enzymes. The effect is not due to macroscopical thermal change. An interaction of this wavelength with the Fe-S proteins and Cu-centers of respiratory complexes and with the water molecules could be supposed.

The p75NTR-mediated effect of nerve growth factor in L6C5 myogenic cells.

OBJECTIVE: During muscle development or regeneration, myocytes produce nerve growth factor (NGF) as well as its tyrosine-kinase and p75-neurotrophin (p75NTR) receptors. It has been published that the p75NTR receptor could represent a key regulator of NGF-mediated myoprotective effect on satellite cells, but the precise function of NGF/p75 signaling pathway on myogenic cell proliferation, survival and differentiation remains fragmented and controversial. Here, we verified the role of NGF in the growth, survival and differentiation of p75NTR-expressing L6C5 myogenic cells, specifically inquiring for the putative involvement of the nuclear factor κB (NFκB) and the small heat shock proteins (sHSPs) αB-crystallin and Hsp27 in these processes. RESULTS: Although NGF was not effective in modulating myogenic cell growth or survival in both standard or stress conditions, we demonstrated for the first time that, under serum deprivation, NGF sustained the activity of some key enzymes involved in energy metabolism. Moreover, we confirmed that NGF promotes myogenic fusion and expression of the structural protein myosin heavy chain while modulating NFκB activation and the content of sHSPs correlated with the differentiation process. We conclude that p75NTR is sufficient to mediate the modulation of L6C5 myogenic differentiation by NGF in term of structural, metabolic and functional changes.

Exercise with weight loss improves adipose tissue and skeletal muscle markers of fatty acid metabolism in postmenopausal women.

OBJECTIVE: The effects of 6-month weight loss (WL) versus aerobic exercise training (AEX)+WL on fat and skeletal muscle markers of fatty acid metabolism were determined in normal (NGT) and impaired (IGT) glucose tolerant African-American and Caucasian postmenopausal women with overweight/obesity. METHODS: Fat (gluteal and abdominal) lipoprotein lipase (LPL), skeletal muscle LPL, acyl-CoA synthase (ACS), ß-hydroxacyl-CoA dehydrogenase, carnitine palmitoyltransferase (CPT-1), and citrate synthase (CS) activities were measured at baseline (n = 104) and before and after WL (n = 34) and AEX+WL (n = 37). RESULTS: After controlling for age and race, muscle LPL and CPT-1 were lower in IGT, and the ratios of fat/muscle LPL activity were higher in IGT compared to NGT. Muscle LPL was related to insulin sensitivity (M value) and inversely related to G120 , fasting insulin, and homeostatic model assessment of insulin resistance. AEX+WL decreased abdominal fat LPL and increased muscle LPL, ACS, and CS. The ratios of fat/muscle LPL decreased after AEX+WL. The change in VO2 max was related to the changes in LPL, ACS, and CS and inversely related to the changes in fat/muscle LPL activity ratios. CONCLUSIONS: Six-month AEX+WL, and not WL alone, is capable of enhancing skeletal muscle fatty acid metabolism in postmenopausal African-American and Caucasian women with NGT, IGT, and overweight/obesity.

Effects of immobilization and aerobic training on proteins related to intramuscular substrate storage and metabolism in young and older men.

PURPOSE: Aging and inactivity lead to skeletal muscle metabolic inflexibility, but the underlying molecular mechanisms are not entirely elucidated. Therefore, we investigated how muscle lipid and glycogen stores and major regulatory proteins were affected by short-term immobilization followed by aerobic training in young and older men. METHODS: 17 young (23 ± 1 years, 24 ± 1 kg m(-2), and 20 ± 2% body fat) and 15 older men (68 ± 1 years; 27 ± 1 kg m(-2), and 29 ± 2% body fat) underwent 2 weeks' one leg immobilization followed by 6 weeks' cycle training. Biopsies were obtained from m. vastus lateralis just before immobilization (at inclusion), after immobilization, and the after 6 weeks' training. The biopsies were analyzed for muscle substrates; muscle perilipin protein (PLIN), glycogen synthase (GS), synaptosomal-associated protein of 23 kDa (SNAP23) protein content, and muscle 3-hydroxyacyl-CoA dehydrogenase (HAD) activity RESULTS: The older men had higher intramuscular triglyceride (IMTG) (73 %) and Glycogen (16%) levels compared to the young men, and IMTG tended to increase with immobilization. PLIN2 and 3 protein content increased with immobilization in the older men only. The young men had higher GS (74%) protein compared to the older men. Immobilization decreased and training restored HAD activity, GS and SNAP23 protein content in young and older men. CONCLUSION: Evidence of age-related metabolic inflexibility is presented, seen as body fat and IMTG accumulation. The question arises as to whether IMTG accumulation in the older men is caused by or leading to the increase in PLIN2 and 3 protein content. Training decreased body fat and IMTG levels in both young and older men; hence, training should be prioritized to reduce the detrimental effect of aging on metabolism.

Grape seed procyanidins administered at physiological doses to rats during pregnancy and lactation promote lipid oxidation and up-regulate AMPK in the muscle of male offspring in adulthood.

The aim of the present study was to test whether the administration of a grape seed procyanidin extract (GSPE) during pregnancy and lactation, at doses extrapolated to human consumption, programs male offspring toward improved metabolism in adulthood. For this purpose, female rats were fed a normal-fat diet (NFD) and treated with either GSPE (25 mg kg(-1) of body weight/day) or vehicle during gestation and lactation. The metabolic programming effects of GSPE were evaluated in the male offspring fed NFD from 30 to 170 days of life. No changes were observed in body weight, adiposity, circulating lipid profile and insulin sensitivity between the offspring of dams treated with GSPE (STD-GSPE group) and their counterparts (STD-veh). However, the STD-GSPE offspring had lower circulating levels of C-reactive protein and lower respiratory quotient values, shifting whole-body energy catabolism from carbohydrate to fat oxidation. Furthermore, the STD-GSPE animals also exhibited increased levels of total and phosphorylated AMP-activated protein kinase (AMPK) and an over-expression of the mRNA levels of key genes related to fatty acid uptake (Fatp1 and CD36) and ß-oxidation (pparα and had) in skeletal muscle. Our results indicate that GSPE programs healthy male offspring towards a better circulating inflammatory profile and greater lipid utilisation in adulthood. The metabolic programming effects of GSPE that are related to the enhancement of fatty acid oxidation in skeletal muscle seem to be mediated, at least in part, by AMPK. These findings could be of relevance in the prevention of pathologies associated to lifestyle and aging, such as obesity and insulin resistance.

Blocking phosphatidylcholine utilization in Pseudomonas aeruginosa, via mutagenesis of fatty acid, glycerol and choline degradation pathways, confirms the importance of this nutrient source in vivo.

Pseudomonas aeruginosa can grow to very high-cell-density (HCD) during infection of the cystic fibrosis (CF) lung. Phosphatidylcholine (PC), the major component of lung surfactant, has been hypothesized to support HCD growth of P. aeruginosa in vivo. The phosphorylcholine headgroup, a glycerol molecule, and two long-chain fatty acids (FAs) are released by enzymatic cleavage of PC by bacterial phospholipase C and lipases. Three different bacterial pathways, the choline, glycerol, and fatty acid degradation pathways, are then involved in the degradation of these PC components. Here, we identified five potential FA degradation (Fad) related fadBA-operons (fadBA1-5, each encoding 3-hydroxyacyl-CoA dehydrogenase and acyl-CoA thiolase). Through mutagenesis and growth analyses, we showed that three (fadBA145) of the five fadBA-operons are dominant in medium-chain and long-chain Fad. The triple fadBA145 mutant also showed reduced ability to degrade PC in vitro. We have previously shown that by partially blocking Fad, via mutagenesis of fadBA5 and fadDs, we could significantly reduce the ability of P. aeruginosa to replicate on FA and PC in vitro, as well as in the mouse lung. However, no studies have assessed the ability of mutants, defective in choline and/or glycerol degradation in conjunction with Fad, to grow on PC or in vivo. Hence, we constructed additional mutants (ΔfadBA145ΔglpD, ΔfadBA145ΔbetAB, and ΔfadBA145ΔbetABΔglpD) significantly defective in the ability to degrade FA, choline, and glycerol and, therefore, PC. The analysis of these mutants in the BALB/c mouse lung infection model showed significant inability to utilize PC in vitro, resulted in decreased replication fitness and competitiveness in vivo compared to the complement strain, although there was little to no variation in typical virulence factor production (e.g., hemolysin, lipase, and protease levels). This further supports the hypothesis that lung surfactant PC serves as an important nutrient for P. aeruginosa during CF lung infection.

Increased ß-oxidation with improved glucose uptake capacity in adipose tissue from obese after weight loss and maintenance.

OBJECTIVE: We investigated protein markers for pathways of the fatty acids (FAs) and glucose metabolism in human adipose tissue after a weight loss program by calorie restriction. METHODS: Overweight/obese subjects underwent an intervention of 5 weeks of a very low-calorie diet followed by a 3-week weight maintenance diet. Abdominal subcutaneous adipose tissue biopsies were sampled before and after the intervention. Seventeen target proteins as markers of metabolic pathways for the uptake and handling of FAs and glucose were quantified by Western blotting and 11 were retrieved from previous proteomics work. Correlation coefficients were calculated among changes of these proteins. RESULTS: Short-chain 3-hydroxyacyl-CoA dehydrogenase, catalase, fatty acid translocase, fatty acid transporter protein 3, adipose triglyceride lipase, fatty acid-binding protein 4, aldolase-C, tubulin-ß-5, and annexin A2 changed significantly, and lipoprotein lipase, perilipin 1, and hormone-sensitive lipase tended to change. On an average, increased glucose transporter type 4 translocation was observed, supported by a consistent increase of tyr-24 phosphorylated annexin A2. CONCLUSIONS: Our findings suggest that after weight loss by calorie restriction and a short period of maintenance, adipose tissue has an increased capacity for glucose uptake, and lipid mobilization and oxidation. Such metabolic profile may relate to the health benefit of weight loss.

Metabolic adaptation of two pig muscles to cold rearing conditions.

Cold environment represents an external stress modulating animal growth and energy use. At muscle level, adaptation to cold conditions potentially involves energy homeostasis regulation gauged by the adenosine monophosphate (AMP)-activated protein kinase (AMPK). Our study aimed at evaluating the bare effects of short- and long-term cold exposure on growth performance, carcass traits, and metabolic characteristics of the oxidative semispinalis (SS) muscle and glycolytic LM and to evaluate the reversibility of short-term effects, with a special emphasis on AMPK activity. A total of 84 pigs fed ad libitum and individually housed were submitted after weaning to either Cold (from 23 ± 1 to 15 ± 3°C) or thermoneutral (T; from 28 ± 1 to 23 ± 1°C) temperature up to 24.7 ± 1.6 kg BW (25 BW). Twelve Cold and 12 T piglets were then slaughtered the same day. Eighteen remaining Cold piglets were reared at 12 ± 2°C (CC) whereas 18 Cold and 24 T piglets were reared at 23 ± 4°C (CT and TT, respectively) and slaughtered at 114.3 ± 5.9 kg (115 BW). The LM and SS samples were analyzed to determine glycolytic potential (GP), activities of lactate dehydrogenase (LDH), citrate synthase (CS), and ß-hydroxy-acyl-CoA dehydrogenase (HAD) and to quantify AMPK phosphorylation Threonine 172 phosphorylated form of AMPK α1+α2 isoforms (pAMPK) / total AMPK α1+α2 isoforms (AMPK). Despite a greater ADFI (P < 0.001), Cold piglets exhibited less ADG (P < 0.001) and body fatness (P < 0.03) attesting an acute adaptation to a short-term cold exposure. A long-term cold adaptation evaluated on 115 BW pigs increased ADFI (P < 0.001) but did not influence ADG and carcass lean meat content. Cold environment influenced the dynamic of muscle metabolism in a muscle type dependent manner corresponding to an earlier and acute adaptation in SS from 8 kg onward and a belated adaptation in LM from 25 BW to 115 BW. Cold exposure was associated with a progressive increase of muscle oxidative capacity, first in the SS with greater HAD (P = 0.002) and CS activities (P = 0.03) at 25 BW and then both in SS and LM at 115 BW (P < 0.001). Conversely, in LM of CC pigs, increased GP (P < 0.001) and LDH activity (P = 0.03) were observed in addition to increased CS and HAD activities, highlighting the ability of LM to increase both its glycolytic and oxidative metabolism and to diversify its energy substrates. Pigs from CC group exhibited also less pAMPK/AMPK (P < 0.01) specifically in red SS muscle, denoting a reduced metabolic stress of this muscle after a long-term cold adaptation.