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Objective: Ambulatory venous pressure (AVP) is the drop of pressure observed in the superficial veins of the lower leg during movement. This phenomenon has been linked to the function of the calf muscle pump (CMP) and the competence of venous valves. Nevertheless, the concept of the CMP function remains controversial. This study aimed to elucidate the association between lower leg muscles activity, changes in pressure in distinct venous segments, and lower extremity arterial blood supply in healthy subjects during various types and intensities of exercise., Methods: Twelve legs of nine healthy volunteers were enrolled in the study. Continuous pressure (intramuscular vein [IV] and three great saphenous vein [GSV] points) and surface electromyography data (gastrocnemius and anterior tibial [ATM] muscles) were recorded during treadmill walking, running, and plantar flexion exercises. The pressure gradient (ΔP, mmHg) between adjacent points of measurement was calculated. Minute unit power of muscle pump ejection and suction (N E , and N S , MPa/min) were calculated and compared with the arterial blood supply of the lower extremity (LBF, L/min)., Results: ΔP demonstrated a consistent pattern of changes during walking and running. In GSV, the ΔP was observed to be directed from the thigh to the mid-calf (retrogradely) and from the ankle to the mid-calf (anterogradely) throughout the entire stride cycle. However, its value decreased with increasing stride cycle frequency. The dynamics of ΔP between the IV and GSV were as follows: It was directed from the IV to GSV during gastrocnemius contraction and was reversed during anterior tibial muscle contraction and gastrocnemius relaxation (swing phase). LBF, N E , and N S demonstrated similar exponential growth with increasing stride frequency during walking and running., Conclusions: During natural locomotion, the muscle pump acts as a flow diverter pump, redirecting the flow of blood from the superficial veins to the intramuscular veins via the perforating veins. During ambulation, the pressure in the superficial venous network depends upon the capacity of the muscle pump to provide output that matches the changes in arterial blood flow., Competing Interests: Disclosures None., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)