Antihypertensive and Cardioprotective Evaluation of a Functional Olive Oil

Study Overview
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Brief Summary
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The general objective of this study is to evaluate the effect of a nutritional intervention with an extra virgin olive oil (EVOO) rich in bioactive compounds (polyphenols) and a functional oil of adequate organoleptic quality, prepared with the same EVOO rich in polyphenols and also enriched in triterpenic acid compounds from the olive itself, and demonstrate its antihypertensive and cardioprotective effect in children and adolescents (between 6-18 years) at risk of untreated hypertension.

Detailed Description
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A clinical study of nutritional intervention, randomized, adjusted by randomization blocks that consider age and sex, and parallel duration of 6 months, in 75 subjects (25 per randomization group) is proposed. The study will be carried out at the UGC of Pediatrics of the Reina Sofía University Hospital in Córdoba, as well as at the Institute of Nutrition and Food Technology of the University of Granada. The effects of an optimized oil rich in polyphenols and obtained by mixing varieties rich in said bioactive compounds (approx. 490 ppm), and a functional oil prepared with the same optimized oil and enriched with triterpenic acids obtained from the olive itself (approx. 490 ppm of polyphenols and 380 ppm of triterpenic acids). The study will be carried out in pediatric patients with high levels of untreated blood pressure, in which the effect of oils on blood pressure and their action on biomarkers of cardiovascular risk and on the intestinal microbiota will be evaluated, depending on the genotype specific to each subject. In addition, the presence of minor olive oil compounds present in the plasma will be analyzed (metabolomic analysis). In this way, at the end of the project, verified scientific information will be available on the enriched product and its effects on cardiovascular protection, and on possible mechanisms of action, as well as on its palatability and acceptance. The general objective of this study is to evaluate the effect of a nutritional intervention with an EVOO rich in bioactive compounds (polyphenols) and a functional oil of adequate organoleptic quality, prepared with the same EVOO rich in polyphenols and also enriched in triterpenic acid compounds from the olive itself, and demonstrate its antihypertensive and cardioprotective effect in children and adolescents (between 6-18 years) at risk of untreated hypertension. As specific objectives will be determined: To evaluate the effect of olive oils on blood pressure levels To evaluate the effect of olive oils on the levels of other components of the metabolic syndrome. To evaluate the effect of olive oils on the antioxidant defense system To evaluate the effect of olive oils on biomarkers of cardiovascular risk, including markers of inflammation and endothelial damage. To evaluate the effect of olive oils on the intestinal microbiota. To evaluate the effect of olive oils on metabolomic changes in plasma. To analyze the specific genotype of each volunteer and its relationship with the clinical effects found.

Official Title
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Antihypertensive and Cardioprotective Clinical Evaluation of a Functional Olive Oil Enriched in Bioactive Compounds From the Olive Tree

Conditions
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Hypertension

Intervention / Treatment
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* Dietary Supplement: Optimized oil group
* Dietary Supplement: Functional oil group
* Dietary Supplement: Control group


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Age between 6 and 18 years Percentile levels compatible with arterial hypertension or very high risk (SBP and/or DBP over P90) Without previous medication for hypertension Acceptance to participate by signing the informed consent Exclusion Criteria: Children under 6 years of age Absence of hypertension Children who require or receive some type of medication to control blood pressure, glucose levels or dyslipidemia or any probiotic Not to sign the informed consent.

Ages Eligible for Study
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Minimum Age: 6 Years
Maximum Age: 18 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: Optimized oil group<br>Each participant will consume 25 mL/day during 6 months of an Optimized extra virgin olive oil obtained by mixing different varieties of oils rich in bioactive compounds (it contains approx. 490 ppm of bioactive compounds). | Dietary Supplement: Optimized oil group<br>* This is a controlled parallel study. The patient will receive 25 mL/day of three different types of olive oils with different amouns of bioactive compounds from the olive tree, that must be consumed throughout the day at breakfast or snack (depending on the habits of each child). The doses that they do not take must be returned to the hospital staff to assess adherence to the intervention. The families will receive the same type of oil so that they can cook with it during the study period. The control oil will be the same extra virgin olive oil washed to decrease bioactive components (approx. 125 ppm). The intervention will take place for 6 months.<br>|
| Experimental: Functional oil group<br>Each participant will consume 25 mL/day during 6 months of a Functional olive oil prepared with the same Optimized oil and enriched with triterpenic acids obtained from the olive tree (approx. 490 ppm of polyphenols and 380 ppm of triterpenic acids). | Dietary Supplement: Functional oil group<br>* This is a controlled parallel study. The patient will receive 25 mL/day of three different types of olive oils with different amouns of bioactive compounds from the olive tree, that must be consumed throughout the day at breakfast or snack (depending on the habits of each child). The doses that they do not take must be returned to the hospital staff to assess adherence to the intervention. The families will receive the same type of oil so that they can cook with it during the study period. The control oil will be the same extra virgin olive oil washed to decrease bioactive components (approx. 125 ppm). The intervention will take place for 6 months.<br>|
| Placebo Comparator: Control Group<br>Each participant will consume 25 mL/day during 6 months of a Control extra virgin olive oil prepared from the same Optimized oil washed to eliminate bioactive compounds (approx. 125 ppm of polyphenols). | Dietary Supplement: Control group<br>* This is a controlled parallel study. The patient will receive 25 mL/day of three different types of olive oils with different amouns of bioactive compounds from the olive tree, that must be consumed throughout the day at breakfast or snack (depending on the habits of each child). The doses that they do not take must be returned to the hospital staff to assess adherence to the intervention. The families will receive the same type of oil so that they can cook with it during the study period. The control oil will be the same extra virgin olive oil washed to decrease bioactive components (approx. 125 ppm). The intervention will take place for 6 months.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes of systolic blood pressure | Changes on systolic blood pressure will be measured at the beginning, 3 and 6 months after the intervention. It will be measured in a sitting position and after 5 minutes of rest, using an OMRON HEM 705 CP device, with the cuff adapted to each child, on the left arm. Systolic and diastolic blood pressure will be measured in duplicate, and the mean of both measurements will be recorded, in millimeters of mercury. | 0 month; 3 month; 6 month | 
| Change from Baseline Diastolic Blood Pressure at 6 months | Changes on systolic and diastolic blood pressure will be measured at the beginning, 3 and 6 months after the intervention. It will be measured in a sitting position and after 5 minutes of rest, using an OMRON HEM 705 CP device, with the cuff adapted to each child, on the left arm. Systolic and diastolic blood pressure will be measured in duplicate, and the mean of both measurements will be recorded, in millimeters of mercury. | 0 month; 3 month; 6 month | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| 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. | 0 month; 3 month; 6 month | 
| 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. | 0 month; 3 month; 6 month | 
| 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. | 0 month; 3 month; 6 month | 
| 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. | 0 month; 3 month; 6 month | 
| 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 | 0 month; 3 month; 6 month | 
| 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 | 0 month; 3 month; 6 month | 
| Hemogram | Changes on blood count will be analyzed on an automated analyzer | 0 month; 3 month; 6 month | 
| 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) | 0 month; 3 month; 6 month | 
| Blood insulin | Changes on plasma insulin will be analyzed by radioimmunoassay (RIA) using an automated microparticle analyzer. (AxSYM; Abbott Laboratories, Abbott Park, IL, USA). | 0 month; 3 month; 6 month | 
| Insulin resistance | Changes on insulin resistance (IR) will be calculated by homeostatic model assessment of IR (HOMA-IR). | 0 month; 3 month; 6 month | 
| 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). | 0 month; 3 month; 6 month | 
| 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). | 0 month; 3 month; 6 month | 
| 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. | 0 month; 3 month; 6 month | 
| Antioxidant glutathione reductase activity in red blood cells | Assessing changes of glutathione reductase activity will be carried out using standardized spectrophotometric methods. | 0 month; 3 month; 6 month | 
| Antioxidant glutathione peroxidase activity in red blood cells | Assessing changes of glutathione peroxidase activity will be carried out using standardized spectrophotometric methods. | 0 month; 3 month; 6 month | 
| Antioxidant superoxide dismutase activity in red blood cells | Assessing changes of superoxide dismutase activity will be carried out using standardized spectrophotometric methods. | 0 month; 3 month; 6 month | 
| Antioxidant catalase activity in red blood cells | Assessing changes of catalase activity will be carried out using standardized spectrophotometric methods. | 0 month; 3 month; 6 month | 
| 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 | 0 month; 3 month; 6 month | 
| Urine endothelin-1 | Assessing changes of endothelin-1 will be analyzed in plasma using an ELISA kit. | 0 month; 3 month; 6 month | 
| Analysis of oxLDL in plasma | Assessing changes of oxidized LDL from the baseline to the end of each treatment, by using an ELISA kit | 0 month; 3 month; 6 month | 
| F2-Isoprostanes in urine | Assessing changes of F2-Isoprostanes will be carried out using an ELISA kit. | 0 month; 3 month; 6 month | 
| 8-hydroxy-deoxy-Guanosine in urine | Assessing changes of 8-hydroxy-deoxy-Guanosine will be carried out using an ELISA kit. | 0 month; 3 month; 6 month | 
| 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. | 0 month; 6 month | 
| Plasma metabolomic analysis | A liquid chromatography platform coupled to a spectrophotometer will be used mass (LC/MS) to determine metabolic profiles by targeted analyses | 0 month; 6 month | 
| Urine metabolomic analysis | A liquid chromatography platform coupled to a spectrophotometer will be used mass (LC/MS) to determine metabolic profiles by targeted analyses | 0 month; 3 month; 6 month | 
| 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. | 0 month | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Extra virgin olive oil, Polyphenols, Triterpenic acids, Hipertensive children