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INTRODUCTION Current exercise countermeasures on the International Space Station (ISS) include treadmill running, cycle ergometry, and resistive exercise, which are used to protect crewmember health and performance during long-duration spaceflight. However, exploration vehicles for Artemis missions to the Moon and beyond will have volume and power restrictions, requiring exercise hardware to have a smaller footprint and use fewer resources. Thus, recent efforts have focused on developing exercise devices that provide both aerobic and resistive capabilities on one platform without including a treadmill. The European Enhanced Exploration Exercise Device [E4D] is one such apparatus. It is critical to validate the efficacy of using exploration-focused exercise modalities to preserve muscle strength, aerobic fitness, bone density, and sensorimotor performance. Thus, the aim of this study is to determine the effectiveness of using nominal ISS exercise devices for an entire mission compared to exploration-forward exercise modalities to determine if a treadmill is required to maintain current levels of protection during long-duration missions. METHODS Crewmembers are assigned to one of three groups: 1) Controls (n ≥ 40), who partake in nominal exercise on the ISS, including running on the Treadmill with Vibration Isolation and Stabilization 2 (T2), cycling on the Cycle Ergometer with Vibration Isolation and Stabilization (CEVIS), and strength training on the Advanced Resistive Exercise Device (ARED); 2) Active Group 1, who exercise on the CEVIS and ARED only (n = 8); and 3) Active Group 2, who perform aerobic and resistive exercise on the E4D only (n = 8). For Active Group 1, nominal exercise on T2 will be replaced with corresponding exercise on CEVIS. For Active Group 2, a dedicated exercise prescription will be designed to maximize the capabilities of the E4D to include resistive exercise, cycle ergometry, rowing, and rope pulling. Crewmembers in both Active groups are not permitted to perform treadmill exercise for the entirety of their flight. Health and performance markers, including bone mineral density (dual-energy x-ray absorptiometry [DXA]), body composition (DXA), cardiovascular fitness (cycle VO2peak), muscle strength and endurance (isometric/isokinetic testing, power endurance testing), sensorimotor performance (sit-to-stand, obstacle course), postural control (computerized dynamic posturography), and blood and urine biochemical markers of bone metabolism will be assessed before, during, and after spaceflight. RESULTS Fifteen subjects (4 Active [CEVIS + ARED], 11 Control) have been recruited. Data collection is ongoing. CONCLUSIONS This study will assess the efficacy of using exploration exercise modalities, including removing the treadmill exercise capability or exclusively using the E4D, compared to the nominal ISS exercise regimen across an entire mission on bone, muscle, aerobic, and sensorimotor health and performance. Findings from this study will help provide a recommendation on whether these exploration exercise modalities can sufficiently protect against physiological deconditioning during spaceflight or whether a treadmill may be required to maintain current levels of protection during future exploration class spaceflight missions. Supported by the NASA Human Research Program and NASA Exploration Capabilities