The Push-Pull Effect (PPE) is a physiological phenomenon defined as a reduction of +Gz tolerance induced by a previous exposure to a -Gz acceleration, that may lead to loss of consciousness. Aim of this study was to evaluate, for the first time, the cardiac rhythm changes associated with PPE during real flights. Data were collected in 3 pilots during flights on the Aermacchi MB- 339-CD aircraft. In each flight, lasting about 60 minutes, ECG, respiration and 3D accelerations were recorded by a new smart garment (the MagIC System). The flight protocol included a maneuver eliciting a reference +5Gz acceleration for 15 seconds (Ref+5G), followed, after a while, by a push-pull maneuver with a profile characterized by a 5-s acceleration at -1Gz (PP-1G) and, with a 1G/s onset, by 15 seconds at +5Gz (PP+5G), so to induce the push-pull gravitational stress. From each ECG recording, a beat-by-beat RR Interval (RRI) series was derived. RRI mean, standard deviation (SDNN) and the RRI Root Mean Square of Successive Difference (RMSSD) were estimated in each pilot during the Ref+5G and the PP+5G maneuvers. As compared with Ref+5G, all pilots displayed significant reductions in RRI mean, SDNN and RMSSD during PP+5G. These findings are compatible with a PP-induced enhancement in the sympathetic drive to the heart -as shown by the reduction in RRI mean and SDNN- and a concomitant deactivation of the parasympathetic control as shown by the reduction in RMSSD.