Lipid Oxidation Capacity Research Articles

The Role of Exerkines in Obesity-Induced Disruption of Mitochondrial Homeostasis in Thermogenic Fat.

There is a notable correlation between mitochondrial homeostasis and metabolic disruption. In this review, we report that obesity-induced disruption of mitochondrial homeostasis adversely affects lipid metabolism, adipocyte differentiation, oxidative capacity, inflammation, insulin sensitivity, and thermogenesis in thermogenic fat. Elevating mitochondrial homeostasis in thermogenic fat emerges as a promising avenue for developing treatments for metabolic diseases, including enhanced mitochondrial function, mitophagy, mitochondrial uncoupling, and mitochondrial biogenesis. The exerkines (e.g., myokines, adipokines, batokines) released during exercise have the potential to ameliorate mitochondrial homeostasis, improve glucose and lipid metabolism, and stimulate fat browning and thermogenesis as a defense against obesity-associated metabolic diseases. This comprehensive review focuses on the manifold benefits of exercise-induced exerkines, particularly emphasizing their influence on mitochondrial homeostasis and fat thermogenesis in the context of metabolic disorders associated with obesity.

Concerted regulation of skeletal muscle metabolism and contractile properties by the orphan nuclear receptor Nr2f6.

The maintenance of skeletal muscle plasticity upon changes in the environment, nutrient supply, and exercise depends on regulatory mechanisms that couple structural and metabolic adaptations. The mechanisms that interconnect both processes at the transcriptional level remain underexplored. Nr2f6, a nuclear receptor, regulates metabolism and cell differentiation in peripheral tissues. However, its role in the skeletal muscle is still elusive. Here, we aimed to investigate the effects of Nr2f6 modulation on muscle biology in vivo and in vitro. Global RNA-seq was performed in Nr2f6 knockdown C2C12 myocytes (N=4-5). Molecular and metabolic assays and proliferation experiments were performed using stable Nr2f6 knockdown and Nr2f6 overexpression C2C12 cell lines (N=3-6). Nr2f6 content was evaluated in lipid overload models in vitro and in vivo (N=3-6). In vivo experiments included Nr2f6 overexpression in mouse tibialis anterior muscle, followed by gene array transcriptomics and molecular assays (N=4), ex vivo contractility experiments (N=5), and histological analysis (N=7). The conservation of Nr2f6 depletion effects was confirmed in primary skeletal muscle cells of humans and mice. Nr2f6 knockdown upregulated genes associated with muscle differentiation, metabolism, and contraction, while cell cycle-related genes were downregulated. In human skeletal muscle cells, Nr2f6 knockdown significantly increased the expression of myosin heavy chain genes (two-fold to three-fold) and siRNA-mediated depletion of Nr2f6 increased maximal C2C12 myocyte's lipid oxidative capacity by 75% and protected against lipid-induced cell death. Nr2f6 content decreased by 40% in lipid-overloaded myotubes and by 50% in the skeletal muscle of mice fed a high-fat diet. Nr2f6 overexpression in mice resulted in an atrophic and hypoplastic state, characterized by a significant reduction in muscle mass (15%) and myofibre content (18%), followed by an impairment (50%) in force production. These functional phenotypes were accompanied by the establishment of an inflammation-like molecular signature and a decrease in the expression of genes involved in muscle contractility and oxidative metabolism, which was associated with the repression of the uncoupling protein 3 (20%) and PGC-1α (30%) promoters activity following Nr2f6 overexpression in vitro. Additionally, Nr2f6 regulated core components of the cell division machinery, effectively decoupling muscle cell proliferation from differentiation. Our findings reveal a novel role for Nr2f6 as a molecular transducer that plays a crucial role in maintaining the balance between skeletal muscle contractile function and oxidative capacity. These results have significant implications for the development of potential therapeutic strategies for metabolic diseases and myopathies.

Alterations of Photosynthetic and Oxidative Processes Influenced by the Presence of Different Zinc and Cadmium Concentrations in Maize Seedlings: Transition from Essential to Toxic Functions.

The study examined the impact of varying the concentrations of zinc (Zn) on plant responses, particularly on photosynthetic and oxidative metabolic processes. This investigation aimed to distinguish between the beneficial and harmful effects of Zn on plants, highlighting significant nutrient supply concerns. The investigation methods were centered around non-invasive methods, such as biophoton emission (delayed fluorescence-DF, ultra-weak bioluminescence-UWLE), fluorescence induction (Fv/Fm) measurements, chlorophyll content estimation (SPAD) and vegetation index (NDVI) determination. Furthermore, the analytical determination of lipid oxidation (MDA level) and antioxidant capacity (FRAP) as well as gene expression studies of the antioxidative enzymes glutathione reductase (GR), glutathione S-transferase (GST) and lipoxygenase (LOX) for essential Zn and nonessential cadmium (Cd) were also carried out in order to clarify toxic symptoms through different Zn investigation approaches. It was possible to identify a metabolic enhancement from 1000 µM; however, stress symptoms from the 2000 µM Zn treatment were noted for both the investigated photosynthetic and oxidative processes. The outcomes of this research contribute to the improvement of Zn mineral-supplementation technology, which is essential for maize growth, and the optimization of agricultural practices.

Effect of Electrostatic Field Assisted Thawing on the Quality of Previously Frozen Beef Striploins

The objective of this study was to evaluate the impact of applying electrostatic field (EF)–assisted thawing on the quality attributes of previously frozen beef striploin. Beef striploins from both sides of 12 USDA Choice carcasses were halved, frozen at −40°C, and thawed under 4 EF voltage treatments: 0 kV (control), 2.5 kV, 5 kV, and 10 kV. After reaching the internal temperature of −1°C, striploins were weighed for yield calculation, swabbed for microbial analysis, fabricated into steaks, and assigned to either 0- or 14-d aging period and retail displayed for 0 or 7 d. Subjective and objective color measurements were taken during the retail display. Upon completion of retail display, Warner-Bratzler shear force (WBSF), cook loss, sarcomere length, troponin-T degradation, muscle fiber spacing, lipid oxidation, antioxidant capacity, pH, and proximate analysis were performed. All EF treatments increased purge loss compared to the control (P<0.05) and did not improve thawing speed, with samples from 10 kV actually taking the longest to thaw (P<0.05). The 2.5 kV and 5 kV samples aged 14 d showed less discoloration than those from 0 kV and 10 kV, and 5 kV samples aged 14 d had higher a* than those from the other treatments (P<0.05). Samples thawed under 10 kV showed a reduction in WBSF compared to the control (P<0.05), but there was no impact of EF on aerobic plate count, sarcomere length, troponin-T degradation,relative fat %, crude protein %, moisture %, purge protein concentration, pH, lipid oxidation, or antioxidant capacity for either the hydrophilic (water soluble) and lipophilic (lipid soluble) portion of the samples (P>0.05). Overall, our study determined that there was no economic benefit to apply EF during thawing regarding yield and purge loss. However, the application of EF may improve tenderness and extend shelf life of beef during retail display.

Analytical Methods for Measuring Lipid Oxidation and Antioxidant Capacity in Food Systems

Analytical Methods for Measuring Lipid Oxidation and Antioxidant Capacity in Food Systems

Embryo thermal manipulation modifies development and hepatic lipid metabolism in post-hatch layer-type chicks.

Incubation temperature is a crucial environmental factor affecting embryonic development and chick quality. Metabolism during the embryonic stage, particularly liver lipid metabolism, is essential for the growth and development of poultry. This study aimed to investigate the effects of embryo thermal manipulation with high (TMH, 39.5 °C, 65% RH, 8h/d) and low (TML, 20 °C, 65% RH, 1h/d) temperatures during 8th to 15th embryonic age on hatching performance and liver lipid metabolism in layer chicks. Additionally, the duration of TM effects was evaluated through a short-term feeding trial. The results indicated that TMH accelerated the hatching process without significantly affecting hatchability and growth performance. In contrast, TML delayed hatching time and significantly reduced hatchability and chick quality. After hatching, TML also increased residual yolk weight and reduced the relative liver weight in relation to body weight and yolk-free body mass. Moreover, lipid droplets in the liver were stained with Oil Red O, and the lipid content in the liver and serum was further detected. TMH had no significant impact on triglyceride (TG) and total-cholesterol (TCHO) content in the liver and serum but upregulated the expression of lipogenesis-related genes ACC, Fas, and Fatp1 compared to the TML group. Conversely, TML significantly reduced liver TG content, enhanced lipoprotein lipase (LPL) activity, and promoted the expression of lipid oxidation-related genes CPT-1, PGC-1α, and PPARα. At 7 d of age, liver LPL activity was significantly increased in the TMH group. However, there were no significant changes in the content of TG and TCHO in the liver and the expression of lipid metabolism-related genes in the TML group. Overall, these results indicate that embryonic TM alters hatching performance and liver lipid metabolism in layer chicks. TML reduces TG content by increasing liver lipid oxidation capacity. However, this effect is not long-lasting, as the influence of TM diminishes as the chicks develop.

Effect of Storage Conditions on Physical Properties, Lipid Oxidation, Isoflavones and Antioxidant Capacity of Flour Prepared from Soy Milk By-Product

During the production of soy milk, a by-product is produced, which is typically treated as a waste material. This by-product has significant levels of dietary fibre, proteins, isoflavones and antioxidant capacity. As such, it has been recommended as an effective functional ingredient when milled to a flour after drying at 100 °C. The shelf-life stability of this dried by-product is relatively unknown when stored under different storage conditions (2 °C, 20 °C and 40 °C) and initial moisture content (9%, 12% and 14%), both packaged and exposed to immediate environments. This study investigated the impact of storage over ten weeks on the physical properties, lipid oxidation, antioxidant capacity and stability of isoflavones of this functional ingredient. The results showed that exposure significantly affected the stability of flour, especially on moisture content, water activity, isoflavone concentration and lipid oxidation. Different initial moisture contents significantly affected the initial and final colour, alongside final moisture and water activity when stored covered at 2 °C and 20 °C. Different storage temperatures were found to affect the moisture content, water activity, lipid oxidation, conversion of isoflavones and antioxidant capacity, with all tested initial moisture contents. Of note, higher conversion rates of malonyl isoflavones to β-glucosides were observed at high temperatures and longer times, while a minimum change in aglycone content occurred. In conclusion, the stability of this flour is least influenced by the initial moisture content but is more affected by high storage temperature and unpackaged conditions.

Consistent changes in muscle metabolism underlie dive performance across multiple lineages of diving ducks.

Diving animals must sustain high activity with limited O2 stores to successfully capture prey. Studies suggest that increasing body O2 stores supports breath-hold diving, but less is known about metabolic specializations that underlie underwater locomotion. We measured maximal activities of 10 key enzymes in locomotory muscles (gastrocnemius and pectoralis) to identify biochemical changes associated with diving in pathways of oxidative and substrate-level phosphorylation and compared them across three groups of ducks-the longest diving sea ducks (eight spp.), the mid-tier diving pochards (three spp.) and the non-diving dabblers (five spp.). Relative to dabblers, both diving groups had increased activities of succinate dehydrogenase and cytochrome c oxidase, and sea ducks further showed increases in citrate synthase (CS) and hydroxyacyl-CoA dehydrogenase (HOAD). Both diving groups had relative decreases in capacity for anaerobic metabolism (lower ratio of lactate dehydrogenase to CS), with sea ducks also showing a greater capacity for oxidative phosphorylation and lipid oxidation (lower ratio of pyruvate kinase to CS, higher ratio of HOAD to hexokinase). These data suggest that the locomotory muscles of diving ducks are specialized for sustaining high rates of aerobic metabolism, emphasizing the importance of body O2 stores for dive performance in these species.

Exploring the potential effect of phospholipase A2 antibody to extend beef shelf-life in a beef liposome model system

The objective of this study was to elucidate the effect of phospholipase A2 (PLA2) and a PLA2 antibody (aPLA2) on phospholipid (PL) hydrolysis in beef and to understand how the altered PL composition may affect lipid oxidation and antioxidant capacity of beef in an in vitro system. Various combinations of PLA2 and aPLA2 were introduced to a beef liposome model system and exposed to a retail display. The PL and free fatty acid (FFA) profiles, antioxidant capacity and lipid oxidation were measured for the liposome system. Key PL classes were reduced and the release of polyunsaturated FFAs was increased with the inclusion of PLA2 in the treatments (P < 0.05). There was no inhibition of PL hydrolysis with the addition of aPLA2. PLA2 showed strong antioxidant capacity in the liposome system (P < 0.01), but lipid oxidation still increased in samples treated with PLA2 throughout the retail display (P < 0.01). Finally, aPLA2 treatments demonstrated potential to decrease lipid oxidation (P < 0.01).

Effects of sodium chloride and sodium tripolyphosphate on the prooxidant properties of hemoglobin in washed turkey muscle system.

This study examined the effects of sodium chloride (NaCl) and sodium tripolyphosphate (STPP) on lipid oxidation induced by oxyhemoglobin (oxyHb) in washed turkey muscle (WTM) model. To explore the reasons for observed effects, the pro-oxidant abilities of Hb derivatives (e.g., metHb, oxyHb, hemin, Fe2+, and Fe3+), pH change, and antioxidation of Hb in the presence of NaCl or STPP were also analyzed. The observed lipid oxidation capacity in WTM followed the order metHb>hemin>oxyHb>Fe2+ > Fe3+. Added Fe2+ accelerated auto-oxidation of oxyHb and oxyHb-mediated lipid oxidation. Hb auto-oxidation to metHb increased as the pH decreased from 6.6 to 5.0. NaCl promoted oxyHb-mediated lipid oxidation due to NaCl causing decreased pH value and increased formation of metHb. STPP inhibited oxyHb-mediated lipid oxidation and weakened the pro-oxidative effect of NaCl. This could be attributed to STPP increasing the pH, inactivating free iron, and inhibiting formation of metHb.

The effects of training, acute exercise and dietary fatty acid composition on muscle lipid oxidative capacity in European starlings.

Migratory birds undergo seasonal changes to muscle biochemistry. Nonetheless, it is unclear to what extent these changes are attributable to the exercise of flight itself versus endogenous changes. Using starlings (Sturnus vulgaris) flying in a wind tunnel, we tested the effects of exercise training, a single bout of flight and dietary lipid composition on pectoralis muscle oxidative enzymes and lipid transporters. Starlings were either unexercised or trained over 2 weeks to fly in a wind tunnel and sampled either immediately following a long flight at the end of this training or after 2 days recovery from this flight. Additionally, they were divided into dietary groups that differed in dietary fatty acid composition (high polyunsaturates versus high monounsaturates) and amount of dietary antioxidant. Trained starlings had elevated (19%) carnitine palmitoyl transferase and elevated (11%) hydroxyacyl-CoA dehydrogenase in pectoralis muscle compared with unexercised controls, but training alone had little effect on lipid transporters. Immediately following a long wind-tunnel flight, starling pectoralis had upregulated lipid transporter mRNA (heart-type fatty acid binding protein, H-FABP, 4.7-fold; fatty acid translocase, 1.9-fold; plasma membrane fatty acid binding protein, 1.6-fold), and upregulated H-FABP protein (68%). Dietary fatty acid composition and the amount of dietary antioxidants had no effect on muscle catabolic enzymes or lipid transporter expression. Our results demonstrate that birds undergo rapid upregulation of catabolic capacity that largely becomes available during flight itself, with minor effects due to training. These effects likely combine with endogenous seasonal changes to create the migratory phenotype observed in the wild.

Resistance to obesity prevents obesity development without increasing spontaneous physical activity and not directly related to greater metabolic and oxidative capacity.

There are evidence that obese-resistant animals are more physically active, due to a higher rate of lipid oxidation. Efficiency in such pathways can favor greater spontaneous physical activity and, consequently, less body fat deposition. The aim of study was characterizing the nutritional profile and spontaneous physical activity in the condition of Resistance to Obesity (OR). Wistar rats were randomized into standard diet (SD; n = 50) and high-fat diet (HFD; n = 50) groups, after obesity induction, were redistributed into Control (C), False-control (FC), Propensity to obesity (OP) and OR, and then spontaneous physical activity was evaluated. Analyzed parameters: body mass (BM), epididymal (EF), retroperitoneal (RF), visceral (VF) and respective summations (∑), adiposity index (AI), nutritional, morphological, biochemical and metabolic parameters and protein quantification. The comparison of the groups was performed by ANOVA one or two factors, with 5% significance adopted. OP and FC presented high final MC values compared to C and OR. OR had lower EF, RF, VF, ∑ and IA compared to OP. OR had similar values to C and higher HDL than FC and OP. In GTT, OR and C presented similar values and both were lower than OP in the 30 minutes. OP promoted higher values than C for glycemic AUC. OR had higher PPARγ content than C and OP, as well as levels similar to C for leptin and insulin. Spontaneous physical activity did not differ between groups. The results were not enough to show that OR animals have greater lipid oxidative capacity, as well as greater spontaneous physical activity.

Metabolic consequences of PGC-1α dysregulation in adult zebrafish muscle.

The peroxisome proliferator-activated receptor γ coactivator 1 α (PGC-1α) is central to the regulation of cellular and mitochondrial energy homeostasis in mammals, but its role in other vertebrates remains unclear. Indeed, previous work suggests extensive structural and functional divergence of PGC-1α in teleosts but this remains to be directly tested. Here, we describe the initial characterization of heterozygous PGC-1α mutant zebrafish lines created by CRISPR-Cas9 disruptions of an evolutionarily conserved regulatory region of the PGC-1α proximal promoter. Using qPCR, we confirmed the disruption of PGC-1α gene expression in striated muscle, leading to a simultaneous fourfold increase in mixed skeletal muscle PGC-1α mRNA levels and an opposite fourfold downregulation in cardiac muscle. In mixed skeletal muscle, most downstream effector genes were largely unaffected yet two mitochondrial lipid transporters, carnitine palmitoyltransferase-1 and -2, were strongly induced. Conversely, PGC-1α depression in cardiac muscle reduced the expression of several transcriptional regulators (estrogen-related receptor α, nuclear respiratory factor 1, and PGC-1β) without altering metabolic gene expression. Using high-resolution respirometry, we determined that white muscle exhibited increased lipid oxidative capacity with little difference in markers of mitochondrial abundance. Finally, using whole animal intermittent respirometry, we show that mutant fish exhibit a twofold higher basal metabolism than their wild-type counterparts. Altogether, this new model confirms a central but complex role for PGC-1α in mediating energy utilization in zebrafish, and we propose its use as a valuable tool to explore the intricate regulatory pathways of energy homeostasis in a popular biomedical model.

Characteristic and effect analysis of protein and peptide in Cantonese cured meat processing

The aim of this work was to explore the physicochemical and structural properties, lipid oxidation and antioxidant capacity of the peptides extracted from Cantonese cured meat and as well as to investigate the effect of drying time on the sarcoplasmic and myofibrillar proteins of Cantonese cured meat. The results suggested that salting out, protein oxidation and heat treatment were closely related to surface hydrophobicity and the secondary structure of peptides was changed by processing. And the peroxide value and the value of tributyltin compounds were different in evaluating the degree of lipid oxidation. Glu and His were the major amino acid. The approximate molecular weights of the sarcoplasmic proteins and myofibrillar proteins ranged from 31 kDa to 50 kDa and 66 kDa, respectively. The results indicated that reducing the levels of protein oxidation and improvement of the antioxidant properties should be of great interest to preserve the nutritional quality of meat products and prolong preservation period.

Contribution of skeletal muscle and serum lipids to muscle contraction induced by neuromuscular electrical stimulation in older individuals.

Intramyocellular lipids (IMCL) stored in droplets in muscle cells and free fatty acids (FFA) from fat cells in the blood are the main substrates of adenosine triphosphate during continuous muscle contractions of relatively lower intensity. Although it is known that the lipid oxidative capacity decreases with aging, the effect of IMCL and FFA on muscle contraction in older individuals remains unclear. The purpose of this study was to investigate the contribution of skeletal muscle lipids and blood lipids as energy sources for muscle contraction in older individuals. Eighteen older individuals (mean age: 70.4 ± 3.5 years) underwent muscle contraction intervention induced by intermittent neuromuscular electrical stimulation (NMES) to the vastus lateralis for 30 min. Fasting blood samples were obtained and proton magnetic resonance spectroscopy (1H‐MRS) was performed before and after NMES, and the parameters (including IMCL and extramyocellular lipid [EMCL]) from 1H‐MRS, along with FFA and adiponectin levels, were analyzed using the blood samples of all participants. Levels of IMCL and EMCL did not change (p > 0.05); however, FFA and adiponectin levels decreased from 1.1 ± 0.5 mEq/L to 0.8 ± 0.2 mEq/L and 12.0 ± 5.3 μg/ml to 11.4 ± 5.0 μg/ml, after NMES (p < 0.05), respectively. These findings indicate that serum lipids, but not skeletal muscle lipids, are the energy substrate utilized during involuntary muscle contraction in older individuals.

Analytical Methods for Lipid Oxidation and Antioxidant Capacity in Food Systems.

Lipid oxidation is the most crucial quality parameter in foods. Many methods were developed to determine the level of oxidation and antioxidant activity. This review compares the methods used to determine lipid oxidation and antioxidant capacity in foods. Lipid oxidation methods developed are based on the direct or indirect measurement of produced primary or secondary oxidation substances. Peroxide values and conjugated diene methods determine the primary oxidative products of lipid oxidation and are commonly used for plant oils and high-fat products. 2-Thiobarbituric acid-reactive substances and chromatographic methods are used to determine the secondary products of oxidation and are suitable for meat and meat-based products. The fluorometric and sensory analyses are indirect methods. The antioxidant capacity of additives is determined indirectly using the lipid oxidation methods mentioned above or directly based on the free-radical scavenging activity of the antioxidant compounds. Each lipid oxidation and antioxidant capacity methods use different approaches, and one method cannot be used for all foods. Therefore, selecting proper methods for specific foods is essential for accurately evaluating lipid oxidation or antioxidant capacity.

Female Mice Are Protected from Metabolic Decline Associated with Lack of Skeletal Muscle HuR.

Simple SummaryMetabolic flexibility describes the ability to adapt to utilization of metabolic fuels such as carbohydrates, lipids, and proteins as they become available. The RNA binding protein HuR controls this flexibility in mouse and human skeletal muscle, but the molecular mechanisms governing this process remain poorly characterized. Additionally, studies from mice indicate that HuR control of metabolic flexibility may be more essential for males than females. This is because males lacking HuR in skeletal muscle develop hallmarks of insulin sensitivity, while females have not been shown to do so. Here we examine this sexual dimorphism in mice lacking HuR in skeletal muscle. Our results reveal that lack of HuR in skeletal muscle drives increased adiposity regardless of sex, but that this increase in adiposity drives the development of insulin resistance in male animals only. Additionally, relative to male mice, the detrimental metabolic phenotype associated with HuR inhibition in skeletal muscle can be corrected by feeding of a diet heavily composed of either lipids or carbohydrates.Male mice lacking HuR in skeletal muscle (HuRm−/−) have been shown to have decreased gastrocnemius lipid oxidation and increased adiposity and insulin resistance. The same consequences have not been documented in female HuRm−/− mice. Here we examine this sexually dimorphic phenotype. HuRm−/− mice have an increased fat mass to lean mass ratio (FM/LM) relative to controls where food intake is similar. Increased body weight for male mice correlates with increased blood glucose during glucose tolerance tests (GTT), suggesting increased fat mass in male HuRm−/− mice as a driver of decreased glucose clearance. However, HuRm−/− female mice show decreased blood glucose levels during GTT relative to controls. HuRm−/− mice display decreased palmitate oxidation in skeletal muscle relative to controls. This difference is more robust for male HuRm−/− mice and more exaggerated for both sexes at high dietary fat. A high-fat diet stimulates expression of Pgc1α in HuRm−/− male skeletal muscle, but not in females. However, the lipid oxidation Pparα pathway remains decreased in HuRm−/− male mice relative to controls regardless of diet. This pathway is only decreased in female HuRm−/− mice fed high fat diet. A decreased capacity for lipid oxidation in skeletal muscle in the absence of HuR may thus be linked to decreased glucose clearance in male but not female mice.

The Influence of High‐Fat Feeding and Exercise Training on Substrate‐Specific Mitochondrial Respiration in Skeletal Muscle of Female Mice

Objective High-fat diet (HFD) induced obesity is associated with excessive lipid supply which drives remodeling of skeletal muscle mitochondria. Exercise stimulates mitochondrial respiration but it is not clear if such adaptations are specific to lipids. Our objective was to determine if HFD-induced obesity would change the sensitivity and capacity for lipid oxidation, and if exercise modifies mitochondrial-specific respiration during HFD. Hypothesis We hypothesized that HFD-induced obesity would increase the sensitivity (intrinsic function) and capacity for lipid respiration while exercise would further augment overall capacity for skeletal muscle mitochondrial substrate oxidation in female mice. Methods We used a 2x2 designof diet and exercise in 10-week old female C57BL/6J mice (n=13-15 per group). Mice consumed diet (either 10% fat or 60% HFD) for 4 weeks then performed 8 weeks of treadmill training for 1 hour per day, 5 days per week or remained sedentary while continuing diet. HFD increased body-weight (P<0.01) by 4 weeks and fat mass measured by dual-energy x-ray absorptiometry at week 12 (P<0.01). We confirmed glucose and insulin intolerance at week 4 of HFD with no change after 8 weeks of treadmill training (P=0.435). Mice rested 24 hours prior to tissue collection. We isolated mitochondria from fresh quadriceps then performed high-resolution respirometry. We measured lipid sensitivity (Km) and capacity (Vmax) using titrations of octanoyl-carnitine (F-linked). Glutamate-succinate were used to determine non-lipid substrate oxidative phosphorylation capacity (N- and S-linked). Primary outcomes were analyzed using 2-way ANOVA (diet x exercise) with Tukey's multiple comparisons (α=0.05). Results The Km for lipid substrates was not different between diet (-7.7%, P=0.66) or exercise (+20.3%, P=0.26) groups. Absolute Vmax was 38% higher in HFD (P=0.018 versus low fat). There was a tendency for 27% higher Vmax after exercise (P=0.074 versus sedentary). For non-lipid substrates, HFD had 23% greater absolute N-linked respiration (P=0.04) and 19% greater S-linked respiration (P=0.02); however, no effects of exercise were detected. Normalizing respiration to mitochondrial protein content eliminated the HFD effects in all substrates. Conclusion We provide evidence that mitochondrial adaptations to HFD, in female mice, are driven primarily by increases in protein abundance rather than intrinsic factors as the normalization of respiration to mitochondrial protein content eliminated HFD effects for increased substrate respiration and there were no differences to lipid sensitivity between diet or exercise. Exercise did not appear to alter the mitochondrial respiratory response to HFD.

Short-Term High-Fat Feeding Does Not Alter Mitochondrial Lipid Respiratory Capacity but Triggers Mitophagy Response in Skeletal Muscle of Mice.

Lipid overload of the mitochondria is linked to the development of insulin resistance in skeletal muscle which may be a contributing factor to the progression of type 2 diabetes during obesity. The targeted degradation of mitochondria through autophagy, termed mitophagy, contributes to the mitochondrial adaptive response to changes in dietary fat. Our previous work demonstrates long-term (2-4 months) consumption of a high-fat diet increases mitochondrial lipid oxidation capacity but does not alter markers of mitophagy in mice. The purpose of this study was to investigate initial stages of mitochondrial respiratory adaptations to high-fat diet and the activation of mitophagy. C57BL/6J mice consumed either a low-fat diet (LFD, 10% fat) or high-fat diet (HFD, 60% fat) for 3 or 7 days. We measured skeletal muscle mitochondrial respiration and protein markers of mitophagy in a mitochondrial-enriched fraction of skeletal muscle. After 3 days of HFD, mice had lower lipid-supported oxidative phosphorylation alongside greater electron leak compared with the LFD group. After 7 days, there were no differences in mitochondrial respiration between diet groups. HFD mice had greater autophagosome formation potential (Beclin-1) and greater activation of mitochondrial autophagy receptors (Bnip3, p62) in isolated mitochondria, but no difference in downstream autophagosome (LC3II) or lysosome (Lamp1) abundance after both 3 and 7 days compared with the LFD groups. In cultured myotubes, palmitate treatment decreased mitochondrial membrane potential and hydrogen peroxide treatment increased accumulation of upstream mitophagy markers. We conclude that several days of high-fat feeding stimulated upstream activation of skeletal muscle mitophagy, potentially through lipid-induced oxidative stress, without downstream changes in respiration.

Low dietary inclusion of nutraceuticals from microalgae improves feed efficiency and modifies intermediary metabolisms in gilthead sea bream (Sparus aurata)

The aim of this work was to evaluate two functional feeds for the gilthead seabream, Sparus aurata, containing low inclusion of two microalgae-based products (LB-GREENboost, LBGb; and LB-GUThealth, LBGh). Fish (12–13 g) were fed for 13 weeks a control diet or one of the four diets supplemented with both products at 0.5% or 1%. LBGb and LBGh did not affect specific growth rate or survival, but increased feed efficiency by decreasing feed intake and enlarging the intestines. LBGb increased hepatosomatic index and reduced cortisol levels in plasma, while both products lowered plasma lactate. Extensive metabolite and metabolic enzyme profiling revealed that microalgae supplementations, especially 1% LBGh: (i) decrease plasma lactate and increase hepatic glycogen, (ii) reduce hepatic gluconeogenesis, (iii) enhance hepatic lipogenic activity and lipid secretion, (iv) led fish to double triglyceride content in muscle and to stimulate its lipid oxidative capacity, and (v) increase the content of monounsaturated fatty acids and the omega-3 alpha-linolenic acid in muscle. This study demonstrates that both microalgae-based products are suited to improve feed efficiency and orchestrate significant changes in the intermediary metabolism in gilthead seabream juveniles.