The COVID-19 pandemic has marked a turning point in the working habits of many office employees. The evaluation of the impact of working at home rather than in office has been mostly limited to the administration of questionnaires. Recently, the assessment of cardiac autonomic control profile (CACP) has been suggested as an early, non-specific, screening tool in occupational medicine. The aim of this study is to determine whether different working conditions may modify CACP. Seventeen healthy office active employees (age: 38±7 yrs, 9 males, 8 females) were studied by means of 24-hour Holter electrocardiogram while alternating working at home (SMART) and in the office (OFFICE). The perceived level of stress was assessed via a visual analogue scale (VAS). Heart period was derived as the time distance between two consecutive R-wave peaks (RR). Parametric power spectral analysis was carried out over sequences of 250 RRs and iterated, with 80% overlap, over segments of 5000 RRs during daytime (DAY) and nighttime (NIGHT). Median values of variance (σ2RR) and the absolute power of RR series in high frequency (HF, 0.15-0.4 Hz) band (HFaRR) were computed. SMART and OFFICE conditions led to similar habits as measured via slept hours per night and working hours per day. The VAS score was higher in OFFICE than SMART (6.8±1.1 vs 4.8±1.5). During NIGHT, σ2RR was higher in SMART compared to OFFICE (4298±4112 vs 2877±2360 ms2). A similar tendency was observed in the HFaRR power (SMART: 1406±2826 ms2; OFFICE: 871±1280 ms2). We conclude that during NIGHT the vagal modulation is higher in SMART compared to OFFICE together with a lower VAS, that in turn may reflect beneficial effects for the cardiovascular system and the overall state of well-being.