Change n weight from baseline to 6 months
Change from baseline weight (kg) at 6 months. For weight assessment, a Tanita 780PMA model precision scale will be used. It will always be carried out with the patient in underwear, standing on the scale in a standard anatomical position without using any extra point of support that could modify the measurement.
Patients will be measured with the direct reading Harpenden Stadiometer with a wall-mounted indicator counter, measuring 600-2100 mm, approved by the University of London Institute of Child Health; standing on the height rod, without wearing shoes.
Change on body mass index (BMI) from Baseline to 6 months
To calculate changes in BMI is necessary to determine weight (kilograms) and height (meters).
The body mass index (BMI) or Quetelet index (Weight/Height²) will be stratified according to the International reference values of Cole and Bellizi to classify obesity.
Change of body composition from Baseline to 6 months
Bioimpedance test will be performed to assess changes on body composition, with emphasis on fat mass, using a Tanita 780PMA model precision scale.
Dietary intake and eating habits
Assessment of changes on dietary intake will be evaluated at the beginning, 3 and 6 months after the intervention with three 24-hour reminders separated by at least one week in accordance with the EFSA recommendations for this type of study, all of them previously validated. At the beginning of the study, a questionnaire on the frequency of food consumption will be carried out.
Physical activity
Assessment of changes on physical activity will be assessed at the beginning, 3 and 6 months after the intervention through the PAQ-C Physical Activity questionnaire
Sedentary activities
Assessment of the level of sedentary activities will be evaluated at the beginning, 3 and 6 months after the intervention through an ad hoc questionnaire on time spent watching television and use of computers, video games and consoles, validated for young children and school children, as well as other socioeconomic determinants
Hemogram
Changes on blood count will be analyzed on an automated analyzer
Blood glucose
Changes on blood glucose will be analyzed by the glucose oxidase method on an automated analyzer (Roche-Hitachi Modular P and D Autoanalyser; Roche Laboratory Systems, Mannheim, Germany)
Blood insulin
Changes on plasma insulin will be analyzed by radioimmunoassay (RIA) using an automated microparticle analyzer. (AxSYM; Abbott Laboratories, Abbott Park, IL, USA).
Insulin resistance
Changes on insulin resistance (IR) will be calculated by homeostatic model assessment of IR (HOMA-IR).
Serum triacylglycerides
Changes on serum, triglycerides will be measured using an automatic analyzer ( Roche-Hitachi Modular P and D Autoanalyser; Roche Laboratory Systems, Mannheim, Germany).
Serum cholesterol
Changes on serum total cholesterol, high-density lipoprotein cholesterol (HDLc) and low-density lipoprotein cholesterol (LDLc), as well as Apoprotein A and B, and Lipoprotein A1 will be measured using an automatic analyzer ( Roche-Hitachi Modular P and D Autoanalyser; Roche Laboratory Systems, Mannheim, Germany).
Endothelial function biomarkers in blood
Simultaneous detection of endothelial adhesion molecules VCAM-1, ICAM, E-selectin and myeloperoxidase will be performed in children's plasma, using MULTIPLEX assay kits and Luminex xMAP detection technology, which allows us to quantitatively determine several biomarkers using a very small amount of sample (25-50 µl of plasma or serum). The HCVD2MAG-67K kit will be used to determine the adhesion molecules.
Antioxidant glutathione reductase activity in red blood cells
Assessing changes of glutathione reductase activity will be carried out using standardized spectrophotometric methods.
Antioxidant glutathione peroxidase activity in red blood cells
Assessing changes of glutathione peroxidase activity will be carried out using standardized spectrophotometric methods.
Antioxidant superoxide dismutase activity in red blood cells
Assessing changes of superoxide dismutase activity will be carried out using standardized spectrophotometric methods.
Antioxidant catalase activity in red blood cells
Assessing changes of catalase activity will be carried out using standardized spectrophotometric methods.
Antioxidant alpha-tocopherol, beta-carotene, retinol and Coenzyme Q in plasma
Assessing changes of plasma antioxidants: alpha-tocopherol, beta-carotene, retinol, and coenzyme Q will be analysed by HPLC-MS in one single inyection
Urine endothelin-1
Assessing changes of endothelin-1 will be analyzed in plasma using an ELISA kit.
Analysis of oxLDL in plasma
Assessing changes of oxidized LDL from the baseline to the end of each treatment, by using an ELISA kit
F2-Isoprostanes in urine
Assessing changes of F2-Isoprostanes will be carried out using an ELISA kit.
8-hydroxy-deoxy-Guanosine in urine
Assessing changes of 8-hydroxy-deoxy-Guanosine will be carried out using an ELISA kit.
Metagenomics
Assessing changes of fecal microbiome profile from the baseline to the end of each treatment. Stool DNA will be isolated with the QIAamp DNA stool mimi kit. Amplification of variable region V3-V1 of 16S gen will be sequenced using the Illunina Next Generation Sequencing MiSeg.
Plasma metabolomic analysis
A liquid chromatography platform coupled to a spectrophotometer will be used mass (LC/MS) to determine metabolic profiles by targeted analyses
Urine metabolomic analysis
A liquid chromatography platform coupled to a spectrophotometer will be used mass (LC/MS) to determine metabolic profiles by targeted analyses
Genotyping
Genotyping of the main candidate genes related to the development of arterial hypertension described in GWAS studies (ACE, ADRB1, AGT, PHACTR1, EDN1, ATP13A3) will be carried out in the Department of Biochemistry and Molecular Biology II of the University of Granada.