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Prolonged sedentary behaviour (SB) i.e. longer bouts, is suggested to have a range of negative health effects, independent of habitual light and medium-to-vigorous physical activity (LIPA or MVPA). Any effect on musculoskeletal size, architecture or morphology has seldom been reported in older adults. Moreover, no study has yet determined if any association would persist following adjustment for covariates. Therefore, the aim of the present study was to investigate the associations between SB, and properties of the Gastrocnemius Medialis (GM) muscle, in a cross-sectional sample of older adults using compositional data analysis. 105 healthy older adults (73±6y) wore a thigh mounted tri-axial accelerometer for seven consecutive days, and underwent ultrasound [e.g. muscle length (Lm), anatomical cross-sectional area (ACSA), muscle volume (VM), fascicle length (LF), & physiological cross-sectional area (PCSA)], body composition (e.g. DEXA) and health (e.g. medical history) assessments. In-unadjusted models, SB time was negatively associated with ACSA at 75% of Lm (R2adj = 0.085), VM (R2adj = 0.020), and PCSA (R2adj = 0.039). Standing was positively associated with pennation angle (R2adj = 0.110), which persisted following co-variate adjustment (R2adj = 0.296). In fully adjusted models, both SB & LIPA time were associated with ACSA at 75% of Lm (Both R2adj = 0.393). Standing and light activity time were also associated with LF, VM, & PCSA (R2adj 0.116-0.573). In fully adjusted models, SB pattern parameters (i.e. the manner in which sedentary behaviour is accumulated daily throughout waking hours such as the timing, duration and frequency of sedentary bouts), were associated with GM muscle properties (R2adj 0.156-0.564) including LM, LF, and VM. The pattern, rather than accumulated daily SB time, was associated with the size and architecture of the GM. Our results suggest that regardless of co-existing habitual physical activities, SB bouts should be kept short and freque

Sedentary behaviour (SB) and physical activity (PA) have been shown to be independent modulators of healthy ageing. We thus investigated the impact of activity monitor placement on the accuracy of detecting SB and PA in older adults, as well as a novel random forest algorithm trained on data from older persons. Four monitor types (ActiGraph wGT3X-BT, ActivPAL3c VT, GENEActiv Original, and DynaPort MM+) were simultaneously worn on five anatomical sites during ten different activities by a sample of twenty older adults (70.0 (12.0) years; 10 women). The results indicated that collecting metabolic equivalent (MET) data for 60 s provided the most representative results, minimising variability. In addition, thigh-worn monitors, including ActivPAL, Random Forest, and Sedentary Sphere—Thigh, exhibited superior performance in classifying SB, with balanced accuracies ≥ 94.2%. Other monitors, such as ActiGraph, DynaPort MM+, and GENEActiv Sedentary Sphere—Wrist, demonstrated lower performance. ActivPAL and GENEActiv Random Forest outperformed other monitors in participant-specific balanced accuracies for SB classification. Only thigh-worn monitors achieved acceptable overall balanced accuracies (≥80.0%) for SB, standing, and medium-to-vigorous PA classifications. In conclusion, it is advisable to position accelerometers on the thigh, collect MET data for ≥60 s, and ideally utilise population-specific trained algorithms.