Physiologia, Volume 1, Issue 1 (December 2021) – 5 articles

Muscle protein synthesis and proteolysis are tightly coupled processes. Given that muscle growth is promoted by increases in net protein balance, it stands to reason that bolstering protein synthesis through amino acids while reducing or inhibiting proteolysis could be a synergistic strategy in enhancing anabolism. However, there is contradictory evidence suggesting that the proper functioning of proteolytic systems in muscle is required for homeostasis. To add clarity to this issue, we sought to determine if inhibiting different proteolytic systems in C2C12 myotubes in conjunction with acute and chronic leucine treatments affected markers of anabolism. In Experiment 1, myotubes underwent 1-h, 6-h, and 24-h treatments with serum and leucine-free DMEM containing the following compounds (n = 6 wells per treatment): (i) DMSO vehicle (CTL), (ii) 2 mM leucine + vehicle (Leu-only), (iii) 2 mM leucine + 40 μM MG132 (20S proteasome inhibitor) (Leu + MG132), (iv) 2 mM leucine + 50 μM calpeptin (calpain inhibitor) (Leu + CALP), and (v) 2 mM leucine + 1 μM 3-methyladenine (autophagy inhibitor) (Leu + 3MA). Protein synthesis levels significantly increased (p < 0.05) in the Leu-only and Leu + 3MA 6-h treatments compared to CTL, and levels were significantly lower in Leu + MG132 and Leu + CALP versus Leu-only and CTL. With 24-h treatments, total protein yield was significantly lower in Leu + MG132 cells versus other treatments. Additionally, the intracellular essential amino acid (EAA) pool was significantly greater in 24-h Leu + MG132 treatments versus other treatments. In a follow-up experiment, myotubes were treated for 48 h with CTL, Leu-only, and Leu + MG132 for morphological assessments. Results indicated Leu + MG132 yielded significantly smaller myotubes compared to CTL and Leu-only. Our data are limited in scope due to the utilization of select proteolysis inhibitors. However, this is the first evidence to suggest proteasome and calpain inhibition with MG132 and CALP, respectively, abrogate leucine-induced protein synthesis in myotubes. Additionally, longer-term Leu + MG132 treatments translated to an atrophy phenotype. Whether or not proteasome inhibition in vivo reduces leucine- or EAA-induced anabolism remains to be determined. Full article

We aimed to analyze the impact of a single CrossFit^® session “workout of the day” (WOD) on plasma redox. Ten CrossFit^®-experienced subjects volunteered to participate. Oxygen uptake (VO₂) during WOD and treadmill running (TR), performed at the same VO₂ and time as WOD, were continuously monitored. Venous blood samples were collected before (baseline—BL) and after both exercises, for lactate concentration, total antioxidant capacity, thiol content, and DNA damage measurements. Total antioxidant capacity decreased after both exercises (WOD and TR) vs. BL, with no differences between exercises. Thiol content increased after WOD; however, no differences between exercises were observed. DNA damage increased after both WOD and TR, although more exuberantly after WOD than TR. Much higher lactate levels were detected in WOD compared to TR. Our findings suggest that WOD induces an increased condition of oxidative injury and affects total antioxidant capacity in experienced CrossFit^® performers. Full article

Caffeine and nitrates have both been reported to enhance performance in power efforts; however, it is not clear which supplement is most effective. The aim of this study was to compare the effects of caffeine and nitrates on the performance of semi-professional soccer players during different fitness tests. Ten male soccer players in a randomized crossover design were assigned to receive caffeine (5 mg/kg body mass) (CG), nitrate ((250 mL/150 mg of NO3⁻) (NG), or a placebo (PG) on three different occasions. In each treatment, the participants performed the following tests: 10 m and 30 m sprints, the Illinois agility test, a countermovement jump test, a squat jump test, and a repeated sprint test (6 × 40 m). Caffeine boosted performance in jumps (CMJ: CGvsPG, p = 0.018; SJ: CGvsPG, p = 0.045 and CGvsNG, p = 0.001) and limited the decrease in performance in the RSA test (CGvsPG, p = 0.012). Nitrates limited the decrease in performance in the RSA test (NGvsPG, p = 0.035). In conclusion, the two supplements limited the decrease in performance in the test of repeated sprints, with caffeine showing a greater effect. Among the other tests, only caffeine improved performance, and only in the jumps. Thus, we can conclude that supplementation with caffeine 1 h before these kinds of activities at a dosage of 5 mg/kg of body weight can enhance performance. Full article