<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/NCT02209025</url>
  </required_header>
  <id_info>
    <org_study_id>NL46131.068.13/METC 13-3-065</org_study_id>
    <nct_id>NCT02209025</nct_id>
  </id_info>
  <brief_title>Theobromine, Vascular Function and Intestinal apoA-I Production</brief_title>
  <official_title>The Effects of Short-term Theobromine Supplementation on Vascular Function and Intestinal apoA-I Production in Fasting and in the Postprandial State</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>Maastricht University Medical Center</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>Unilever R&amp;D</agency>
      <agency_class>Industry</agency_class>
    </collaborator>
    <collaborator>
      <agency>DSM Nutritional Products, Inc.</agency>
      <agency_class>Industry</agency_class>
    </collaborator>
  </sponsors>
  <source>Maastricht University Medical Center</source>
  <oversight_info>
    <has_dmc>No</has_dmc>
  </oversight_info>
  <brief_summary>
    <textblock>
      Rationale: Despite successful efforts to lower atherogenic serum low-density lipoprotein&#xD;
      (LDL) cholesterol concentrations, a substantial residual cardiovascular risk remains. An&#xD;
      additive strategy to further lower this residual risk may be via raising high-density&#xD;
      lipoprotein (HDL) concentrations, and in particular those of its major protein constituent&#xD;
      apolipoprotein A-I (apoA-I). Based on several studies, theobromine may be a promising&#xD;
      candidate in this respect. Recently, theobromine was shown to increase serum HDL cholesterol&#xD;
      (HDL-C) concentrations by 0.16 mmol/L or 10% and apoA-I levels with 8%. The question is&#xD;
      whether this increase in HDL-C and apoA-I concentrations observed translates into an improved&#xD;
      functionality of the blood vessels. Effects of theobromine on vascular function have never&#xD;
      been evaluated in a placebo controlled human intervention study.&#xD;
&#xD;
      Objective: The primary objective is to evaluate the long-term effects of theobromine on&#xD;
      vascular function in healthy subjects with a slightly lowered HDL-C in the fasting and the&#xD;
      postprandial state. The second primary objective is to evaluate the long-term effects of&#xD;
      theobromine on intestinal apoA-I mRNA and protein expression levels in healthy subjects with&#xD;
      a slightly lowered HDL-C in the fasting and the postprandial state.&#xD;
&#xD;
      Secondary objectives are to study the long-term effects of theobromine on (1) fasting serum&#xD;
      HDL-C concentrations, (2) cholesterol efflux capacity and (3) postprandial lipid and glucose&#xD;
      metabolism.&#xD;
&#xD;
      Study design: A randomized, double-blind, placebo controlled cross-over design. The total&#xD;
      study duration will be 12 weeks, consisting of a 4 week experimental period, a 4 week&#xD;
      wash-out, and a 4 week control period. At the end of the experimental and control periods, a&#xD;
      postprandial test will take place.&#xD;
&#xD;
      Study population: Forty-eight healthy men aged 45-70 years and women aged 50-70 years, with&#xD;
      slightly lowered HDL-C concentrations (men &lt;1.3 mmol/L and women &lt;1.5 mmol/L).&#xD;
&#xD;
      Intervention: During the experimental period, subjects will consume daily at breakfast an&#xD;
      drink containing 500 mg theobromine. During the placebo period, the subjects will consume&#xD;
      daily at breakfast the same drink without theobromine. During the wash-out period, they will&#xD;
      not consume any of the drinks.&#xD;
&#xD;
      Main study parameters/endpoints: Measurements will be performed at the end of both 4-week&#xD;
      intervention periods. The effects of theobromine will be calculated as the absolute&#xD;
      differences between values obtained at the end of each period. The primary endpoint is the&#xD;
      change in vascular function defined as % change in flow-mediated dilation (FMD) after intake&#xD;
      of a daily stressor, a milkshake providing all the three different macronutrients. The second&#xD;
      primary endpoint is the change in apoA-I mRNA and protein expression on the end of each&#xD;
      period 5 hours after intake of the milkshake.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      Rationale: Despite successful efforts to lower atherogenic serum low-density lipoprotein&#xD;
      (LDL) cholesterol concentrations, a substantial residual cardiovascular risk remains. An&#xD;
      additive strategy to further lower this residual risk may be via raising high-density&#xD;
      lipoprotein (HDL) concentrations, and in particular those of its major protein constituent&#xD;
      apolipoprotein A-I (apoA-I). Based on several studies, theobromine may be a promising&#xD;
      candidate in this respect. Recently, theobromine was shown to increase serum HDL cholesterol&#xD;
      (HDL-C) concentrations by 0.16 mmol/L or 10% and apoA-I levels with 8%. The question is&#xD;
      whether this increase in HDL-C and apoA-I concentrations observed translates into an improved&#xD;
      functionality of the blood vessels. Effects of theobromine on vascular function have never&#xD;
      been evaluated in a placebo controlled human intervention study.&#xD;
&#xD;
      Objective: The primary objective is to evaluate the long-term effects of theobromine on&#xD;
      vascular function in healthy subjects with a slightly lowered HDL-C in the fasting and the&#xD;
      postprandial state. The second primary objective is to evaluate the long-term effects of&#xD;
      theobromine on intestinal apoA-I mRNA and protein expression levels in healthy subjects with&#xD;
      a slightly lowered HDL-C in the fasting and the postprandial state.&#xD;
&#xD;
      Secondary objectives are to study the long-term effects of theobromine on (1) fasting serum&#xD;
      HDL-C concentrations, (2) cholesterol efflux capacity and (3) postprandial lipid and glucose&#xD;
      metabolism.&#xD;
&#xD;
      Study design: A randomized, double-blind, placebo controlled cross-over design. The total&#xD;
      study duration will be 12 weeks, consisting of a 4 week experimental period, a 4 week&#xD;
      wash-out, and a 4 week control period. At the end of the experimental and control periods, a&#xD;
      postprandial test will take place.&#xD;
&#xD;
      Study population: Forty-eight healthy men aged 45-70 years and women aged 50-70 years, with&#xD;
      slightly lowered HDL-C concentrations (men &lt;1.3 mmol/L and women &lt;1.5 mmol/L).&#xD;
&#xD;
      Intervention: During the experimental period, subjects will consume daily at breakfast an&#xD;
      drink containing 500 mg theobromine. During the placebo period, the subjects will consume&#xD;
      daily at breakfast the same drink without theobromine. During the wash-out period, they will&#xD;
      not consume any of the drinks.&#xD;
&#xD;
      Main study parameters/endpoints: Measurements will be performed at the end of both 4-week&#xD;
      intervention periods. The effects of theobromine will be calculated as the absolute&#xD;
      differences between values obtained at the end of each period. The primary endpoint is the&#xD;
      change in vascular function defined as % change in flow-mediated dilation (FMD) after intake&#xD;
      of a daily stressor, a milkshake providing all the three different macronutrients. The second&#xD;
      primary endpoint is the change in apoA-I mRNA and protein expression on the end of each&#xD;
      period 5 hours after intake of the milkshake.&#xD;
&#xD;
      Nature and extent of the burden and risks associated with participation, benefit and group&#xD;
      relatedness: Before the study, subjects will be screened twice to determine eligibility&#xD;
      during two visits of 15 minutes. During these visits, body weight, height and blood pressure&#xD;
      will be measured. In addition, a venous blood sample (5.5 mL at each occasion) will be drawn.&#xD;
      During the study, subjects will receive the control and theobromine drinks in random order.&#xD;
      At day 1 and 14 a fasting venous blood sample (16.5 mL and 7 mL respectively) will be drawn&#xD;
      and body weight and blood pressure will be measured. At days 25 and 81 again a fasting blood&#xD;
      sample will be drawn (28 mL). In addition, at day 28 of each experimental period, at t=0&#xD;
      hours, a fasting venous blood sample (30 mL) will be drawn and vascular function will be&#xD;
      measured, followed by a 2.5 h postprandial test with again the vascular function&#xD;
      measurements. As postprandial meal, subjects will receive a high-fat milkshake. Before the&#xD;
      postprandial test a cannula will be placed and blood will be sampled at t=0, 15, 30, 45, 60,&#xD;
      90, 120 and 240 (total volume 129.5 mL). Thus, during the entire 13 (screening and study)&#xD;
      weeks period, in total 444 mL blood will be drawn. Vascular function measurements include a&#xD;
      panel of non-invasive measurements, i.e. FMD, endo peripheral artherial tonometry (endoPAT),&#xD;
      pulse wave velocity (PWV) and retinal imaging. Subjects will be asked to fill out a food&#xD;
      frequency questionnaire two times and to keep a study-diary throughout 12 weeks. On rare&#xD;
      occasions, blood sampling might cause bruises or hematoma. The vascular function measurements&#xD;
      are non-invasive and will cause no burden.&#xD;
&#xD;
      In a subgroup of 10 volunteers, 5 hours after the start of the postprandial test, small&#xD;
      intestinal biopsies will be taken in the duodenum to determine the difference in intestinal&#xD;
      apoA-I mRNA and protein expression. Obtaining small intestinal biopsies by standard flexible&#xD;
      gastroscopy induces local discomfort in the pharynx only during the procedure, which takes&#xD;
      about 15 minutes, and causes a theoretical risk for perforations (1:1000) and an infection&#xD;
      risk of 1:1.800.000, with lower risk for healthy subjects. The complication is serious and&#xD;
      subjects will have to stay in the hospital. If there are serious complaints after the&#xD;
      gastroscopy (e.g. vomiting with or without blood, or stomach ache) subjects are have to warn&#xD;
      the project leader or their physician.&#xD;
&#xD;
      Total time investment for the subjects will be approximately 14.5 hours, and 17.5 hours in&#xD;
      the subgroup from which biopsies will be taken, excluding travel time.&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Completed</overall_status>
  <start_date>June 2014</start_date>
  <completion_date type="Actual">September 2015</completion_date>
  <primary_completion_date type="Actual">September 2015</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>
    <primary_purpose>Prevention</primary_purpose>
    <masking>Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor)</masking>
  </study_design_info>
  <primary_outcome>
    <measure>Flow mediated dilation</measure>
    <time_frame>4 weeks</time_frame>
    <description>FMD measurements after 4 weeks of intake. Fasting and postprandial measurements</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Intestinal apoA-I production</measure>
    <time_frame>After 4 weeks of intake</time_frame>
    <description>Intestinal biopsies after 4 weeks of intake to see changes in apoA-I gene expression</description>
  </primary_outcome>
  <secondary_outcome>
    <measure>HDL cholesterol</measure>
    <time_frame>4 weeks</time_frame>
    <description>HDL serum cholesterol concentrations after 4 weeks of intake</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Vascular measurements</measure>
    <time_frame>After 4 weeks</time_frame>
    <description>Vascular function measurements (PAT, PWV, PWA, retinal images) after 4 weeks of intake.&#xD;
Fasting and postprandial measurements</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>serum lipid metabolism</measure>
    <time_frame>4 weeks</time_frame>
    <description>serum lipid metabolism (total cholesterol, LDL-cholesterol, triglycerides) during fasting and in the postprandial state?</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>glucose metabolism</measure>
    <time_frame>4 weeks</time_frame>
    <description>glucose and insulin concentrations during fasting and in the postprandial state?</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>(apo)lipoprotein metabolism</measure>
    <time_frame>4 weeks</time_frame>
    <description>ApoB concentrations during fasting and in the postprandial state</description>
  </secondary_outcome>
  <number_of_arms>2</number_of_arms>
  <enrollment type="Actual">48</enrollment>
  <condition>Cardiovascular Disease</condition>
  <arm_group>
    <arm_group_label>Placebo drink</arm_group_label>
    <arm_group_type>Placebo Comparator</arm_group_type>
    <description>A drink with the same components as the theobromine drink except for the theobromine</description>
  </arm_group>
  <arm_group>
    <arm_group_label>Theobromine</arm_group_label>
    <arm_group_type>Experimental</arm_group_type>
    <description>500mg theobromine in a drink</description>
  </arm_group>
  <intervention>
    <intervention_type>Dietary Supplement</intervention_type>
    <intervention_name>Theobromine</intervention_name>
    <description>Theobromine a compound for cocoa</description>
    <arm_group_label>Theobromine</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Dietary Supplement</intervention_type>
    <intervention_name>Placebo drink</intervention_name>
    <arm_group_label>Placebo drink</arm_group_label>
  </intervention>
  <eligibility>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  Men aged between 45 and 70 years,&#xD;
&#xD;
          -  Women aged between 50 and 70 years,&#xD;
&#xD;
          -  Serum HDL-C &lt;1.3 mmol/L (men),&#xD;
&#xD;
          -  Serum HDL-C &lt;1.5 mmol/L (women),&#xD;
&#xD;
          -  Serum total cholesterol &lt;8.0 mmol/L,&#xD;
&#xD;
          -  Plasma glucose &lt;7.0 mmol/L,&#xD;
&#xD;
          -  BMI between 25 - 35 kg/m2,&#xD;
&#xD;
          -  Non-smoking,&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  Unstable body weight (weight gain or loss &gt;3 kg in the past 3 months),&#xD;
&#xD;
          -  Any medical condition requiring treatment and/or medication use,&#xD;
&#xD;
          -  Indication for treatment with cholesterol-lowering drugs according to the Dutch&#xD;
             Cholesterol Consensus (30),&#xD;
&#xD;
          -  Use of medication or a medically-prescribed diet known to affect serum lipid or&#xD;
             glucose metabolism. The use of paracetamol is allowed.&#xD;
&#xD;
          -  Active cardiovascular disease (for instance congestive heart failure) or recent (&lt;6&#xD;
             months) event, such as acute myocardial infarction or cerebro-vascular accident.&#xD;
&#xD;
          -  Gastrointestinal diseases (like celiac disease, inflammatory bowel disease, irritable&#xD;
             bowel disease and food allergies) or a history of any gastrointestinal disorders or&#xD;
             complaints.&#xD;
&#xD;
          -  Not willing to stop the consumption of vitamin supplements or fish oil capsules 2&#xD;
             weeks before the start of the study,&#xD;
&#xD;
          -  Men: consumption of &gt;21 glasses of alcohol-containing drinks per week.&#xD;
&#xD;
          -  Women: consumption of &gt;14 glasses of alcohol-containing drinks per week.&#xD;
&#xD;
          -  Abuse of drugs,&#xD;
&#xD;
          -  Extensive exercise,&#xD;
&#xD;
          -  Not willing to abstain from caffeine containing painkillers two weeks prior to the&#xD;
             study and the duration of the study,&#xD;
&#xD;
          -  Not willing to abstain from theobromine rich products from two weeks prior to the&#xD;
             study and the duration of the study:&#xD;
&#xD;
             o Chocolate containing products (see Appendix A).&#xD;
&#xD;
          -  Not willing to abstain from energy drinks from two weeks prior to the study and the&#xD;
             duration of the study, because of the high caffeine content.&#xD;
&#xD;
          -  Not willing to reduce caffeine containing drinks till maximum 4 drinks a day from two&#xD;
             weeks prior to the study and the duration of the study:&#xD;
&#xD;
               -  Coffee (excluding coffee without caffeine) (see Appendix A),&#xD;
&#xD;
               -  Tea (excluding tea without caffeine) (see Appendix A),&#xD;
&#xD;
               -  Cola (see Appendix A).&#xD;
&#xD;
          -  Participation in another biomedical study within 1 month prior to the screening visit,&#xD;
&#xD;
          -  Having donated &gt;150 ml blood within 1 month prior to the screening visit, planning to&#xD;
             donate blood during the study or within one month after finishing the study.&#xD;
&#xD;
          -  Impossible or difficult to puncture as evidenced during the screening visits.&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>45 Years</minimum_age>
    <maximum_age>70 Years</maximum_age>
    <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>J Plat, Prof</last_name>
    <role>Principal Investigator</role>
    <affiliation>Maastricht University Hospital</affiliation>
  </overall_official>
  <location>
    <facility>
      <name>Maastricht University Hospital</name>
      <address>
        <city>Maastricht</city>
        <zip>6200 MD</zip>
        <country>Netherlands</country>
      </address>
    </facility>
  </location>
  <location_countries>
    <country>Netherlands</country>
  </location_countries>
  <verification_date>August 2014</verification_date>
  <study_first_submitted>August 4, 2014</study_first_submitted>
  <study_first_submitted_qc>August 4, 2014</study_first_submitted_qc>
  <study_first_posted type="Estimate">August 5, 2014</study_first_posted>
  <last_update_submitted>September 4, 2015</last_update_submitted>
  <last_update_submitted_qc>September 4, 2015</last_update_submitted_qc>
  <last_update_posted type="Estimate">September 7, 2015</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor</responsible_party_type>
  </responsible_party>
  <keyword>Atherosclerosis</keyword>
  <keyword>Cholesterol</keyword>
  <keyword>Cholesterol, HDL</keyword>
  <keyword>Apolipoprotein A-I</keyword>
  <keyword>Theobromine</keyword>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Cardiovascular Diseases</mesh_term>
  </condition_browse>
  <intervention_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Theobromine</mesh_term>
  </intervention_browse>
  <!-- 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/NCT02209025
org study id: NL46131.068.13/METC 13-3-065
nct id: NCT02209025
lead sponsor: 
collaborator: 
collaborator: 
has dmc: No
Rationale: Despite successful efforts to lower atherogenic serum low-density lipoprotein
      (LDL) cholesterol concentrations, a substantial residual cardiovascular risk remains. An
      additive strategy to further lower this residual risk may be via raising high-density
      lipoprotein (HDL) concentrations, and in particular those of its major protein constituent
      apolipoprotein A-I (apoA-I). Based on several studies, theobromine may be a promising
      candidate in this respect. Recently, theobromine was shown to increase serum HDL cholesterol
      (HDL-C) concentrations by 0.16 mmol/L or 10% and apoA-I levels with 8%. The question is
      whether this increase in HDL-C and apoA-I concentrations observed translates into an improved
      functionality of the blood vessels. Effects of theobromine on vascular function have never
      been evaluated in a placebo controlled human intervention study.

      Objective: The primary objective is to evaluate the long-term effects of theobromine on
      vascular function in healthy subjects with a slightly lowered HDL-C in the fasting and the
      postprandial state. The second primary objective is to evaluate the long-term effects of
      theobromine on intestinal apoA-I mRNA and protein expression levels in healthy subjects with
      a slightly lowered HDL-C in the fasting and the postprandial state.

      Secondary objectives are to study the long-term effects of theobromine on (1) fasting serum
      HDL-C concentrations, (2) cholesterol efflux capacity and (3) postprandial lipid and glucose
      metabolism.

      Study design: A randomized, double-blind, placebo controlled cross-over design. The total
      study duration will be 12 weeks, consisting of a 4 week experimental period, a 4 week
      wash-out, and a 4 week control period. At the end of the experimental and control periods, a
      postprandial test will take place.

      Study population: Forty-eight healthy men aged 45-70 years and women aged 50-70 years, with
      slightly lowered HDL-C concentrations (men <1.3 mmol/L and women <1.5 mmol/L).

      Intervention: During the experimental period, subjects will consume daily at breakfast an
      drink containing 500 mg theobromine. During the placebo period, the subjects will consume
      daily at breakfast the same drink without theobromine. During the wash-out period, they will
      not consume any of the drinks.

      Main study parameters/endpoints: Measurements will be performed at the end of both 4-week
      intervention periods. The effects of theobromine will be calculated as the absolute
      differences between values obtained at the end of each period. The primary endpoint is the
      change in vascular function defined as % change in flow-mediated dilation (FMD) after intake
      of a daily stressor, a milkshake providing all the three different macronutrients. The second
      primary endpoint is the change in apoA-I mRNA and protein expression on the end of each
      period 5 hours after intake of the milkshake.
Rationale: Despite successful efforts to lower atherogenic serum low-density lipoprotein
      (LDL) cholesterol concentrations, a substantial residual cardiovascular risk remains. An
      additive strategy to further lower this residual risk may be via raising high-density
      lipoprotein (HDL) concentrations, and in particular those of its major protein constituent
      apolipoprotein A-I (apoA-I). Based on several studies, theobromine may be a promising
      candidate in this respect. Recently, theobromine was shown to increase serum HDL cholesterol
      (HDL-C) concentrations by 0.16 mmol/L or 10% and apoA-I levels with 8%. The question is
      whether this increase in HDL-C and apoA-I concentrations observed translates into an improved
      functionality of the blood vessels. Effects of theobromine on vascular function have never
      been evaluated in a placebo controlled human intervention study.

      Objective: The primary objective is to evaluate the long-term effects of theobromine on
      vascular function in healthy subjects with a slightly lowered HDL-C in the fasting and the
      postprandial state. The second primary objective is to evaluate the long-term effects of
      theobromine on intestinal apoA-I mRNA and protein expression levels in healthy subjects with
      a slightly lowered HDL-C in the fasting and the postprandial state.

      Secondary objectives are to study the long-term effects of theobromine on (1) fasting serum
      HDL-C concentrations, (2) cholesterol efflux capacity and (3) postprandial lipid and glucose
      metabolism.

      Study design: A randomized, double-blind, placebo controlled cross-over design. The total
      study duration will be 12 weeks, consisting of a 4 week experimental period, a 4 week
      wash-out, and a 4 week control period. At the end of the experimental and control periods, a
      postprandial test will take place.

      Study population: Forty-eight healthy men aged 45-70 years and women aged 50-70 years, with
      slightly lowered HDL-C concentrations (men <1.3 mmol/L and women <1.5 mmol/L).

      Intervention: During the experimental period, subjects will consume daily at breakfast an
      drink containing 500 mg theobromine. During the placebo period, the subjects will consume
      daily at breakfast the same drink without theobromine. During the wash-out period, they will
      not consume any of the drinks.

      Main study parameters/endpoints: Measurements will be performed at the end of both 4-week
      intervention periods. The effects of theobromine will be calculated as the absolute
      differences between values obtained at the end of each period. The primary endpoint is the
      change in vascular function defined as % change in flow-mediated dilation (FMD) after intake
      of a daily stressor, a milkshake providing all the three different macronutrients. The second
      primary endpoint is the change in apoA-I mRNA and protein expression on the end of each
      period 5 hours after intake of the milkshake.

      Nature and extent of the burden and risks associated with participation, benefit and group
      relatedness: Before the study, subjects will be screened twice to determine eligibility
      during two visits of 15 minutes. During these visits, body weight, height and blood pressure
      will be measured. In addition, a venous blood sample (5.5 mL at each occasion) will be drawn.
      During the study, subjects will receive the control and theobromine drinks in random order.
      At day 1 and 14 a fasting venous blood sample (16.5 mL and 7 mL respectively) will be drawn
      and body weight and blood pressure will be measured. At days 25 and 81 again a fasting blood
      sample will be drawn (28 mL). In addition, at day 28 of each experimental period, at t=0
      hours, a fasting venous blood sample (30 mL) will be drawn and vascular function will be
      measured, followed by a 2.5 h postprandial test with again the vascular function
      measurements. As postprandial meal, subjects will receive a high-fat milkshake. Before the
      postprandial test a cannula will be placed and blood will be sampled at t=0, 15, 30, 45, 60,
      90, 120 and 240 (total volume 129.5 mL). Thus, during the entire 13 (screening and study)
      weeks period, in total 444 mL blood will be drawn. Vascular function measurements include a
      panel of non-invasive measurements, i.e. FMD, endo peripheral artherial tonometry (endoPAT),
      pulse wave velocity (PWV) and retinal imaging. Subjects will be asked to fill out a food
      frequency questionnaire two times and to keep a study-diary throughout 12 weeks. On rare
      occasions, blood sampling might cause bruises or hematoma. The vascular function measurements
      are non-invasive and will cause no burden.

      In a subgroup of 10 volunteers, 5 hours after the start of the postprandial test, small
      intestinal biopsies will be taken in the duodenum to determine the difference in intestinal
      apoA-I mRNA and protein expression. Obtaining small intestinal biopsies by standard flexible
      gastroscopy induces local discomfort in the pharynx only during the procedure, which takes
      about 15 minutes, and causes a theoretical risk for perforations (1:1000) and an infection
      risk of 1:1.800.000, with lower risk for healthy subjects. The complication is serious and
      subjects will have to stay in the hospital. If there are serious complaints after the
      gastroscopy (e.g. vomiting with or without blood, or stomach ache) subjects are have to warn
      the project leader or their physician.

      Total time investment for the subjects will be approximately 14.5 hours, and 17.5 hours in
      the subgroup from which biopsies will be taken, excluding travel time.
allocation: Randomized
intervention model: Crossover Assignment
primary purpose: Prevention
masking: Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor)
measure: Flow mediated dilation
time frame: 4 weeks
description: FMD measurements after 4 weeks of intake. Fasting and postprandial measurements
measure: Intestinal apoA-I production
time frame: After 4 weeks of intake
description: Intestinal biopsies after 4 weeks of intake to see changes in apoA-I gene expression
measure: HDL cholesterol
time frame: 4 weeks
description: HDL serum cholesterol concentrations after 4 weeks of intake
measure: Vascular measurements
time frame: After 4 weeks
description: Vascular function measurements (PAT, PWV, PWA, retinal images) after 4 weeks of intake.
Fasting and postprandial measurements
measure: serum lipid metabolism
time frame: 4 weeks
description: serum lipid metabolism (total cholesterol, LDL-cholesterol, triglycerides) during fasting and in the postprandial state?
measure: glucose metabolism
time frame: 4 weeks
description: glucose and insulin concentrations during fasting and in the postprandial state?
measure: (apo)lipoprotein metabolism
time frame: 4 weeks
description: ApoB concentrations during fasting and in the postprandial state
arm group label: Placebo drink
arm group type: Placebo Comparator
description: A drink with the same components as the theobromine drink except for the theobromine
arm group label: Theobromine
arm group type: Experimental
description: 500mg theobromine in a drink
intervention type: Dietary Supplement
intervention name: Theobromine
description: Theobromine a compound for cocoa
arm group label: Theobromine
intervention type: Dietary Supplement
intervention name: Placebo drink
arm group label: Placebo drink
criteria: 
gender: All
minimum age: 45 Years
maximum age: 70 Years
healthy volunteers: Accepts Healthy Volunteers
last name: J Plat, Prof
role: Principal Investigator
affiliation: Maastricht University Hospital
facility: 
country: Netherlands
responsible party type: Sponsor
mesh term: Cardiovascular Diseases
mesh term: Theobromine