There is limited information on skeletal muscle properties in women with unloading and countermeasure programs to protect the unloading-induced atrophy. The current investigation tested the hypothesis that a concurrent aerobic and resistance exercise training program would preserve size and contractile function of slow- and fast-twitch muscle fibers. A secondary objective was to test the hypothesis that a leucine-enriched high-protein diet would partially attenuate single fiber characteristics. Vastus lateralis muscle biopsies were obtained before and on day 59 of bed rest from a control (BR; n = 8), nutrition (BRN; n = 8), or exercise (BRE; n = 8) group. Single muscle fibers were studied for diameter, peak force (P(o)), contractile velocity, and power. Those in the BR group had a decrease (P < 0.05) in myosin heavy chain (MHC) I diameter (-14%), P(o) (-35%), and power (-42%) and MHC IIa diameter (-16%) and P(o) (-31%; P = 0.06) and an increase (P < 0.05) in MHC hybrid fibers. Changes in size and function of MHC I (-19 to -44%) and IIa (-21% to -30%) fibers and MHC distribution in BRN individuals were similar to results in the BR group. In BRE conditions, MHC I and IIa size and contractile function were preserved during bed rest. These data show that the concurrent exercise program preserved the myocellular profile of the vastus lateralis muscle during 60-day bed rest. To combat muscle atrophy and function with long-term unloading, the exercise prescription program used in this study should be considered as a viable training program for the upper leg muscles, whereas the nutritional intervention used cannot be recommended as a countermeasure for skeletal muscle.