Beneficial Impact of Orange Juice Consumption on Risk Factors Associated With Cardiovascular Diseases

Study Overview
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Brief Summary
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The primary aim is to examine both the acute and chronic effects of hesperidin consumption from 100% Florida orange juice in various doses on functional and systemic markers associated with cardiovascular disease (CVD) risks.

Detailed Description
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The aim of this study is to compare the effect of different doses of hesperidin in 100% Florida orange juice (OJ) when regularly or postprandially consumed on cardiovascular risk markers; in addition, the plausible role and mechanism of the hesperidin will be investigated. The sample size was calculated using a previously available bibliography using systolic blood pressure (SBP) as the primary outcome measure. A total of 84 subjects per study product group were needed, assuming variance components of approximately 20.0, to detect differences between the three groups (control, orange juice and hesperidin-enriched orange juice (10 mm Hg)) with a bilateral significance level of α=0.05 and a power of 80 %. The sample size was computed to be sufficient to detect differences between treatment groups regarding the evolution in time of SBP levels. Justification of chosen sample size is based on the clinically meaningful difference assigned to δ=10.0 mm Hg, which is equivalent to a difference of approximately 7.4 % in patients with baseline SBP levels of approximately 135 mm Hg. Thus, a sample of 252 participants can be used for the chronic three arm parallel trial design (84 subjects/arm) and will allow us to detect small but clinically relevant differences between the three groups with statistical robustness and direct interpretation in terms of the chronic treatment effect. To the acute postprandial tests, the investigators have chosen n=20 subjects per arm according to the most studies that have addressed the metabolic effects of a postprandial intervention have been performed using a very similar number of subjects with statistically good quality results. The statistical analysis will follow the principles specified in the guidelines of the ICHE9 and CPMP/EWP/908/99 ICHE9 Points to Consider on Multiplicity Issues in Clinical Trials. The continuous efficacy variables will be analyzed by an ANCOVA (analysis of covariance) with the baseline value as a covariate. The efficacy outcomes will be determined using the absolute values and absolute differences from the baseline. The efficacy analysis will be performed using the Available Data Only approach. In addition, the analysis of the primary efficacy variable will be performed with the Baseline Observation Carried Forward approach. A suitable hypothesis test will be applied to the rest of the variables according to the nature of each variable, such as the Fisher exact test for categorical variables, Student's T-test for continuous variables and Mann-Whitney U test for ordinal scale variables. The statistical tests will be applied with an α=0.05 two-sided significance level. Post-hoc analyses and comparisons between pairs of groups will be done as for exploratory purposes. In addition, the statistical plan will be transferred to the application form of the electronic data collection report (e-CDR), which allows the improvement of data management, diminishes human errors (according threshold values of each outcome) and, overall, guarantees the maximum exploitation of human data in the context of statistical analysis.

Official Title
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Randomized, Parallel and Double Blind Placebo-controlled Study for the Evaluation of Both Acute and Chronic Role of Hesperidin Consumption in 100% Orange Juice

Conditions
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Cardiovascular Risk Factors

Intervention / Treatment
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* Dietary Supplement: Control
* Dietary Supplement: 100% Florida OJ
* Dietary Supplement: 100% Florida OJ-enriched


Participation Criteria
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Eligibility Criteria
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Inclusion criteria Men or women 18-65 years old No evidence of chronic disease No familial CVD history Written informed consent provided before the initial screening visit. Blood pressure (with no drug intervention) >120 mm Hg systolic blood pressure ≤ 159 mmHg Exclusion criteria Body mass index (BMI) ≥ 35 kg/m2 Glucose >125 mg/dl Systolic blood pressure ≥ 160 mm Hg and diastolic blood pressure >100 mm Hg or taking antihypertensive medications Total cholesterol >240 mg/dl LDL-cholesterol >160 md/dl TAG >350 Smoking Pregnant or intending to become pregnant Use of medications, antioxidants, or vitamin supplements Chronic alcoholism Intense physical activity (5h/week) Intestinal disorders Following of a vegetarian diet Anemia (hemoglobin ≤13 g/dL in men and ≤12 g/dL in women) Current or past participation in a clinical trial or consumption of a research product in the 30 days prior to inclusion in the study Failure to follow the study guidelines.

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 65 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Prevention
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Quadruple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: Control<br>Control drink (placebo) | Dietary Supplement: Control<br>* 500 mL (250 mL; 2 times/day) of placebo drink for 12 weeks<br>|
| Experimental: Natural Florida orange juice<br>100% Florida orange juice (OJ) (natural content of hesperidin) | Dietary Supplement: 100% Florida OJ<br>* 500 mL (250 mL; 2 times/day) of 100% Florida OJ for 12 weeks<br>|
| Experimental: Enriched Florida orange juice<br>100% Florida orange juice (OJ) (enriched hesperidin content) | Dietary Supplement: 100% Florida OJ-enriched<br>* 500 mL (250 mL; 2 times/day) of 100% Florida OJ-enriched for 12 weeks<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Systolic Blood Pressure | During each visit, SBP will be measured after 5 min in a seated position in a comfortable room by the physician. The measurement will be taken in duplicate at 1-min intervals using an automatic sphygmomanometer (OMRON HEM-907; Peroxfarma, Barcelona, Spain), and the average of the two measurements will be calculated. | Every 2 weeks for a total of 12 weeks. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Diastolic Blood Pressure | During each visit, diastolic BP will be measured after 5 min in a seated position in a comfortable room by the physician. The measurement will be taken in duplicate at 1-min intervals using an automatic sphygmomanometer (OMRON HEM-907; Peroxfarma, Barcelona, Spain), and the average of the two measurements will be calculated. | Every 2 weeks for a total of 12 weeks. | 
| Ischemic reactive hyperemia (IRH) | The endothelial-dependent vasomotor functions will be measured as IRH by a Laser-Doppler linear Periflux 5000 flowmeter (Perimed AB, Järfälla, Stockholm, Sweden) | Every 4 weeks for a total of 12 weeks | 
| Platelet aggregation | Multiplate analyzer, Roche | Every 4 weeks for a total of 12 weeks | 
| Homocysteine | Homocysteine concentrations will be measured by liquid chromatography-mass spectrometry (LC-MS/MS) | Every 4 weeks for a total of 12 weeks | 
| C-reactive protein (inflammatory marker) | High sensitivity C-reactive protein (hsCRP) by standardized methods in a Cobas Mira Plus autoanalyzer (Roche Diagnostics Systems, Madrid, Spain) | Every 4 weeks for a total of 12 weeks | 
| oxidized LDL (as oxidative stress biomarker) | Mercodia Oxidized LDL ELISA kit will be used to measure the oxidized LDL (mU/L). | Every 4 weeks for a total of 12 weeks | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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biomarkers, Cardiovascular disease, transcriptomics, orange juice, hesperidin, systolic blood pressure, metabolomics, cardiovascular risk factor