- ICH GCP
- US Clinical Trials Registry
- Clinical Trial NCT04208451
The Effects of One Month Consumption of Standardized Aronia Melanocarpa Extract on Anemia in Patients on Hemodialysis
The Effects of One Month Consumption of Standardized Aronia Melanocarpa Extract on Anemia in Patients on Hemodialysis: Focus on Oxidative Stress and Inflammation
Study Overview
Status
Conditions
Intervention / Treatment
Detailed Description
- Patients The study included 30 patients with chronic kidney failure on dialysis treatment at the Center for Nephrology and Dialysis of the Clinical Center Kragujevac. Inclusion criteria were: regular dialysis treatment for more than 3 months, 3 times a week and hemoglobin values lower than 110g / L. Exclusion criteria were: hemoglobin values lower than 80g/L, the use of antioxidant and immunosuppressive therapy, uncontrolled malignancies, proven active bleeding and presence of systemic inflammation or active infection.
- Ethical approval The research was conducted with respect to the Helsinki Declaration on Medical Research, the approval of the Ethics Committee of the Clinical Center Kragujevac No 01-14-3039. All subjects provided written informed consent before study enrollment.
- Used plant extract Standardized Aronia extract (SAE) is official product of pharmaceutical company Pharmanova (Belgrade, Serbia); nevertheless, procedure of extraction was done by EU-Chem company (Belgrade, Serbia). This product contains 400mg/30ml of polyphenols, while the recommended daily dosage is 30ml.
- Blood sampling Patients included in the study consumed standardized Aronia melanocarpa extract (30ml/day) for 30 days. On the day 0, before SAE consumption, blood samples from all patients were collected for hematologic analyzes, inflammation parameters, oxidative stress parameters and antioxidant protection before the dialysis procedure. All mentioned parameters were determined again after consuming the product, ie. on the 30th day of the study.
- Analytic Procedures Venous blood samples (2 x 4.5ml) were taken before the start of supplementation (zero day) and at the end of supplementation (30th day). Vacuum tubes with sodium citrate were used for blood sampling. The first sample was used for routine hematologic analyzes, the second sample was used for plasma and erythrocyte lysate extraction in order to determinate redox and inflammatory status.
Evaluation of Systemic Redox State Plasma samples were used for determination of the levels of the following pro-oxidants: superoxide anion radical (O2-), hydrogen peroxide (H2O2), nitrites (NO2-) and index of lipid peroxidation measured as thiobarbituric acid reactive substances (TBARS), while the parameters of antioxidative defence system, such as activities of superoxide dismutase (SOD) and catalase (CAT) and level of reduced glutathione (GSH) were determined in erythrocytes lysates samples.
A) Index of lipid peroxidation (TBARS) determination The degree of lipid peroxidation in the plasma samples was estimated by measuring TBARS, using 1% thiobarbituric acid in 0.05 NaOH, which was incubated with the sample at 100°C for 15 min and measured at 530 nm. TBA extract was obtained by combining 0.8 ml sample and 0.4 ml trichloro acetic acid (TCA); afterwards, the samples were put on ice for 10 min and centrifuged for 15 min at 6000 rpm. (1).
b) Nitrite determination (NO2-)
Nitric oxide (NO) decomposes rapidly to form stable nitrite/nitrate products. The NO2- level was measured and used as an index of NO production, using Griess's reagent. For NO2 - determination in plasma 0.1 ml 3 N PCA (perchloride acid), 0.4 ml 20 mM ethylenediaminetetraacetic acid (EDTA) and 0.2 ml plasma were put on ice for 15 min, then centrifuged for 15 min at 6000 rpm. After pouring off the supernatant, 220 μl K2CO3 was added. Nitrites were measured at 550 nm. (2)
Superoxide anion radical determination (O2-)
Superoxide anion radical concentrations in plasma samples were measured using the NTB (Nitro Blue Tetrazolium) reagent in TRIS buffer (assay mixture). The measurement was performed at a wavelength of 530 nm. (3).
Hydrogen peroxide determination (H2O2)
The measurement of H2O2 was based on the oxidation of phenol red by H2O2 in a reaction catalyzed by horseradish peroxidase. Two hundred microlitres of perfusate or plasma was precipitated using 800 mL of freshly prepared phenol red solution; 10 μL of (1:20) horseradish peroxidase (made immediately before use) was subsequently added. The level of H2O2 was measured at 610 nm (4).
Determination of reduced glutathione (GSH)
The level of reduced glutathione (GSH) was determined based on GSH oxidation via 5,5-dithiobis-6,2-nitrobenzoic acid. GSH extract was obtained by combining 0.1 ml 0.1% EDTA, 400 μl hemolysate, and 750 μl precipitation solution (containing 1.67 g metaphosphoric acid, 0.2 g EDTA, 30 g NaCl, and filled with distilled water until 100 ml; the solution is stable for 3 weeks at +4C°). The level of GSH was measured at 420 nm (5).
- Determination of antioxidant enzymes (SOD, CAT)
Isolated RBCs were washed three times with three volumes of ice-cold 0.9 mmol/l NaCl, and hemolysates containing about 50 g Hb/l were used for the determination of CAT activity. CAT buffer, prepared hemolysate sample, and 10 mM H2O2 were used for CAT determination. Detection was performed at 360 nm. SOD activity was determined by the epinephrine method. Hemolysate was mixed with carbonate buffer, and then epinephrine was added. Detection was performed at 470 nm (6-10).
Evaluation of the inflammatory status In order to evaluate the inflammatory status of patients, following parameters were measured at both points of interest (0 and 30th day): serum C-reactive protein concentration - CRP and tumor necrosis factor concentration - TNF-α.
The serum C- reactive protein (CRP) concentration was determined by the turbidimetric method on the Olympus AU680. The normal serum CRP concentration is ≤ 5 mg/L. Microinflammation is defined as the concentration of CRP in the serum of 5 mg/L.
Plasma TNF-α concentrations were determined by enzyme linked immunoadsorbent assay (ELISA) using commercially available high sensitivity indirect sandwich enzyme-linked immunosorbent assay (Sigma Aldrich).
Evaluation of the hematological parameters
At both points of interest, the values of the following haematological parameters in all patients were analyzed:
Erythrocytes (Er), hemoglobin (Hb), hematocrit (Hct), erythrocyte index- mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), serum lactate dehydrogenase concentration (LDH), serum haptoglobin concentration and iron status: serum iron concentration, ferritin; total iron binding capacity (TIBC), unsaturated iron binding capacity (UIBC), transferrin saturation (TSAT), haptoglobin.
- Statistical Analysis IBM SPSS Statistics 20.0 Desktop for Windows was used for statistical analysis. Shapiro-Wilk test was used to check the distribution of data. Statistical comparisons were performed using the one-way analysis of variance (ANOVA) tests with a Tukey's post hoc test for multiple comparisons, in the case of normal distribution of data between groups, while Kruskal-Wallis was used for comparison between groups where the distribution of data was different than normal. Values of p < 0.05 were considered to be statistically significant.
Study Type
Enrollment (Actual)
Phase
- Not Applicable
Contacts and Locations
Study Locations
-
-
-
Kragujevac, Serbia, 34000
- Faculty of Medical Science
-
-
Participation Criteria
Eligibility Criteria
Ages Eligible for Study
Accepts Healthy Volunteers
Genders Eligible for Study
Description
Inclusion Criteria:
- regular dialysis treatment for more than 3 months, 3 times a week
- hemoglobin values lower than 110g / L
Exclusion Criteria:
- hemoglobin values lower than 80g/L,
- the use of antioxidant and immunosuppressive therapy,
- uncontrolled malignancies,
- proven active bleeding
- presence of systemic inflammation or active infection
Study Plan
How is the study designed?
Design Details
- Primary Purpose: Supportive Care
- Allocation: N/A
- Interventional Model: Single Group Assignment
- Masking: None (Open Label)
Arms and Interventions
Participant Group / Arm |
Intervention / Treatment |
---|---|
Experimental: Experimental group
Patients on hemodialysis who consumed one month Standardized Aronia Melanocrpa extract
|
During 30 days all patients take every day 30ml of Alixir 400 protect
|
What is the study measuring?
Primary Outcome Measures
Outcome Measure |
Measure Description |
Time Frame |
---|---|---|
Anemia control
Time Frame: two months
|
The iron levels Unit µmol/l
|
two months
|
Anemia control
Time Frame: two months
|
The hemoglobin levels Unit g/l
|
two months
|
Anemia control
Time Frame: two months
|
The ferritin levels Unit ng/ml
|
two months
|
Anemia control
Time Frame: two months
|
The transferrin levels Unit g/l
|
two months
|
Secondary Outcome Measures
Outcome Measure |
Measure Description |
Time Frame |
---|---|---|
Oxidative stress
Time Frame: two months
|
Superoxide anion radical, Hydrogen peroxide, Nitrites Units nmol/ml
|
two months
|
Oxidative stress
Time Frame: two months
|
Superoxide dismutase, Catalase Units U/g Hb x 1000
|
two months
|
Oxidative stress
Time Frame: two months
|
Reduced glutathione Unit nmol/l RBC x 1000
|
two months
|
Other Outcome Measures
Outcome Measure |
Measure Description |
Time Frame |
---|---|---|
Inflammation
Time Frame: two months
|
The parameters of inflammation CRP Unit mg/l
|
two months
|
Inflammation
Time Frame: two months
|
The parameters of inflammation TNF alpha Unit pg/ml
|
two months
|
Inflammation
Time Frame: two months
|
The parameters of inflammation white blood cells Unit x 1000000000 cells/l
|
two months
|
Collaborators and Investigators
Sponsor
Collaborators
Investigators
- Principal Investigator: Isidora Milosavljevic, Department of pharmacy, University of Kragujevac
Publications and helpful links
Study record dates
Study Major Dates
Study Start (Actual)
Primary Completion (Actual)
Study Completion (Actual)
Study Registration Dates
First Submitted
First Submitted That Met QC Criteria
First Posted (Actual)
Study Record Updates
Last Update Posted (Actual)
Last Update Submitted That Met QC Criteria
Last Verified
More Information
Terms related to this study
Keywords
Additional Relevant MeSH Terms
Other Study ID Numbers
- Alixir400protect
Plan for Individual participant data (IPD)
Plan to Share Individual Participant Data (IPD)?
Drug and device information, study documents
Studies a U.S. FDA-regulated drug product
Studies a U.S. FDA-regulated device product
This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.
Clinical Trials on Anemia
-
SanofiActive, not recruitingWarm Autoimmune Hemolytic Anemia (wAIHA)United States, Austria, China, Denmark, Germany, Hungary, Italy, Spain, United Kingdom
-
SanofiTerminatedWarm Autoimmune Hemolytic Anemia (wAIHA)United Kingdom, Belgium, Netherlands, France, United States, Germany, Hungary, Italy
-
Hospital Universitario Dr. Jose E. GonzalezCompletedPernicious Anemia | Megaloblastic Anemia NosMexico
-
Assistance Publique - Hôpitaux de ParisNot yet recruitingSevere Aplastic Anemia | Idiopathic Aplastic Anemia | Moderate Aplastic Anemia Requiring Transfusions
-
Abdelwahed, Mai Mahmoud Mohamed, M.D.UnknownAnemia During PregnancyEgypt
-
University of California, DavisInstituto Mexicano del Seguro Social; Thrasher Research Fund; Mexican National... and other collaboratorsCompleted
-
Incyte CorporationActive, not recruitingWarm Autoimmune Hemolytic Anemia (wAIHA)Spain, United States, Austria, Belgium, Canada, France, Germany, Israel, Italy, Japan, Netherlands, Poland, United Kingdom
-
Peking Union Medical College HospitalRecruiting
-
Alexion PharmaceuticalsWithdrawnWarm Autoimmune Hemolytic AnemiaUnited States
Clinical Trials on Alixir 400 Protect
-
Lungpacer Medical Inc.CompletedAcute Respiratory Distress Syndrome | Ventilation Therapy; ComplicationsFrance
-
Weill Medical College of Cornell UniversityNational Institute of Mental Health (NIMH); New York City Department for the...RecruitingDepression | Elder AbuseUnited States
-
Weill Medical College of Cornell UniversityThe Fan Fox & Leslie R. Samuels Foundation, Inc.; New York City Department... and other collaboratorsCompleted
-
Pädagogische Hochschule HeidelbergHeidelberg UniversityUnknown
-
Lund UniversityTerminated
-
LiveLeaf Inc.UnknownAntibiotic-associated Diarrhea
-
Lund UniversityActive, not recruitingAthletic Injuries | Health BehaviorSweden
-
University of MiamiEunice Kennedy Shriver National Institute of Child Health and Human Development... and other collaboratorsCompletedPregnancy | HIVSouth Africa
-
Pädagogische Hochschule HeidelbergDietmar Hopp StiftungUnknownMental Disorders