Assessment of the state of the athlete's molecular profile after short-term exercise
Description
Comprehensive analysis of indicators of the state of the body between training and recovery allows you to comprehensively evaluate various aspects of health, athletic performance and recovery. In this work, an assessment of the metabolomic profile of athletes was carried out, the immunological reaction of the athlete's body to food before exercise and 48 hours after exercise was studied. As a result, 15 amino acids and three hormones were identified, the plasma levels of which differed between training and recovery states. Additionally, an assessment of immunological reactions, or hyperreactivity, to food allergens was carried out by enzyme immunoassay. Probably, for athletes in the study sample, 48 hours is not enough for a complete recovery of the body.
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Supplementary 1 "MS analysis of the metabolic profile of athletes before exercise and after recovery": Data acquisition was performed on a high-resolution quadrupole time-of-flight (Q-TOF) Xevo G2-XS mass spectrometer (Waters, Inc., Ireland) equipped with a Z-spray ionization source and coupled with a UPLC Acquity H Class (Waters, Inc., Ireland) system. The instrument operated in a positive ionization source in ESI mode. Data were acquired in continual MSe over 50 – 700 m/z mass range within 1.875 s scan time using SONAR mode setting with a quadrupole peak width of 20 m/z. Precursor ions were decomposed with low collision energy level at 5 eV and high collision energy ramping between 12 – 40 eV using argon as a collision gas. Cone voltage was fixed at 16 V with source voltage offset of 21V and capillary voltage of 2.8 kV. Desolvation gas and cone gas (nitrogen) were adjusted to 700 L/h and 50 L/h, correspondingly, and source temperature was adjusted to 320oC. Lock-mass (warfarin, m/z = 309/1127) was acquired over the data collection for 70 ms interval of 10 s and averaged to three scans with mass window of 0.1 Da. Chromatographic separation was realized on an Acquity Premier HSS T3 column (2.1 x 150 mm, 8 µm particle size) heated at 50oC at 0.4 mL/min flow rate in a gradient of mobile phase A (water supplied with 0.1% formic acid and 0.005% heptafluorbutyric acid) and mobile phase B (acetonitrile with 0.1% formic acid) for 8.6 minutes starting from 2% of B for 0.4 minutes and increased to 18% of B at 0.65 minutes, then increased to 47% of B at 5.5 minutes and to 95% of B at 5.7 minutes and hold to 7.0 minutes. Then the gradient was returned to initial condition at 7.2 minutes and the column was equilibrated for the next 1.4 minutes. Supplementary 1 "Results of determination of hyperreactive reaction to food allergens mediated by immune processes": Immunological reactions were assessed by solid-phase non-competitive indirect enzyme immunoassay using a commercial Reagent Kit for semi-quantitative enzyme immunoassay for allergen-specific IgG antibodies (LLC NPO Immunoteks, Stavropol, Russia). This Kit uses monoclonal anti-IgG antibodies included in the peroxidase conjugate, capable of detecting antibodies of the immunoglobulin G class in human serum/plasma, which interact affinoly with allergens sorbed on the surface of a polystyrene tablet. The unbound components of the analyzed sample and the excess of the conjugate are washed off, and the activity of peroxidase in the immune kits is determined using a chromogenic substrate (tetramethylbenzidine). One Kit is designed to determine the content of specific IgG in eight test samples to 22 allergens. ELISA was carried out on a Multiskan FC microplate photometer (Thermo Scientific, USA) using the instructions for the Reagent Kit provided by the manufacturer.