No effect of beetroot juice supplementation on exercise economy and performance in recreationally active females despite increased torque production.

This study investigated the effects of acute and chronic beetroot juice (BRJ) supplementation on submaximal exercise oxygen uptake (VO2 ), time trial (TT) performance, and contractile properties of the plantar flexors in females. Study 1: Using a double blind, randomized, crossover design, 12 recreationally active females using hormonal contraceptives supplemented acutely (2.5 h) and chronically (8 days) with 280 mL BRJ/d (~26 mmoles nitrate [ NO3- ]) or a NO3- -free placebo (PLA). On days 1 and 8, participants cycled for 10 min at 50% and 70% VO2peak and completed a 4 kJ/kg body mass TT. Plasma [ NO3- ] and nitrite ([NO2- ]) increased significantly following BRJ supplementation versus PLA. There was no effect of BRJ supplementation on VO2 at 50% or 70% VO2peak , or TT performance. Study 2: 12 recreationally active females (n = 7 from Study 1) using hormonal contraceptives participated in a baseline visit and were supplemented acutely (2.5 h) and chronically (8 days) with 280 mL BRJ/d. Maximum voluntary strength (MVC) of the plantar flexors was assessed and a torque-frequency curve performed. BRJ had no effect on MVC, voluntary activation, peak twitch torque, time to peak torque, or half relaxation time. Following both acute (46.6 ± 4.9% of 100 Hz torque) and chronic (47.2 ± 4.4%) supplementation, 10 Hz torque was significantly greater compared to baseline (32.9 ± 2.6%). In summary, BRJ may not be an effective ergogenic aid in recreationally active females as it did not reduce submaximal exercise VO2 or improve aerobic TT performance despite increasing low frequency torque production.

A novel in vitro model for the assessment of postnatal myonuclear accretion.

BACKGROUND: Due to the post-mitotic nature of myonuclei, postnatal myogenesis is essential for skeletal muscle growth, repair, and regeneration. This process is facilitated by satellite cells through proliferation, differentiation, and subsequent fusion with a pre-existing muscle fiber (i.e., myonuclear accretion). Current knowledge of myogenesis is primarily based on the in vitro formation of syncytia from myoblasts, which represents aspects of developmental myogenesis, but may incompletely portray postnatal myogenesis. Therefore, we aimed to develop an in vitro model that better reflects postnatal myogenesis, to study the cell intrinsic and extrinsic processes and signaling involved in the regulation of postnatal myogenesis. METHODS: Proliferating C2C12 myoblasts were trypsinized and co-cultured for 3 days with 5 days differentiated C2C12 myotubes. Postnatal myonuclear accretion was visually assessed by live cell time-lapse imaging and cell tracing by cell labeling with Vybrant® DiD and DiO. Furthermore, a Cre/LoxP-based cell system was developed to semi-quantitatively assess in vitro postnatal myonuclear accretion by the conditional expression of luciferase upon myoblast-myotube fusion. Luciferase activity was assessed luminometrically and corrected for total protein content. RESULTS: Live cell time-lapse imaging, staining-based cell tracing, and recombination-dependent luciferase activity, showed the occurrence of postnatal myonuclear accretion in vitro. Treatment of co-cultures with the myogenic factor IGF-I (p < 0.001) and the cytokines IL-13 (p < 0.05) and IL-4 (p < 0.001) increased postnatal myonuclear accretion, while the myogenic inhibitors cytochalasin D (p < 0.001), myostatin (p < 0.05), and TNFα (p < 0.001) decreased postnatal myonuclear accretion. Furthermore, postnatal myonuclear accretion was increased upon recovery from electrical pulse stimulation-induced fiber damage (p < 0.001) and LY29004-induced atrophy (p < 0.001). Moreover, cell type-specific siRNA-mediated knockdown of myomaker in myoblasts (p < 0.001), but not in myotubes, decreased postnatal myonuclear accretion. CONCLUSIONS: We developed a physiologically relevant, sensitive, high-throughput cell system for semi-quantitative assessment of in vitro postnatal myonuclear accretion, which can be used to mimic physiological myogenesis triggers, and can distinguish the cell type-specific roles of signals and responses in the regulation of postnatal myogenesis. As such, this method is suitable for both basal and translational research on the regulation of postnatal myogenesis, and will improve our understanding of muscle pathologies that result from impaired satellite cell number or function.