Abstract: The autonomic nervous system (ANS) plays a crucial role in physiological regulation by balancing sympathetic and parasympathetic activity. Heart rate variability (HRV) is a well- established method for assessing autonomic function. This pilot study aimed to evaluate the effects of NanoVi® therapy on autonomic regulation using HRV analysis. Objective: This study explored whether a 15-minute session of NanoVi® could enhance HRV parameters, particularly those related to parasympathetic tone and stress response. Methods: Thirty-nine (n=39) participants underwent baseline HRV measurement (Group 1) followed by a 15-minute NanoVi® therapy session, after which HRV was reassessed (Group 2). HRV parameters were measured using the ANS Analysis device (Creating Health, Germany). Key autonomic markers analyzed included: RMSSD (Root Mean Square of Successive Differences) – a measure of parasympathetic activity; Stress Index (SI, Baevsky method) – an indicator of sympathetic activity; Alpha-1 value (DFA 1) – an index of overall autonomic regulation and coordination. The Software GraphPad Prism, v10.0 (La Jolla, CA, USA) was used for the analysis. Normality assessed with the Shapiro-Wilk test. Wilcoxon matched-pairs signed-rank test used for non-parametric comparisons. Results : Parasympathetic Activity (RMSSD) increased by 19.4% (14.19 to 16.95, p = 0.0001) (Figure 1). Sympathetic Activity (Stress Index) decreased by 45.7% (403.74 to 201.22, p < 0.0001) (Figure 2). Autonomic Regulation (Alpha-1 value) increased by 4.5% (1.323 to 1.384, p = 0.0495) (Figure 3). Conclusion: This pilot study suggests that a single 15-minute session of NanoVi® therapy significantly enhances parasympathetic tone, reduces sympathetic overactivity, and improves autonomic coordination. These findings indicate that NanoVi® may support autonomic balance and stress regulation, potentially Page 2 benefiting individuals seeking enhanced recovery, wellness, or performance optimization. Future research with larger sample sizes, control groups, and longterm intervention studies is warranted to confirm and expand upon these results.