Heart rate variability (HRV) is a non-invasive indicator of autonomic nervous system function. HRV recordings show artefacts due to technical and/or biological issues. The Kubios software is one of the most used software to process HRV recordings, offering different levels of threshold-based artefact correction (i.e., Kubios filters). The aim of the study was to analyze the impact of different Kubios filters on the quantification of HRV derived parameters from short-term recordings in three independent human cohorts. A total of 312 participants were included: 107 children with overweight/obesity (10.0 ± 1.1 years, 58% men), 132 young adults (22.2 ± 2.2 years, 33% men) and 73 middle-aged adults (53.6 ± 5.2 years, 48% men). HRV was assessed using a heart rate monitor during 10–15 min, and the Kubios software was used for HRV data processing using all the Kubios filters available (i.e., 6). Repeated-measures analysis of variance indicated significant differences in HRV derived parameters in the time-domain (all p < 0.001) across the Kubios filters in all cohorts, moreover similar results were observed in the frequency-domain. When comparing two extreme Kubios filters, these statistical differences could be clinically relevant, e.g. more than 10 ms in the standard deviation of all normal R-R intervals (SDNN). In conclusion, the results of the present study suggest that the application of different Kubios filters had a significant impact on HRV derived parameters obtained from short-term recordings in both time and frequency-domains.

Accelerometers’ accuracy for sedentary time (ST) and moderate-to-vigorous physical activity (MVPA) classification depends on accelerometer placement, data processing, activities, and sample characteristics. As intensities differ by age, this study sought to determine intensity cut-points at various wear locations people more than 70 years old. Data from 59 older adults were used for calibration and from 21 independent participants for cross-validation purposes. Participants wore accelerometers on their hip and wrists while performing activities and having their energy expenditure measured with portable calorimetry. ST and MVPA were defined as ≤1.5 metabolic equivalents (METs) and ≥3 METs (1 MET = 2.8 mL/kg/min), respectively. Receiver operator characteristic (ROC) analyses showed fair-to-good accuracy (area under the curve [AUC] = 0.62–0.89). ST cut-points were 7 mg (cross-validation: sensitivity = 0.88, specificity = 0.80) and 1 count/5 s (cross-validation: sensitivity = 0.91, specificity = 0.96) for the hip; 18 mg (cross-validation: sensitivity = 0.86, specificity = 0.86) and 102 counts/5 s (cross-validation: sensitivity = 0.91, specificity = 0.92) for the non-dominant wrist; and 22 mg and 175 counts/5 s (not cross-validated) for the dominant wrist. MVPA cut-points were 14 mg (cross-validation: sensitivity = 0.70, specificity = 0.99) and 54 count/5 s (cross-validation: sensitivity = 1.00, specificity = 0.96) for the hip; 60 mg (cross-validation: sensitivity = 0.83, specificity = 0.99) and 182 counts/5 s (cross-validation: sensitivity = 1.00, specificity = 0.89) for the non-dominant wrist; and 64 mg and 268 counts/5 s (not cross-validated) for the dominant wrist. These cut-points can classify ST and MVPA in older adults from hip- and wrist-worn accelerometers.

A high inter-day reliability is a key factor to analyze the magnitude of change in resting metabolic rate (RMR) after an intervention, and the impact of using different methods for data analysis is not known. The aims of this study were: i) to analyze the impact of methods for data analysis on RMR and respiratory exchange ratio (RER) estimation; ii) to analyze the impact of methods for data analysis on inter-day RMR and RER reliability; iii) to compare inter-day RMR and RER reliability across methods for data analysis in participants who achieved steady state (SS) vs. participants who did not achieve SS. Seventeen young healthy adults completed two 30-min indirect calorimetry (IC) measures on two consecutive mornings, using two metabolic carts each day. Two methods for data analysis were used: i) Selection of a predefined time interval (TI) every 5 min (1-5 min, 6-10 min, 11-15 min, 16-20 min, 21-25 min, 26-30 min); and TI representing the whole measurement period (0-30 min, 5-30 min, 5-25 min); and ii) Methods based on the selection of the most stable period (SSt methods) (3 min SSt, 4 min SSt, 5 min SSt, 10 min SSt). Additionally, participants were classified as those achieving SS (CV < 10% for VO2, VCO2 and VE, and CV < 5% for RER) and those who did not. RMR and RER measurements were lower when following SSt methods than when following TI methods (all P < 0.01). Although no significant differences were found between different lengths of SSt, 5 min SSt presented the lowest RMR. There were no differences on the inter-day reliability across methods for data analysis (TI and SSt) (all P > 0.2), and there was no systematic bias when comparing RMR and RER day 1 and day 2 measurements (all P > 0.1). Inter-day reliability was similar in individuals who achieved the SS and individuals who did not achieve it. The results were consistent independently of the metabolic cart used. The 5 min SSt approach should be the method of choice for analyzing IC measures with metabolic carts. However, achieving SS should not be an inclusion criterion in an IC study with young healthy adults.

Brown adipose tissue (BAT) activity is induced when humans are exposed to cold. Therefore, cold exposure prior to the 18F-FDG-PET/CT scan is used as a tool to quantify BAT. Several cooling protocols, including fixed and personalised ones are currently in use. The aim of the present study was to determine the effect of a new personalised cooling protocol where the shivering threshold was measured on a separate day, on BAT volume and activity in young adults. A total of 47 adults (n=28 women) aged 22±2 years participated in the study. We determined participants’ shivering threshold (visually and self-reported) using a water perfused cooling vest in an air-conditioned cold room. 48-72 hours later, participants wore the cooling vest set at ~4ºC above the shivering threshold for 60 minutes prior to injection of 18F-FDG and ~5ºC above the shivering threshold for ~60 minutes after injection, until PET/CT scan. We quantified BAT following BARCIST 1.0 recommendations. We identified 40 participants (85%, n=25 women) as PET+ and 7 (n=3 women) as PET-. The PET+ group presented significantly higher BAT volume and activity than PET- group (all P<0.05). PET+ women had higher BAT mean activity than PET+ men (SUVmean: 5.0±1.6 vs. 3.6±0.9 g/ml respectively, P=0.003), and there were no significant sex differences in BAT volume (P=0.161). A total of 9 out of 47 participants did not shiver during the shivering threshold test. Our findings are similar to previous cold-stimulated human BAT studies; therefore, we conclude that our personalised cooling protocol is able to activate BAT in young adults.

The validity of most commercially available metabolic cart is mostly unknown. Thus, we aimed to determine the accuracy, precision, within-subject reproducibility, and concordance of RMR and RER measured by four commercially available metabolic carts [Cosmed Q-NRG, Vyaire Vyntus CPX, Maastricht Instruments Omnical, and Medgraphics Ultima CardiO2]. Further, we studied whether a previously proposed simulation-based post-calorimetric calibration of cart readouts [individual calibration control evaluation (ICcE)] modify the RMR and RER reproducibility and concordance. Three experiments simulating different RMR and RER by controlled pure gas (N₂ and CO₂) infusions were conducted on 5 non-consecutive days. Moreover, 30-min methanol burns were per- formed on 3 non-consecutive days. Lastly, the RMR and RER of 29 young non-ventilated adults (11 women; 25 ± 4 years-old; BMI: 24.1 ± 3.2 kg/m²) were assessed twice using each instrument, 24 hours apart, under standardized conditions. The Omnical presented the lowest measurement error for RER (Omnical = 1.7 ± 0.9%; Vyntus = 4.5 ± 2.0%; Q-NRG = 6.6 ± 1.9%; Ultima = 6.8 ± 6.5%) and EE (Omnical = 1.5 ± 0.5%; Q- NRG = 2.5 ± 1.3%; Ultima = 10.7 ± 11.0%; Vyntus = 13.8 ± 5.0%) in all in vitro experiments (controlled pure gas infusions and methanol burns). In humans, the 4 metabolic carts provided discordant RMR and RER estimations (all P < 0.001). No differences were detected in RMR within-subject reproducibility (P = 0.058; Q-NRG inter-day coefficient of variance = 3.6 ± 2.5%; Omnical = 4.8 ± 3.5%; Vyntus = 5.0 ± 5.6%; Ultima = 5.7 ± 4.6%), although the Ultima CardiO2 provided larger RER inter-day differences (4.6 ± 3.5%) than the other carts (P = 0.001; Omnical = 1.9 ± 1.7%; Vyntus = 2.1 ± 1.3%; Q-NRG = 2.4 ± 2.1%). The ICcE procedure did not modify the RMR or RER concordance and did not reduce the inter-day differences in any of the carts. The 4 metabolic carts provided discordant measurements of RMR and RER. Overall, the Omnical provides more accurate and precise estimations of RMR and RER than the Q-NRG, Vyntus and Ultima CardiO2, and might be considered the best for assessing RMR and RER in non-ventilated humans. Finally, our results do not support the use of an ICcE procedure.