Study of the Consumption of a Food Supplement in a Group of Healthy People

Study Overview
=================
Brief Summary
-----------------
Bioactive supplements might display relevant therapeutic properties according to validated molecular effects. Herein, the effect of a supplement based on diterpenes from Rosmarinus Officinalis L. and alkylglycerols with proven properties against signaling pathways involved in tumorigenesis is evaluated. The biological and molecular effects of this supplement, mainly based on expected effects on immune and genetic modulatory properties is investigated. For this purpose, 60 healthy volunteers were enrolled in a six week, double-blind, randomized and parallel pilot study with two study arms -rosemary and alkylglycerol containing capsules and control capsules. The study includes the analysis of (1) immunological parameters (ex vivo cytokine profile of LPS stimulated PBMC and PBMC phenotyping by cluster differentiation (CD) markers), (2) regulation of the expression of genes linked to immuno-modulation, inflammation, oxidative stress response and cancer, and (3) the analysis of correlation of selected genetic variants (SNPs) with the differential responses among individuals.

Official Title
-----------------
Clinical Study to Evaluate the Effectiveness and Safety of the Consumption of a Food Supplement in a Group of Healthy People

Conditions
-----------------
Immune System and Related Disorders

Intervention / Treatment
-----------------
* Dietary Supplement: Dietary supplementation with capsules containing rosemary and alkylglycerol
* Dietary Supplement: Dietary supplementation with control capsules


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: Age between 18 and 55 years Adequate understanding of the study. Willingness to complete the entire treatment. Exclusion Criteria: BMI >30 Diagnosis of type 2 diabetes mellitus (T2D), hypertension, dyslipidemia or other cardiometabolic disorders Impaired cognitive function; Diagnosed hepatic, renal, or cardiovascular disease Subjects with primary immunodeficiency disorders, consumption of drugs with influence on the immune system, splenectomy. Presence of other pathologies like asthma, food allergies, Crohn's, myasthenia gravis, lupus Consumption of vitamins, minerals, supplements of antioxidant extracts or protein supplements in the 2 weeks prior to the start of the study Subjects treated with drugs affecting the lipid or glycemic profile during the previous 30 days Consumption of anticoagulants or antiplatelet agents, cyclosporine, acetylsalicylic acid, antihistamines or sedatives Hypersensitivity to rosemary, to its components or other members of the family of lipped plants or to soybean as excipient of the capsules Allergy or hypersensitivity to fish Habitual smoking or high consumption alcohol Pregnant or lactating women High-intensity physical exercise.

Ages Eligible for Study
-----------------
Minimum Age: 18 Years
Maximum Age: 55 Years

Sexes Eligible for Study
-----------------
All

Accepts Healthy Volunteers
-----------------
Accepts Healthy Volunteers

Study Plan
=================
How is the study designed?
-----------------

Design Details

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


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Rosemary and alkylglycerol capsules<br> | Dietary Supplement: Dietary supplementation with capsules containing rosemary and alkylglycerol<br> <br> |
| Placebo Comparator: Control capsules<br> | Dietary Supplement: Dietary supplementation with control capsules<br> <br> |

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in ex vivo cytokine profile produced by lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMCs) | Isolated PBMCs were first incubated for 12h, and then after LPS treated. Supernatants were recovered to determine concentrations of Interleukin (IL) -1B, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IFNy and TNFa using a magnetic bead-based immunoassay (Human High Sensitivity T Cell Magnetic Bead Panel A MAGPIX-Luminex) kit from Millipore, following the manufacturer's instructions. A minimum of 50 beads per parameter was analyzed by the MAGPIX-Luminex system. Raw data (median ﬂuorescence intensity, MFI) were analyzed with the xPONENT software 4.1. | Baseline and after 6 weeks of treatment | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in oxidative stress status | To evaluate the changes in the oxidative stress the following biomarkers were measured in urine samples: Oxidized-low density lipoproteins measured by sandwich enzyme-linked immunosorbent assay (ELISA) by using the monoclonal antibody mAb-4E6 (Mercodia AB, Sweden) Isoprostanes and thromboxane B2 quantified by competitive ELISA (Enzo Biochem, Inc., NY, USA and Oxford Biomedical Research, MI, USA, respectively). | Baseline and after 6 weeks | 
| Changes in lipid profile | To evaluate lipid improvements the following measurements were considered: Triacylglycerol, Total Cholesterol, low Density Lipoprotein and High-Density Lipoprotein measured by routine laboratory (CQS, Madrid, Spain, which follows the UNE-ISO 15189:2007 directives) methods. | Baseline and after 6 weeks | 
| Gene expression analysis | Gene-expression assays were performed in a HT-7900 Fast Real time PCR. GAPDH was used as endogenous control. RT-StatMiner software (Integromics® Inc., Madison, USA) was used to detect and determine the quality control and differential expression analyses. The Expression Suite Software (Life Technologies) program was used to obtain the Ct data. The ΔCt (Ct gene-CtGAPDH) was calculated and then the relative expression (RQ) between visits was calculated (V3-V1) following the 2-ΔΔCt method (Livak and Schmittgen 2001) | Baseline and after 6 weeks | 
| DNA genotyping | Genotyping was performed using the QuantStudio 12 K Flex Real-Time PCR System (Life Technologies Inc., Carlsbad, CA) with a TaqMan OpenArray plates. Single nucleotide polymorphisms (SNPs) involved in different parts of the pathogenic processes of inflammation, immune system, obesity, lipid metabolism, redox homeostasis and cancer, were analyzed using TaqMan Genotyper software. | Baseline | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
immuno-nutrition, colon cancer, alkylglycerols, Rosmarinus officinalis L