Essential Amino Acid-Enriched Diet Alleviates Dexamethasone-Induced Loss of Muscle Mass and Function through Stimulation of Myofibrillar Protein Synthesis and Improves Glucose Metabolism in Mice

Dexamethasone (DEX) induces dysregulation of protein turnover, leading to muscle atrophy and impairment of glucose metabolism. Positive protein balance, i.e., rate of protein synthesis exceeding rate of protein degradation, can be induced by dietary essential amino acids (EAAs). In this study, we investigated the roles of an EAA-enriched diet in the regulation of muscle proteostasis and its impact on glucose metabolism in the DEX-induced muscle atrophy model. Mice were fed normal chow or EAA-enriched chow and were given daily injections of DEX over 10 days. We determined muscle mass and functions using treadmill running and ladder climbing exercises, protein kinetics using the D2O labeling method, molecular signaling using immunoblot analysis, and glucose metabolism using a U-13C6 glucose tracer during oral glucose tolerance test (OGTT). The EAA-enriched diet increased muscle mass, strength, and myofibrillar protein synthesis rate, concurrent with improved glucose metabolism (i.e., reduced plasma insulin concentrations and increased insulin sensitivity) during the OGTT. The U-13C6 glucose tracing revealed that the EAA-enriched diet increased glucose uptake and subsequent glycolytic flux. In sum, our results demonstrate a vital role for the EAA-enriched diet in alleviating the DEX-induced muscle atrophy through stimulation of myofibrillar proteins synthesis, which was associated with improved glucose metabolism.

Comparison of the Cardiovascular Effects of Extreme Psychological and Physical Stress Tests in Male Soccer Players

Background: The purpose of our study was to compare the physiological effects of extreme physical and psychological stress tests in male soccer players, since these two types of stress apply to athletes with high performance requirements. Methods: A total of 63 healthy male soccer players participated in this study, all of whom underwent both of the tests. A physical stress test was carried out in an exercise physiology laboratory, where subjects completed an incremental treadmill running test to full exhaustion, and a psychological test was performed in a military tactical room, where subjects met a street offence situation. Heart rate variability (HRV) and blood pressure (BP) were recorded directly before, immediately after, and 30 min after the stress tests. Results: The majority of HRV indices changed significantly in both stress protocols. Inverse, significant changes (positive for the physical test, negative for the psychological test, p < 0.001) were found when comparing the alterations of HRV indices between the tests. Significant differences were found in the changes in systolic (p = 0.003) and diastolic (p < 0.001) BP between the test protocols, and also between the baseline and post-test measurements (p < 0.001). Conclusion: Both HRV and BP are sensitive physiological parameters to measure the impact of extreme physical and/or psychological stress

Receptor interacting protein kinase-3 promotes both myopathy and cardiomyopathy in dystrophin-deficient mice

Background Duchenne muscular dystrophy (DMD) is a muscle degenerative disorder that is caused by the absence of dystrophin. From early childhood, multiple rounds of myofibre necrosis and regeneration lead to fibrosis and fat deposition, irreversibly disturbing skeletal muscle function and impairing locomotion. Cell necrosis also affects respiratory muscles and cardiomyocytes, ultimately responsible for the death of DMD boys by respiratory or heart failure. Necroptosis is a genetically programmed form of necrosis requiring the receptor-interacting serine/threonine-protein kinase (RIPK)3 and is a promising new therapeutic target for multiple degenerative disorders. We previously demonstrated that necroptosis mediates hindlimb myofibre degeneration in distinct muscular dystrophies, including in DMD. However, this pathway was recently found to be required for myogenesis. Its prevention might therefore lead to detrimental side effects on muscle repair. Whether necroptosis also participates in the pathogenesis of respiratory and cardiac muscle dysfunction, and whether its long-term inhibition would ultimately be beneficial or detrimental to mdx mice are addressed here. Methods Herein, we examined the effects of RIPK3 depletion on an advanced stage of pathogenesis in mdx mice. Dystrophic mice aged 12 to 18 months were submitted to forced treadmill running to assess their locomotor function. mdx cardiomyopathy was also examined by echocardiography in 40-week-old mice. Limb skeletal muscles, diaphragm and heart were analyzed by histology and molecular biology to compare the phenotype of mdxRipk3+/+ mdxRipk3-/- mice. Results In 18-month-old mdxRipk3-/- mice, we found no sign of muscle regeneration defect compared to mdxRipk3+/+ littermates. mdxRipk3-/- mice had decreased fibrosis in limb muscles, without evidence of muscle atrophy. The size of diaphragm myofibres was slightly reduced and affected by less variability than mdx littermates. Fibrosis was also reduced in the diaphragm of RIPK3-deficient mdx mice. Notably, heart hypertrophy and left ventricle fibrosis were reduced in mdxRipk3-/- mice, and using echocardiography, we found a significant decrease of markers of cardiomyopathy by such as a reduction of the relative wall thickness and left ventricle mass. Conclusions Our data suggest that necroptosis is involved together in the pathogenic phenotype of locomotor, respiratory, and cardiac muscles in dystrophin-deficient mice. The long-term genetic ablation of RIPK3 does not generate evidence of sarcopenia or muscle impairment in mdx mice. Our data suggest that necroptosis may represent a new therapeutic target susceptible to improving the phenotype of myopathy and cardiomyopathy.

Using Ground Reaction Forces To Predict Fatigue In Treadmill Running: A Machine Learning Approach

The analysis of running patterns, especially those associated with fatigue, can help specialists in designing more efficient workouts and preventing injuries in high-performance sports. However, classifying running patterns is not trivial for humans. An interesting alternative is to use Machine Learning methods, such as Artificial Neural Networks (ANNs), to classify running patterns. In this work, ground reaction forces are measured by sensors coupled to the base of a low-cost open-source treadmill. ANNs are used to classify the force signals and to indicate the occurrence of fatigue. Different features, extracted from the force signals, are proposed and investigated. A Genetic Algorithm (GA) is used to select the best features. The experimental results indicate that the ANN is able to classify the running patterns with good accuracy. In addition, some features selected by the GA provide important information regarding the identification of fatigue in treadmill running.

Long-term light and moderate exercise intervention similarly prevent both hippocampal and glycemic dysfunction in pre-symptomatic type 2 diabetic rats

A pre-diabetic population has an increased risk of cognitive decline as well as type 2 diabetes mellitus (T2DM). The present study investigated whether the progression of memory dysfunction and dysregulated brain glycogen metabolism is prevented with four months of exercise intervention from the pre-symptomatic stage in T2DM rat model. Memory function and biochemical and molecular profiles were assessed in the pre-symptomatic stage of OLETF rats, a T2DM model, with LETO rats as genetic control. These rats were subjected to light- or moderate-intensity treadmill running for four months with repetition of the same experiments. Significant hippocampal-dependent memory dysfunction was observed in the pre-symptomatic stage of OLETF rats, accompanied by downregulated levels of hippocampal monocarboxylate transporter 2 (MCT2), a neuronal lactate-transporter, without alteration in hippocampal glycogen levels. Four months of light or moderate exercise from the pre-symptomatic stage of T2DM normalized glycemic parameters and also hippocampal molecular normalization through MCT2, glycogen, and brain-derived neurotrophic factor (BDNF) levels with the improvement of memory dysfunction in OLETF rats. A four-month exercise regimen from the pre-symptomatic stage of T2DM at light and moderate intensities contributed to the prevention of the development of T2DM and the progression of cognitive decline with hippocampal lactate-transport and BDNF improvement.

An open-source, lockable mouse wheel for the accessible implementation of time- and distance-limited elective exercise

Current methods of small animal exercise involve either voluntary (wheel running) or forced (treadmill running) protocols. Although commonly used, each have several drawbacks which cause hesitancy to adopt these methods. While mice will instinctively run on a wheel, the distance and time spent running can vary widely. Forced exercise, while controllable, puts animals in stressful environments in which they are confined and often shocked for “encouragement.” Additionally, both methods require expensive equipment and software, which limit these experiments to well-funded laboratories. To counter these issues, we developed a non-invasive mouse running device aimed to reduce handler-induced stress, provide time- and distance-based stopping conditions, and enable investigators with limited resources to easily produce and use the device. The Lockable Open-Source Training-Wheel (LOST-Wheel) was designed to be 3D printed on any standard entry-level printer and assembled using a few common tools for around 20 USD. It features an on-board screen and is capable of tracking distances, running time, and velocities of mice. The LOST-Wheel overcomes the largest drawback to voluntary exercise, which is the inability to control when and how long mice run, using a servo driven mechanism that locks and unlocks the running surface according to the protocol of the investigator. While the LOST-Wheel can be used without a computer connection, we designed an accompanying application to provide scientists with additional analyses. The LOST-Wheel Logger, an R-based application, displays milestones and plots on a user-friendly dashboard. Using the LOST-Wheel, we implemented a timed running experiment that showed distance-dependent decreases in serum myostatin as well as IL-6 gene upregulation in muscle. To make this device accessible, we are releasing the designs, application, and manual in an open-source format. The implementation of the LOST-Wheel and future iterations will improve upon existing murine exercise equipment and research.

Symptom Locus and Symptom Origin Incongruity in Runner’s Dystonia – Case Study of an Elite Female Runner

Objectives: Runner’s dystonia is a task-specific dystonia that occurs in the lower limbs and trunk, with diverse symptomatology. We aimed to identify the origin of a dystonic movement abnormality using combined three-dimensional kinematic analysis and electromyographic (EMG) assessment during treadmill running.Participant: A 20-year-old female runner who complained of right-foot collision with the left-leg during right-leg swing-phase, which mimicked right-ankle focal dystonia.Results: Kinematic and EMG assessment of her running motion was performed, which showed a significant drop of the left pelvis during right-leg stance-phase, and a simultaneous increase of right hip adductor muscle activity. This resulted in a pronounced adduction of the entire right lower limb with respect to the pelvis segment. Trajectories of right foot were seen to encroach upon left-leg area.Discussion: These findings suggested that the symptom of this runner was most likely a form of segmental dystonia originating from an impaired control of hip and pelvis, rather than a distal focal ankle dystonia.Conclusion: We conclude that, for individualized symptom assessment, deconstructing the symptom origin from its secondary compensatory movement is crucial for characterizing dystonia. Kinematic and EMG evaluation will therefore be a prerequisite to distinguish symptom origin from secondary compensatory movement.

The Effect of High-Fat Diet and Exercise Intervention on the TNF-α Level in Rat Spleen

High-fat diet (HFD) consumption can trigger chronic inflammation in some tissues. However, it remains unclear if HFD induces chronic inflammation in the spleen. This investigation aims to address the effect of HFD consumption and exercise intervention on the level of tumor necrosis factor alpha (TNF-α) in the spleen. Rats were subjected to HFD feeding and/or moderate-intensity treadmill running. The TNF-α levels in plasma and spleen were detected by ELISA. The mass and total cell numbers of the spleen were measured. In addition, the expression of TNF-α and its relevant gene mRNAs in macrophages from the spleen were analyzed by qRT-PCR. We found that HFD consumption did not significantly affect the mass and total cell numbers of the spleen. However, HFD consumption significantly increased splenic TNF-α level, the expression of TNF-α, toll-like receptor 4, and nuclear factor κB p65 mRNAs. In contrast, the expression of nicotinic acetylcholine receptor alpha 7 subunit (α7nAChR) mRNA in macrophages was downregulated. Additionally, exercise abolished the increase in splenic TNF-α level as well as the abnormal expression of TNF-α and related gene mRNAs in macrophages in HFD-fed rats. In conclusion, our results reveal that HFD consumption increases TNF-α level in the spleen, which is along with upregulation of the expression of TLR4 and NF-κB mRNAs as well as downregulation of the expression of α7nAChR mRNA in splenic macrophages in rats. Exercise abolished detrimental effects of HFD on TNF-α level in the spleen and prevented abnormal expression of these genes in the macrophages from rat spleen.

Enhanced Oxygen Utilization Efficiency With Concomitant Activation of AMPK-TBC1D1 Signaling Nexus in Cyclophilin-D Conditional Knockout Mice

We have previously reported in HEK 293 T cells and in constitutive cyclophilin-D (Cyp-D) knockout (KO) mice that Cyp-D ablation downregulates oxygen consumption (VO2) and triggers an adaptive response that manifest in higher exercise endurance with less VO2. This adaptive response involves a metabolic switch toward preferential utilization of glucose via AMPK-TBC1D1 signaling nexus. We now investigated whether a similar response could be triggered in mice after acute ablation of Cyp-D using tamoxifen-induced ROSA26-Cre-mediated (i.e., conditional KO, CKO) by subjecting them to treadmill exercise involving five running sessions. At their first treadmill running session, CKO mice and controls had comparable VO2 (208.4 ± 17.9 vs. 209.1 ± 16.8 ml/kg min−1), VCO2 (183.6 ± 17.2 vs. 184.8 ± 16.9 ml/kg min−1), and RER (0.88 ± 0.043 vs. 0.88 ± 0.042). With subsequent sessions, CKO mice displayed more prominent reduction in VO2 (genotype &amp; session interaction p = 0.000) with less prominent reduction in VCO2 resulting in significantly increased RER (genotype and session interaction p = 0.013). The increase in RER was consistent with preferential utilization of glucose as respiratory substrate (4.6 ± 0.8 vs. 4.0 ± 0.9 mg/min, p = 0.003). CKO mice also performed a significantly higher treadmill work for given VO2 expressed as a power/VO2 ratio (7.4 ± 0.2 × 10−3 vs. 6.7 ± 0.2 10−3 ratio, p = 0.025). Analysis of CKO skeletal muscle tissue after completion of five treadmill running sessions showed enhanced AMPK activation (0.669 ± 0.06 vs. 0.409 ± 0.11 pAMPK/β-tubulin ratio, p = 0.005) and TBC1D1 inactivation (0.877 ± 0.16 vs. 0.565 ± 0.09 pTBC1D1/β-tubulin ratio, p &lt; 0.05) accompanied by increased glucose transporter-4 levels consistent with activation of the AMPK-TBC1D1 signaling nexus enabling increased glucose utilization. Taken together, our study demonstrates that like constitutive Cyp-D ablation, acute Cyp-D ablation also induces a state of increased O2 utilization efficiency, paving the way for exploring the use of pharmacological approach to elicit the same response, which could be beneficial under O2 limiting conditions.