Acute Effect of Popular High-Intensity Functional Training Exercise on Physiologic Markers of Growth.

ABSTRACT: Kliszczewicz, B, Markert, CD, Bechke, E, Williamson, C, Clemons, KN, Snarr, RL, and McKenzie, MJ. Acute effect of popular high-intensity functional training exercise on physiologic markers of growth. J Strength Cond Res 35(6): 1677-1684, 2021-Constantly evolving, high-intensity functional training (HIFT) exercise consists of various modalities, orders, weights, and repetition schemes. High-intensity functional training has gained popularity among the general population, but lacks empirical evidence regarding acute adaptive responses. The purpose of this study was to describe the acute effects of 2 representative bouts of HIFT on physiologic markers of growth. For convenience, the bouts are designated "short" (<5 minutes) and "long" (∼15 minutes), although duration was not the only difference between bouts. Ten apparently healthy men (28.1 ± 5 years) performed 2 HIFT bouts in a randomized crossover design. Blood was collected at 5 time points (Pre, Post, 1, 3, and 6 hours) to examine growth hormone (GH), insulin-like growth factor (IGF-1), insulin-like growth factor binding proteins 1 and 2 (IGFBP-1 and IGFBP-2), and vascular endothelial growth factor (VEGF). Blood lactate concentration ([La]) was analyzed at the Pre and Post time points. A repeated-measures analysis of variance (ANOVA) revealed no trial differences among the markers (IGF-1, IGFBP-1, IGFBP-2, and VEGF) except GH at Post, where the long bout produced a greater effect (p = 0.005). Mean GH levels (pg·ml-1) in the short bout increased from 68.4 to 106.5, and in the long bout, mean GH levels increased from 38.5 to 286.4. The repeated-measures ANOVA revealed a main time effect in GH (p = 0.037), while a post hoc t-test demonstrated elevated GH at 1 hour (p = 0.018) when compared with Pre. No time-dependent change (p > 0.05) was observed in IGF-1, IGFBP-1, IGFBP-2, or VEGF. Mean blood [lactate] increased more than tenfold in both bouts. The findings of this descriptive study suggest that, other than GH, there are no acute differences in markers of skeletal muscle or vascular growth between these 2 specific HIFT bouts.

Regenerative medicine: a vehicle to infuse laboratory-bench modules into an exercise physiology curriculum.

Regenerative medicine is a novel discipline that both excites undergraduates and may be used as a vehicle to expose students to scientific concepts and opportunities. The goal of this article is to describe the implementation of a National Science Foundation-funded Targeted Infusion Project in which underrepresented minority undergraduates are exposed to laboratory-bench skills and summer research opportunities that they may not have encountered otherwise. A 3-wk infusion of laboratory-bench and data presentation skills, in the context of a regenerative medicine/bioengineering project, aimed to engage students and expose them to opportunities as summer researchers and teaching assistants. The infusion aimed to assess the extent to which students improved 1) attitudes toward laboratory-bench-based techniques, using attitudes toward science as a proxy; 2) perceptions of scientific inquiry; 3) intentions to engage in undergraduate research; and 4) intentions to persist in science, technology, engineering, and mathematics (STEM)-related fields. Results indicate that the 3-wk infusion had no effect on science attitudes, but transcribed responses to structured interviews administered after the summer research experience indicated that students who completed summer research projects had positive experiences. Differences in intentions to engage in research were detected between groups of students in different STEM majors, in addition to differences in intentions to pursue a career in science. We describe the implementation of the infusion and briefly discuss quantitative outcomes. We conclude that infusion of laboratory-bench modules in the context of a regenerative medicine/bioengineering project may play a small but important role in increasing (minority) participation and persistence in the STEM pipeline.

Dystrophin-deficient cardiomyocytes derived from human urine: new biologic reagents for drug discovery.

The ability to extract somatic cells from a patient and reprogram them to pluripotency opens up new possibilities for personalized medicine. Induced pluripotent stem cells (iPSCs) have been employed to generate beating cardiomyocytes from a patient's skin or blood cells. Here, iPSC methods were used to generate cardiomyocytes starting from the urine of a patient with Duchenne muscular dystrophy (DMD). Urine was chosen as a starting material because it contains adult stem cells called urine-derived stem cells (USCs). USCs express the canonical reprogramming factors c-myc and klf4, and possess high telomerase activity. Pluripotency of urine-derived iPSC clones was confirmed by immunocytochemistry, RT-PCR and teratoma formation. Urine-derived iPSC clones generated from healthy volunteers and a DMD patient were differentiated into beating cardiomyocytes using a series of small molecules in monolayer culture. Results indicate that cardiomyocytes retain the DMD patient's dystrophin mutation. Physiological assays suggest that dystrophin-deficient cardiomyocytes possess phenotypic differences from normal cardiomyocytes. These results demonstrate the feasibility of generating cardiomyocytes from a urine sample and that urine-derived cardiomyocytes retain characteristic features that might be further exploited for mechanistic studies and drug discovery.

Characterizing the micro-scale elastic modulus of hydrogels for use in regenerative medicine.

Our objective was to characterize the elasticity of hydrogel formulations intended to mimic physical properties that cells and tissues experience in vivo. Using atomic force microscopy (AFM), we tested a variety of concentrations in a variety of biomaterials, including agarose, alginate, the collagens, fibrin, hyaluronic acid, kerateine, laminin, Matrigel, polyacrylamide, polyethylene glycol diacrylate (PEGDA) and silicone elastomer (polydimethylsiloxane). Manipulations of the concentration of biomaterials were detectable in AFM measurements of elasticity (Young's modulus, E), and E tended to increase with increased concentration. Depending on the biomaterials chosen, and their concentrations, generation of tunable biocompatible hydrogels in the physiologic range is possible.

Exercise and Duchenne muscular dystrophy: toward evidence-based exercise prescription.

To develop a rational framework for answering questions about the role of exercise in Duchenne muscular dystrophy (DMD), we focused on five pathophysiological mechanisms and offer brief hypotheses regarding how exercise may beneficially modulate pertinent cellular and molecular pathways. We aimed to provide an integrative overview of mechanisms of DMD pathology that may improve or worsen as a result of exercise. We also sought to stimulate discussion of what outcomes/dependent variables most appropriately measure these mechanisms, with the purpose of defining criteria for well-designed, controlled studies of exercise in DMD. The five mechanisms include pathways that are both intrinsic and extrinsic to the diseased muscle cells.

A single-dose resveratrol treatment in a mouse model of amyotrophic lateral sclerosis.

The underlying causes of denervation of the neuromuscular junction and eventual motor neuron death in amyotrophic lateral sclerosis (ALS) have not been resolved. The superoxide dismutase 1 (SOD1)(G93A) mutant mouse is a frequently used animal model of ALS. We hypothesized that resveratrol (RSV), a polyphenolic molecule that enhances mammalian NAD(+)-dependent SIRT1 deacetylases and may increase life span, would improve motor function and survival in the SOD1 mouse model via modulation of p53 acetylation. Data were collected for mean survival times, neuromuscular performance on the ROTOR-ROD™ (San Diego Instruments, San Diego, CA, USA), body weight, and p53 acetylation. Mean survival times were not statistically different (P=.23) between control and experimental (RSV-fed) groups (mean +/- SD, control [n=11] 138 +/- 6 days vs. experimental [n=10] 135 +/- 8 days). Performance was not significantly different between groups at time points corresponding to 50%, 80%, and 90% mean life span (P=.46), nor did RSV treatment attenuate body weight loss. Thus although manipulation of SIRT1 deacetylase activity has effects at the protein level in healthy aging organisms, we conclude that RSV treatment does not lead to functional improvement or increased longevity in a mouse model of ALS. We speculate that RSV-mediated modulation of p53 acetylation is either incapable of increasing or insufficient to increase motor performance and longevity in this model of ALS.

Eccentric contractions induce rapid isometric torque drop in dystrophin-deficient dogs.

We tested the hypothesis that eccentric contractions (ECCs) rapidly induce greater-than-normal isometric torque drop in dystrophin-deficient golden retriever muscular dystrophy (GRMD) muscles. ECCs were imposed by forcibly stretching activated muscles. The results indicate that isometric torque drop was greater in GRMD versus controls (P < 0.0001). Our findings support the hypothesis that ECCs induce greater-than-normal isometric torque drop in GRMD muscles. The magnitude of ECC-induced isometric torque loss may be an ideal clinical endpoint in the GRMD model.

Immunofluorescence microscopy for imaging of nuclear p63 in human primary keratinocytes: a comparison of antibodies and fixation methods.

The ability of an experimental treatment to induce primitive, undifferentiated stem cells towards an epidermal fate may be tested by comparing the treated stem cells with a positive control, such as primary keratinocytes. In an effort to perfect methods used for this comparison, we tested two commercially available antibodies and three fixation methods to determine which antibody/fixation interaction produced the best immunofluorescent images of the nuclear localization of p63, a canonical marker of epidermal fate, in keratinocytes. Here, we report the methods used, and the experimental outcome.

Kinematics of gait in golden retriever muscular dystrophy.

The goal of this study was to quantify the two-dimensional kinematics of pathologic gait during over-ground walking at a self-selected speed at the stifle (knee) and hock (ankle) joints in six Golden Retriever Muscular Dystrophy (GRMD) dogs and six carrier littermates (controls). We found that GRMD dogs walked significantly slower than controls (p<0.01). At the stifle joint, both groups displayed similar ROM (range of motion), but compared to controls, GRMD dogs walked with the stifle joint relatively more extended. At the hock joint, GRMD dogs displayed less ROM (range of motion) and walked with the joint relatively less flexed compared to controls. We controlled for gait speed in all analyses, so the differences we observed in joint kinematics between groups cannot be attributed solely to the slower walking speed of the GRMD dogs. This is the first kinematic study of gait in the GRMD dog, an important step in using this model in pre-clinical trials.

Mesenchymal stem cells: emerging therapy for Duchenne muscular dystrophy.

Multipotent cells that can give rise to bone, cartilage, fat, connective tissue, and skeletal and cardiac muscle are termed mesenchymal stem cells. These cells were first identified in the bone marrow, distinct from blood-forming stem cells. Based on the embryologic derivation, availability, and various pro-regenerative characteristics, research exploring their use in cell therapy shows great promise for patients with degenerative muscle diseases and a number of other conditions. In this review, the authors explore the potential for mesenchymal stem cell therapy in the emerging field of regenerative medicine with a focus on treatment for Duchenne muscular dystrophy.

TCAP knockdown by RNA interference inhibits myoblast differentiation in cultured skeletal muscle cells.

Null mutation of titin-cap (TCAP) causes limb-girdle muscular dystrophy type 2G (LGMD2G). LGMD2G patients develop muscle atrophy, and lose the ability to walk by their third decade. Previous findings suggest that TCAP regulates myostatin, a key regulator of muscle growth. We tested the hypothesis that TCAP knockdown with RNA interference will lead to differential expression of genes involved in muscle proliferation and differentiation, impairing muscle cell growth. mRNA from cultured cells treated with TCAP siRNA duplex constructs was analyzed using Northern blots and real-time RT-PCR. siRNA treatment decreased TCAP mRNA expression in differentiating muscle cells. Significant (p<0.05) decreases in mRNA were observed for myogenic regulatory factors. siRNA treatment also prevented development of the normal phenotype of muscle cells. Our findings suggest that TCAP knockdown with RNA interference alters normal muscle cell differentiation.

Nonthermal ultrasound and exercise in skeletal muscle regeneration.

OBJECTIVE: To determine whether continuous nonthermal therapeutic ultrasound (US) and low-intensity exercise (Ex) influence skeletal muscle regeneration after a standardized contusion injury in an animal model. DESIGN: Randomized controlled trial with blinded comparisons in a 2 x 2 factorial (US by Ex) design. SETTING: Animal care facility and exercise physiology biochemistry laboratory. ANIMALS: Twenty male Wistar rats (age, 8 mo) received a reproducible bilateral contusion injury to the gastrocnemius muscles. Ten gastrocnemius muscles from 5 noninjured, nontreated rats provided baseline control data. INTERVENTIONS: US (continuous duty cycle, 3 MHz; intensity, 0.1 W/cm2 ; transducer, 1cm2 ; duration, 5 min/d; duty cycle, 100%) and exercise (20 min/d of low-intensity treadmill walking at 14 m/min). Gastrocnemius muscles from injured rats received exercise treatment alone (Ex + NoUS), exercise and US treatment (Ex + US), US treatment alone (NoEx + US), and no treatment (NoEx + NoUS). MAIN OUTCOME MEASURES: Ninety-six-hour postinjury muscle mass, contractile protein concentration, fiber cross-sectional area, number of nuclei per fiber, and myonuclear density. RESULTS: Myonuclei per fiber were statistically greater in injured than in noninjured gastrocnemius muscle (P < .05). There were no statistical differences (P > .01) among the 4 injured treatment groups for any of the outcome measures chosen as biomarkers of skeletal muscle regeneration. CONCLUSIONS: There is no evidence that the specific continuous US and Ex protocols investigated enhanced skeletal muscle regeneration after contusion injury.

Age-related changes in metabolic properties of equine skeletal muscle associated with muscle plasticity.

The purpose of the present study was to determine the age-related changes in myosin heavy chain (MHC) composition and muscle oxidative and glycolytic capacity in 18 horses ranging in age from two to 30 years. Muscle samples were collected by excisional biopsy of the semimebranosus muscle. MHC expression and the key enzymatic activities were measured. There was no significant correlation between horse age and the proportions of type-IIA and type-IIX MHC isoforms. The percentage of type-I MHC isoforms decreased with advancing age. Muscle citrate synthase activity decreased, whereas lactate dehydrogenase activity increased with increasing age. Muscle 3-OH acyl CoA dehydrogenase activity did not change with ageing. The results suggest that, similar to humans, the oxidative capacity of equine skeletal muscle decreases with age. The age-related changes in muscle metabolic properties appear to be consistent with an age-related transition in MHC isoforms of equine skeletal muscle that shifts toward more glycolytic isoforms with age.

Exercise training increases oxidative capacity and attenuates exercise-induced ultrastructural damage in skeletal muscle of aged horses.

Exercise training improves functional capacity in aged individuals. Whether such training reduces the severity of exercise-induced muscle damage is unknown. The purpose of the present study was to determine the effect of 10 wk of treadmill exercise training on skeletal muscle oxidative capacity and exercise-induced ultrastructural damage in six aged female Quarter horses (>23 yr of age). The magnitude of ultrastructural muscle damage induced by an incremental exercise test before and after training was determined by electron microscopic examination of samples of triceps, semimembranosus, and masseter (control) muscles. Maximal aerobic capacity increased 22% after 10 wk of exercise training. The percentage of type IIa myosin heavy chain increased in semimembranosus muscle, whereas the percentage of type IIx myosin heavy chain decreased in triceps muscle. After training, triceps muscle showed significant increases in activities of both citrate synthase and 3-hydroxyacyl-CoA-dehydrogenase. Attenuation of exercise-induced ultrastructural muscle damage occurred in the semimembranosus muscle at both the same absolute and the same relative workloads after the 10-wk conditioning period. We conclude that aged horses adapt readily to intense aerobic exercise training with improvements in endurance, whole body aerobic capacity, and muscle oxidative capacity, and heightened resistance to exercise-induced ultrastructural muscle cell damage. However, adaptations may be muscle-group specific.