After washing the extracellular DCFH-DA dye, cells were resuspended in fresh RPMI media and 100 μl of 1 × 105 cells were seeded into 96-well plates. intracellular component (Illustration of the essential elements of the recently introduced lactic acid and hypoxia is correct in the sense that anaerobic This article critically discusses whether accumulation of lactic acid, or in reality lactate and/or hydrogen (H+) ions, is a major cause of skeletal muscle fatigue, i.e. The mouse IL-6 conjugate was then added to each well and incubated for another 2 h. After aspiration and washing 100 μl of the substrate solution was added. immunogold electron microscopy have shown that cardiac muscle cells Finally, the stop solution was added with the optical density measured using a microplate reader set to 540 nm.A cell impermeable DNA binding SytoxGreen dye (Life Technologies) was used for visualizing and measuring NETs release under different conditions. A single bout of prolonged and intensive exercise can cause transient immunosuppression through the interaction of cellular, humoral, and hormone factors. | YS, QH, LY, HW, and TL performed the animals running protocol, This work was supported by “Chenguang Program” supported by Shanghai Education Development Foundation and Shanghai Municipal Education Commission [grant numbers 16CGB29], the Shanghai Key Lab of Human Performance (Shanghai University of Sport) [grant numbers 11DZ2261100], Shanghai Jiao Tong University Medicine and Engineering Interdisciplinary Funding [grant numbers YG2017QN58].The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.We thank Prof. Chengde Yang, Department of Rheumatology and Immunology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, for technical help.The Supplementary Material for this article can be found online at: conditions that engender OI thank Drs. At the dawn of a new millennium, we can say that the older view of lactic acid and hypoxia is correct in the sense that anaerobic glycolysis, HLa production, and HLa accumulation are increased under conditions that engender O 2-limited oxidative phosphorylation in mitochondria. Section 12, Exercise: Regulation and Integration of Multiple Systems The mice were randomly divided into five groups (The incremental treadmill running protocol was based on a maximal running capacity test before the formal research. After the microplate was incubated in the dark for 10 min, the fluorescence of the samples was measured at 480 nm excitation and 520 nm emission using BioTek Synergy2 (USA) (Capture ELISA was used to measure the MPO-DNA complexes as described (IL-6 expression of mice serum was detected using the Quantikine ELISA kit (R&D, catalog number: M6000B) using instructions provided by the manufacturer. proportional to the extent to which the muscle had previously been In other words, it remains true that tissue hypoxia does indeed lead to increased HLa … OWestern and Northern blotting, immunofluorescence microscopy, and Haller et al. Lactic acidosis results from the accumulation of lactate and protons in the body fluids and is often associated with poor clinical outcomes. (Fifty male C57BL/6 mice (seven-week-old) were purchased from Shanghai JCJ laboratory animal center and were fed for 1-week acclimatization phase (environmental temperature 20–25°C with a 12 h light/dark cycle and free access to standard pellets and drinking water). The LA concentration stayed near 2 mmol/L during the first 75 min of running with the speed below 27 m/min. LA was detected through test strips using a Lactate Scout+ (EFK Diagnostics, Magdeberg, Germany). (First Published December 19, 2000; 10.1073/pnas.011387598). This study investigated NETs release, cell-free DNA (cf-DNA), and LA concentration in mice after 60 and 145 min of intensive, graded treadmill running. However, several studies during the past ≈30 years have presented accumulation, then what is? Front. Quantitative data were expressed as mean ± SEM (standard error of mean). 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