<clinical_study>
  <!-- This xml conforms to an XML Schema at:
    https://clinicaltrials.gov/ct2/html/images/info/public.xsd -->
  <required_header>
    <download_date>ClinicalTrials.gov processed this data on September 20, 2023</download_date>
    <link_text>Link to the current ClinicalTrials.gov record.</link_text>
    <url>https://clinicaltrials.gov/ct2/show/NCT03232099</url>
  </required_header>
  <id_info>
    <org_study_id>57379216.0.0000.0068</org_study_id>
    <secondary_id>2015/212606</secondary_id>
    <nct_id>NCT03232099</nct_id>
  </id_info>
  <brief_title>Red Wine Effects Upon Gut Flora and Plasma Levels of Trimethylamine-N-oxide (TMAO) - WineFlora Study</brief_title>
  <official_title>Evaluation of Red Wine Effects Upon Gut Flora and Plasma Levels of Trimethylamine-N-oxide (TMAO) in Patients With Established Atherosclerotic Disease</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>University of Sao Paulo General Hospital</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>Fundação de Amparo à Pesquisa do Estado de São Paulo</agency>
      <agency_class>Other</agency_class>
    </collaborator>
    <collaborator>
      <agency>IBRAVIN</agency>
      <agency_class>Other</agency_class>
    </collaborator>
  </sponsors>
  <source>University of Sao Paulo General Hospital</source>
  <oversight_info>
    <has_dmc>No</has_dmc>
    <is_fda_regulated_drug>No</is_fda_regulated_drug>
    <is_fda_regulated_device>No</is_fda_regulated_device>
  </oversight_info>
  <brief_summary>
    <textblock>
      Recent evidence indicates that Trimethylamine-N-oxide (TMAO) is a pro-atherosclerotic,&#xD;
      phosphatidylcholine-dependent metabolite of diet and intestinal flora. Food substrates derive&#xD;
      from carnitine and phosphatidylcholine (lecithin), present mainly in eggs, red meat, liver&#xD;
      and pork. The intestinal flora pattern that favors the formation of TMAO is very similar to&#xD;
      that which predisposes to insulin resistance and obesity: a high proportion between phylum&#xD;
      Firmicutes over Bacteroidetes. The intestinal microbiota is sensitive and variable; the use&#xD;
      of prebiotics and probiotics can change the relationship between Firmicutes/Bacteroidetes&#xD;
      phyla. Red wine (RW), for its composition with polyphenols and possible bactericidal role,&#xD;
      may play a role in the intestinal flora modification and could promote proliferation of&#xD;
      beneficial bacteria. However, the influence of RW on TMAO is not known. This is the&#xD;
      hypothesis to be tested in this trial. METHODS: This is a prospective, crossover, randomized,&#xD;
      controlled trial with patients from Heart Institute (InCor), FMUSP and volunteers recruited&#xD;
      through press releases. We will evaluate 42 patients, all men, with established&#xD;
      atherosclerotic disease. Patients will be evaluated in a crossed manner: each subject&#xD;
      receives both treatments, intervention and control (in random order), and they will be&#xD;
      divided into 2 groups: A and B. In the first intervention stage, after 2 weeks of washout for&#xD;
      all patients , group A receives Red Wine (RW) and group B is the control, abstemious. In the&#xD;
      2nd stage of intervention, after 2 weeks of washout for all patients the groups are inverted:&#xD;
      group B receives RW; and group A will be abstemious. In the period with wine intervention,&#xD;
      patients will receive 250 mL/day of red wine per day, for 5 days of the week, for 3 weeks.&#xD;
      Patients will maintain their usual diet without the use of prebiotics or probiotics, or other&#xD;
      polyphenolic derivatives. At the beginning and at the end of each stage, patients will be&#xD;
      submitted to serum TMAO and intestinal microbiota evaluation. For the intestinal microbiota&#xD;
      evaluation, the new generation sequencing will be used in the highly preserved portion of the&#xD;
      16S subunit of the rRNA gene. The determination of TMAO in plasma will be by liquid&#xD;
      chromatography coupled to mass spectrometry. Expected results: It is expected to determine if&#xD;
      RW acts on the intestinal flora to the point of influencing plasma TMAO concentration.&#xD;
    </textblock>
  </brief_summary>
  <overall_status>Completed</overall_status>
  <start_date type="Actual">August 31, 2016</start_date>
  <completion_date type="Actual">April 15, 2018</completion_date>
  <primary_completion_date type="Actual">March 22, 2018</primary_completion_date>
  <phase>N/A</phase>
  <study_type>Interventional</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <allocation>Randomized</allocation>
    <intervention_model>Crossover Assignment</intervention_model>
    <intervention_model_description>Forty two men with established atherosclerotic disease will be studied:after a 2 week washout period, each subject receives both treatments, intervention and control</intervention_model_description>
    <primary_purpose>Basic Science</primary_purpose>
    <masking>None (Open Label)</masking>
  </study_design_info>
  <primary_outcome>
    <measure>Gut flora modification assessed by 16S ribosomal rNA gene sequencing and plasma concentration of TMAO changes measured by liquid chromatography coupled to mass spectrometry in 42 patients after ingestion 250 mL of Red Wine 5 days/week, for 3 weeks</measure>
    <time_frame>up to 10 weeks</time_frame>
    <description>Patients will be divided into 2 groups: A and B. In the first intervention stage, after 2 weeks of washout for all patients , group A receives Red Wine (RW) and group B is the control, abstemious. In the 2nd stage of intervention, after 2 weeks of washout for all patients the groups are inverted: group B receives RW; and group A will be abstemious. In the period with wine intervention, patients will receive 250 mL/day of red wine per day, for 5 days of the week, for 3 weeks. Patients will maintain their usual diet without the use of prebiotics or probiotics, or other polyphenolic derivatives. At the beginning and at the end of each stage, patients will be submitted to serum TMAO and intestinal microbiota evaluation. For the intestinal microbiota evaluation, the new generation sequencing will be used in the highly preserved portion of the 16S subunit of the rRNA gene. The determination of TMAO in plasma will be by liquid chromatography coupled to mass spectrometry</description>
  </primary_outcome>
  <number_of_arms>2</number_of_arms>
  <enrollment type="Actual">42</enrollment>
  <condition>Trimethylamine-N-oxide</condition>
  <condition>Gut Microbiota</condition>
  <condition>Effects of Red Wine</condition>
  <condition>Atherosclerosis</condition>
  <arm_group>
    <arm_group_label>Red Wine group (RW)</arm_group_label>
    <arm_group_type>Experimental</arm_group_type>
    <description>After a two weeks washout period,in the intervening period with red wine, patients will receive 250 mL / day of red wine per Day, 5 days a week, for 3 weeks.</description>
  </arm_group>
  <arm_group>
    <arm_group_label>Abstemious</arm_group_label>
    <arm_group_type>Active Comparator</arm_group_type>
    <description>After a two weeks washout period,in the Abstemious period, patients should not consume any kind of alcohol beverages, for 3 weeks</description>
  </arm_group>
  <intervention>
    <intervention_type>Dietary Supplement</intervention_type>
    <intervention_name>Red Wine consumption</intervention_name>
    <description>After a two weeks washout period,in the intervening period with red wine, patients will receive 250 mL / day of red wine per Day, 5 days a week, for 3 weeks.</description>
    <arm_group_label>Red Wine group (RW)</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Other</intervention_type>
    <intervention_name>Period without alcohol consumption</intervention_name>
    <description>After a two weeks washout period,in the Abstemious period, patients should not consume any kind of alcohol beverages, for 3 weeks</description>
    <arm_group_label>Abstemious</arm_group_label>
    <other_name>Abstain Period</other_name>
  </intervention>
  <eligibility>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          1. Men, 45-70 years&#xD;
&#xD;
          2. Established atherosclerotic diseases defined as:&#xD;
&#xD;
        A. Previous history of CAD: angina or acute myocardial infarction (AMI), myocardial&#xD;
        revascularization or angioplasty, angiographic evidence of stenosis ≥50% in at least one of&#xD;
        the major coronary arteries B. Cerebrovascular disease: transient ischemic stroke or&#xD;
        cerebrovascular accident C. Peripheral arterial disease: clinical evidence of extracoronary&#xD;
        atherosclerosis&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          1. Acute events in the last 30 days (AMI, troponin elevation, coronary angioplasty or&#xD;
             coronary artery bypass grafting)&#xD;
&#xD;
          2. Heart Failure (NYHA functional class ≥ II)&#xD;
&#xD;
          3. Renal Failure (clearance creatinine &lt;30 mL / min by the Cockcroft-Gault formula)&#xD;
&#xD;
          4. Hepatic Failure&#xD;
&#xD;
          5. Gastro-intestinal cancer&#xD;
&#xD;
          6. Intestinal inflammatory diseases&#xD;
&#xD;
          7. Obstructive biliary diseases&#xD;
&#xD;
          8. Prior gastrointestinal surgeries: cholecystectomy or colectomy&#xD;
&#xD;
          9. Use of antibiotics within the last 2 months or during protocol&#xD;
&#xD;
         10. Alcoholism or alcohol intolerance&#xD;
&#xD;
         11. Diabetes mellitus or use of antidiabetics drugs&#xD;
      </textblock>
    </criteria>
    <gender>Male</gender>
    <gender_based>Yes</gender_based>
    <gender_description>Male</gender_description>
    <minimum_age>45 Years</minimum_age>
    <maximum_age>70 Years</maximum_age>
    <healthy_volunteers>No</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Protasio L da Luz, Professor</last_name>
    <role>Study Chair</role>
    <affiliation>Instituto do Coração - InCor - University of Sao Paulo General Hospital</affiliation>
  </overall_official>
  <overall_official>
    <last_name>Elisa A Haas, MD</last_name>
    <role>Principal Investigator</role>
    <affiliation>Instituto do Coração - InCor - University of Sao Paulo General Hospital</affiliation>
  </overall_official>
  <reference>
    <citation>Tang WH, Wang Z, Levison BS, Koeth RA, Britt EB, Fu X, Wu Y, Hazen SL. Intestinal microbial metabolism of phosphatidylcholine and cardiovascular risk. N Engl J Med. 2013 Apr 25;368(17):1575-84. doi: 10.1056/NEJMoa1109400.</citation>
    <PMID>23614584</PMID>
  </reference>
  <reference>
    <citation>Zoetendal EG, Collier CT, Koike S, Mackie RI, Gaskins HR. Molecular ecological analysis of the gastrointestinal microbiota: a review. J Nutr. 2004 Feb;134(2):465-72. doi: 10.1093/jn/134.2.465.</citation>
    <PMID>14747690</PMID>
  </reference>
  <reference>
    <citation>Eckburg PB, Bik EM, Bernstein CN, Purdom E, Dethlefsen L, Sargent M, Gill SR, Nelson KE, Relman DA. Diversity of the human intestinal microbial flora. Science. 2005 Jun 10;308(5728):1635-8. doi: 10.1126/science.1110591. Epub 2005 Apr 14.</citation>
    <PMID>15831718</PMID>
  </reference>
  <reference>
    <citation>Andersson AF, Lindberg M, Jakobsson H, Backhed F, Nyren P, Engstrand L. Comparative analysis of human gut microbiota by barcoded pyrosequencing. PLoS One. 2008 Jul 30;3(7):e2836. doi: 10.1371/journal.pone.0002836.</citation>
    <PMID>18665274</PMID>
  </reference>
  <reference>
    <citation>Brown JM, Hazen SL. The gut microbial endocrine organ: bacterially derived signals driving cardiometabolic diseases. Annu Rev Med. 2015;66:343-59. doi: 10.1146/annurev-med-060513-093205.</citation>
    <PMID>25587655</PMID>
  </reference>
  <reference>
    <citation>Ridaura VK, Faith JJ, Rey FE, Cheng J, Duncan AE, Kau AL, Griffin NW, Lombard V, Henrissat B, Bain JR, Muehlbauer MJ, Ilkayeva O, Semenkovich CF, Funai K, Hayashi DK, Lyle BJ, Martini MC, Ursell LK, Clemente JC, Van Treuren W, Walters WA, Knight R, Newgard CB, Heath AC, Gordon JI. Gut microbiota from twins discordant for obesity modulate metabolism in mice. Science. 2013 Sep 6;341(6150):1241214. doi: 10.1126/science.1241214.</citation>
    <PMID>24009397</PMID>
  </reference>
  <reference>
    <citation>Caricilli AM, Saad MJ. The role of gut microbiota on insulin resistance. Nutrients. 2013 Mar 12;5(3):829-51. doi: 10.3390/nu5030829.</citation>
    <PMID>23482058</PMID>
  </reference>
  <verification_date>May 2017</verification_date>
  <study_first_submitted>June 20, 2017</study_first_submitted>
  <study_first_submitted_qc>July 26, 2017</study_first_submitted_qc>
  <study_first_posted type="Actual">July 27, 2017</study_first_posted>
  <last_update_submitted>March 10, 2021</last_update_submitted>
  <last_update_submitted_qc>March 10, 2021</last_update_submitted_qc>
  <last_update_posted type="Actual">March 15, 2021</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor</responsible_party_type>
  </responsible_party>
  <keyword>trimethylamine-N-oxide</keyword>
  <keyword>TMAO</keyword>
  <keyword>Gut Microbiota</keyword>
  <keyword>Gut Flora</keyword>
  <keyword>Atherosclerosis</keyword>
  <keyword>Wine</keyword>
  <keyword>Red Wine</keyword>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Atherosclerosis</mesh_term>
  </condition_browse>
  <patient_data>
    <sharing_ipd>Undecided</sharing_ipd>
  </patient_data>
  <!-- Results have not yet been posted for this study                                          -->
</clinical_study>


download date: ClinicalTrials.gov processed this data on September 20, 2023
link text: Link to the current ClinicalTrials.gov record.
url: https://clinicaltrials.gov/ct2/show/NCT03232099
org study id: 57379216.0.0000.0068
secondary id: 2015/212606
nct id: NCT03232099
lead sponsor: 
collaborator: 
collaborator: 
has dmc: No
is fda regulated drug: No
is fda regulated device: No
Recent evidence indicates that Trimethylamine-N-oxide (TMAO) is a pro-atherosclerotic,
      phosphatidylcholine-dependent metabolite of diet and intestinal flora. Food substrates derive
      from carnitine and phosphatidylcholine (lecithin), present mainly in eggs, red meat, liver
      and pork. The intestinal flora pattern that favors the formation of TMAO is very similar to
      that which predisposes to insulin resistance and obesity: a high proportion between phylum
      Firmicutes over Bacteroidetes. The intestinal microbiota is sensitive and variable; the use
      of prebiotics and probiotics can change the relationship between Firmicutes/Bacteroidetes
      phyla. Red wine (RW), for its composition with polyphenols and possible bactericidal role,
      may play a role in the intestinal flora modification and could promote proliferation of
      beneficial bacteria. However, the influence of RW on TMAO is not known. This is the
      hypothesis to be tested in this trial. METHODS: This is a prospective, crossover, randomized,
      controlled trial with patients from Heart Institute (InCor), FMUSP and volunteers recruited
      through press releases. We will evaluate 42 patients, all men, with established
      atherosclerotic disease. Patients will be evaluated in a crossed manner: each subject
      receives both treatments, intervention and control (in random order), and they will be
      divided into 2 groups: A and B. In the first intervention stage, after 2 weeks of washout for
      all patients , group A receives Red Wine (RW) and group B is the control, abstemious. In the
      2nd stage of intervention, after 2 weeks of washout for all patients the groups are inverted:
      group B receives RW; and group A will be abstemious. In the period with wine intervention,
      patients will receive 250 mL/day of red wine per day, for 5 days of the week, for 3 weeks.
      Patients will maintain their usual diet without the use of prebiotics or probiotics, or other
      polyphenolic derivatives. At the beginning and at the end of each stage, patients will be
      submitted to serum TMAO and intestinal microbiota evaluation. For the intestinal microbiota
      evaluation, the new generation sequencing will be used in the highly preserved portion of the
      16S subunit of the rRNA gene. The determination of TMAO in plasma will be by liquid
      chromatography coupled to mass spectrometry. Expected results: It is expected to determine if
      RW acts on the intestinal flora to the point of influencing plasma TMAO concentration.
allocation: Randomized
intervention model: Crossover Assignment
intervention model description: Forty two men with established atherosclerotic disease will be studied:after a 2 week washout period, each subject receives both treatments, intervention and control
primary purpose: Basic Science
masking: None (Open Label)
measure: Gut flora modification assessed by 16S ribosomal rNA gene sequencing and plasma concentration of TMAO changes measured by liquid chromatography coupled to mass spectrometry in 42 patients after ingestion 250 mL of Red Wine 5 days/week, for 3 weeks
time frame: up to 10 weeks
description: Patients will be divided into 2 groups: A and B. In the first intervention stage, after 2 weeks of washout for all patients , group A receives Red Wine (RW) and group B is the control, abstemious. In the 2nd stage of intervention, after 2 weeks of washout for all patients the groups are inverted: group B receives RW; and group A will be abstemious. In the period with wine intervention, patients will receive 250 mL/day of red wine per day, for 5 days of the week, for 3 weeks. Patients will maintain their usual diet without the use of prebiotics or probiotics, or other polyphenolic derivatives. At the beginning and at the end of each stage, patients will be submitted to serum TMAO and intestinal microbiota evaluation. For the intestinal microbiota evaluation, the new generation sequencing will be used in the highly preserved portion of the 16S subunit of the rRNA gene. The determination of TMAO in plasma will be by liquid chromatography coupled to mass spectrometry
arm group label: Red Wine group (RW)
arm group type: Experimental
description: After a two weeks washout period,in the intervening period with red wine, patients will receive 250 mL / day of red wine per Day, 5 days a week, for 3 weeks.
arm group label: Abstemious
arm group type: Active Comparator
description: After a two weeks washout period,in the Abstemious period, patients should not consume any kind of alcohol beverages, for 3 weeks
intervention type: Dietary Supplement
intervention name: Red Wine consumption
description: After a two weeks washout period,in the intervening period with red wine, patients will receive 250 mL / day of red wine per Day, 5 days a week, for 3 weeks.
arm group label: Red Wine group (RW)
intervention type: Other
intervention name: Period without alcohol consumption
description: After a two weeks washout period,in the Abstemious period, patients should not consume any kind of alcohol beverages, for 3 weeks
arm group label: Abstemious
other name: Abstain Period
criteria: 
gender: Male
gender based: Yes
gender description: Male
minimum age: 45 Years
maximum age: 70 Years
healthy volunteers: No
last name: Protasio L da Luz, Professor
role: Study Chair
affiliation: Instituto do Coração - InCor - University of Sao Paulo General Hospital
last name: Elisa A Haas, MD
role: Principal Investigator
affiliation: Instituto do Coração - InCor - University of Sao Paulo General Hospital
citation: Tang WH, Wang Z, Levison BS, Koeth RA, Britt EB, Fu X, Wu Y, Hazen SL. Intestinal microbial metabolism of phosphatidylcholine and cardiovascular risk. N Engl J Med. 2013 Apr 25;368(17):1575-84. doi: 10.1056/NEJMoa1109400.
PMID: 23614584
citation: Zoetendal EG, Collier CT, Koike S, Mackie RI, Gaskins HR. Molecular ecological analysis of the gastrointestinal microbiota: a review. J Nutr. 2004 Feb;134(2):465-72. doi: 10.1093/jn/134.2.465.
PMID: 14747690
citation: Eckburg PB, Bik EM, Bernstein CN, Purdom E, Dethlefsen L, Sargent M, Gill SR, Nelson KE, Relman DA. Diversity of the human intestinal microbial flora. Science. 2005 Jun 10;308(5728):1635-8. doi: 10.1126/science.1110591. Epub 2005 Apr 14.
PMID: 15831718
citation: Andersson AF, Lindberg M, Jakobsson H, Backhed F, Nyren P, Engstrand L. Comparative analysis of human gut microbiota by barcoded pyrosequencing. PLoS One. 2008 Jul 30;3(7):e2836. doi: 10.1371/journal.pone.0002836.
PMID: 18665274
citation: Brown JM, Hazen SL. The gut microbial endocrine organ: bacterially derived signals driving cardiometabolic diseases. Annu Rev Med. 2015;66:343-59. doi: 10.1146/annurev-med-060513-093205.
PMID: 25587655
citation: Ridaura VK, Faith JJ, Rey FE, Cheng J, Duncan AE, Kau AL, Griffin NW, Lombard V, Henrissat B, Bain JR, Muehlbauer MJ, Ilkayeva O, Semenkovich CF, Funai K, Hayashi DK, Lyle BJ, Martini MC, Ursell LK, Clemente JC, Van Treuren W, Walters WA, Knight R, Newgard CB, Heath AC, Gordon JI. Gut microbiota from twins discordant for obesity modulate metabolism in mice. Science. 2013 Sep 6;341(6150):1241214. doi: 10.1126/science.1241214.
PMID: 24009397
citation: Caricilli AM, Saad MJ. The role of gut microbiota on insulin resistance. Nutrients. 2013 Mar 12;5(3):829-51. doi: 10.3390/nu5030829.
PMID: 23482058
responsible party type: Sponsor
mesh term: Atherosclerosis
sharing ipd: Undecided