We conducted a cross-sectional analysis of baseline data (collected on February-March and October-November 2022) from the e-MOVI study aiming to study physical activity and diet behaviors, lifestyle and cardiovascular risk that occur during childhood. The study included 904 schoolchildren aged 9 to 12 years old from 8 public primary schools in the province of Cuenca, Spain.
The study protocol was approved by the Clinical Research Ethics Committee of the Hospital Virgen de la Luz in Cuenca (REG: 2019/PI1519). After the Board of Governors of each school approved the study, a letter was sent to the parents of all 4th, 5th and 6th graders inviting them to a meeting. At this meeting, we explained the objectives of the study and asked for written approval for their children's participation. All procedures performed in this study were in accordance with the Declaration of Helsinki and its later amendments or comparable ethical standards for experiments involving humans.
Height and weight were measured twice and averaged for analysis. Height was measured to the nearest millimetre using a wall-mounted stadiometer (SECA 222, Vogel and Halke, Hamburg, Germany), with the children standing straight against the wall without shoes, to align the spine with the stadiometer. The head was positioned so that the chin was parallel to the floor. Weight was measured to the nearest 100 g using a calibrated digital scale (SECA 861; Vogel & Halke, Hamburg, Germany) with the children lightly dressed and without shoes. Body mass index was calculated as weight in kilograms divided by the square of height in meters (kg/m2) using the means of the weight and height measurements.
Steps per day were measured by a Xiaomi Mi Band 3 Smart wristband, which presents a good validity for step count [15]. The mean of the steps per day in the two weeks following the baseline measurements was used. Only data from participants whose wristbands had records of at least 4 days per week, with at least 1 weekend day, were considered. The steps per day were recorded by the children in a daily steps log that was collected weekly at the school by a member of the research team.
The 20-m shuttle run test was used to assess the CRF. Participants ran between two lines separated by 20 m while keeping pace with audio signals emitted by a pre-recorded compact disc. The initial speed was 8.5 km/h and increased by 0.5 km/h every minute. Children were encouraged to keep running as long as possible during the test. When children failed to follow the pace in two consecutive intervals, they were asked to stop, and the stage completed (full laps) was recorded. Leger’s formula was used to estimate maximal oxygen consumption (VO2max) [31.025 + (3.238 × velocity) − (3.248 × age) + (0.1536 × age × velocity)] as an indicator of CRF.
Health-related quality of life was evaluated using the validated Spanish version of the KIDSCREEN-27 questionnaire [16], which consists of 27 items, referring to the previous week, assessing 5 domains of HRQoL: physical well-being, psychological well-being, parental relationships and autonomy, social support and peers, and school environment. T scores of each dimension were calculated according to the manual, using a mean of 50 and a standard deviation (SD) of 10, to establish normality for children [17] and in this way to obtain a single total score or global index of HRQoL from the means of the five dimensions, where a higher score represents better HRQoL.
Mother’s education was measured by using a validated questionnaire about the maximum level of education achieved by parents [18]. Responses were categorised into four levels: “no studies or primary studies”, “secondary studies”, “high school”, and “university studies”.
Before proceeding with the analysis, the normality of the distribution of the continuous variables was checked using statistical methods (Kolmogorov‒Smirnov) and graphs (normal probability plots). Differences in basic characteristics between girls and boys were tested using Student´s t tests for continuous variables and chi-square tests for categorical variables. Homogeneity of variances was assessed with the Levene test.
Partial correlation coefficients adjusted by sex, age, and mother’s education level (considered the familial socioeconomic indicator more closely related to HRQoL and lifestyle behavior [19, 20]) were calculated to examine the relationship between steps per day, CRF, and HRQoL.
A multivariable linear regression model was used to estimate the linear association between 1000 steps/day increment and CRF, adjusting for sex, age, and mother’s education level. Furthermore, to explore the relation between steps per day and CRF, a locally weighted scatterplot smoothing (LOESS) regression was used. According to the shape of the LOESS regression, steps per day were categorised into 3 categories: <9,000 steps/day, 9,000–12,000 steps/day and > 12,000 steps/day. Analysis of covariance (ANCOVA) models were used to test the mean differences in CRF and HRQoL global score by steps per day categories (model 0), controlling for sex, age, and mother’s education level (model 1) and CRF (only when HRQoL global score was the dependent variable, model 2). Differences in the mean scores of each HRQoL by steps per day categories were examined using a multivariate analysis of covariance (MANCOVA), controlling for the above-described potential confounders. Second- and third-order interaction terms were evaluated in ANCOVA and MANCOVA. Pairwise post hoc hypotheses were tested using the Bonferroni correction for multiple comparisons.
Mediation analyses adjusted by sex, age, and mother’s education level were conducted to examine whether CRF mediates the association between steps per day and HRQoL (domains and global score) using the PROCESS SPSS Macro, version 3.5, selecting Model 4 and 5000 bias-corrected bootstrap samples [21]. In these analyses, the first equation (path a) corresponded to the regression coefficients of the mediating variable (CRF) on the independent variable (steps per day), and the second equation (path c, total effect) was the regression coefficient of the outcome (HRQoL) on the independent variable. Path b represents the regression coefficient of the outcome controlling for the independent variable. The relationship of the independent variable with the outcome and the mediator simultaneously was estimated in the third equation (path c’, direct effect). To test the statistical significance of the mediation effect in the parametric approach, we used the indirect effect (path a * path b), which indicates the change in HRQoL per unit change in steps per day that is mediated by CRF. Following the Hayes recommendation [22], the complete and partial mediation concepts were not used in this study.
All statistical analyses were performed using IBM SPSS Statistics software (Version 28.0; IBM Corp., Armonk, NY, USA), and p < 0.05 was considered to indicate significance.