<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/NCT03752125</url>
  </required_header>
  <id_info>
    <org_study_id>18-0257</org_study_id>
    <nct_id>NCT03752125</nct_id>
  </id_info>
  <brief_title>Influence of Flavonoids and Caffeine Supplementation on Resting Metabolism</brief_title>
  <official_title>A Randomized, Cross-Over Clinical Trial to Evaluate the Acute Effects of Mixed Flavonoid and Caffeine Ingestion on Energy Expenditure and Fat Oxidation in Healthy Adult Women</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>Appalachian State University</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>University of North Carolina, Chapel Hill</agency>
      <agency_class>Other</agency_class>
    </collaborator>
  </sponsors>
  <source>Appalachian State University</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>
      The purpose of this study is to compare the effect of ingesting caffeine and mixed flavonoids&#xD;
      (4 capsules, split between breakfast and lunch) on energy expenditure and fat oxidation in a&#xD;
      metabolic chamber with 20 women (non-obese, healthy, ages 20-47 years, pre-menopausal). We&#xD;
      hypothesize that based on the existing literature, ingestion of a double dose of the&#xD;
      caffeine-mixed flavonoid supplement compared to placebo will increase fat oxidation and&#xD;
      increase 24 h energy expenditure by about 75 kilocalories.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      This study will utilize a randomized, double-blind, cross-over design, comparing acute&#xD;
      ingestion of the mixed flavonoid supplement (MF) compared to placebo control (PC) in healthy,&#xD;
      non-obese, pre-menopausal women. The study will consist of two 24-hour study periods spent in&#xD;
      the recently renovated indirect room calorimeter (i.e., metabolic chamber) at the University&#xD;
      of North Carolina Chapel Hill Nutrition Research Institute (UNC NRI). The 24-hour study&#xD;
      periods in the metabolic chamber will be 4 weeks apart. Primary outcome measures will be&#xD;
      24-hour energy expenditure (EE), resting metabolic rate (RMR), sleeping metabolic rate (SMR),&#xD;
      substrate utilization from the respiratory quotient (RQ), and physical activity energy&#xD;
      expenditure (AEE).&#xD;
&#xD;
      Following recruitment and eligibility assessment, study participants will be randomly&#xD;
      assigned to complete either the mixed flavonoid (MF) or the placebo control (PC) study day.&#xD;
      After a 4-week washout, study participants will cross over and complete the alternative&#xD;
      treatment. A method of randomly permuted blocks will be generated using web-based&#xD;
      randomization software (www.randomization.com) resulting in 10 study participants receiving&#xD;
      MF during the first week, and 10 study participants receiving the PC during the first week.&#xD;
      Study participants will consume 2 MF or PC capsules 30 min before breakfast, and 2 MF or PC&#xD;
      capsules 30 min before lunch. During both study days, study participants will be fed in&#xD;
      energy balance (i.e., energy intake will be matched to energy expenditure).&#xD;
&#xD;
      2. Experimental Study Participants: Using data generated previously in the UNC NRI metabolic&#xD;
      chamber, a sample size calculation with 80% power revealed that 15-20 study participants&#xD;
      would be needed to detect a 50 kcal difference in 24-h EE. To account for attrition, 25 study&#xD;
      participants will be recruited via mass advertisement throughout the local area.&#xD;
      Pre-menopausal women will be chosen because this is a target population of the sponsor.&#xD;
&#xD;
      3. Pre-Study Baseline Testing Eligibility will be determined in the outpatient clinical suite&#xD;
      at the UNC NRI. Body composition (fat mass and fat free mass (FFM)) will be determined via&#xD;
      dual energy x-ray absorptiometry (DXA) (GE Lunar iDXA; Milwaukee, WI). Body mass index (BMI,&#xD;
      kg/m2) will be calculated for measured height and weight. Resting metabolic rate (RMR) will&#xD;
      be estimated using a FFM-based equation [418 + (20.3 FFM)] (J Appl Physiol&#xD;
      1993;75:2514-2520). This estimated RMR will be used to calculate total dietary energy intake&#xD;
      while in the metabolic chamber: RMR x physical activity level (PAL) of 1.3, and then adjusted&#xD;
      using measured data (details provided below). A small blood sample will be obtained at this&#xD;
      time and T3, T4, and TSH levels assessed through the clinical lab at the Lab Corp&#xD;
      (Burlington, NC). A urine pregnancy test will be performed.&#xD;
&#xD;
      4. Indirect calorimetry: The metabolic chamber at the Nutrition Research Institute in&#xD;
      Kannapolis, NC is an open-circuit, whole room indirect calorimeter. The CO2 and O2 analysers&#xD;
      are differential, with full scale readings set for 0%-1%. O2 consumption, CO2 production, EE&#xD;
      and RQ are recorded each minute. EE is calculated using an abbreviated Weir's formula (VO2 X&#xD;
      3.941) + (VCO2 X 1.106), where VO2 is the volume of oxygen consumed in L/minute and VCO2 is&#xD;
      the volume of carbon dioxide released in L/minute. RQ is calculated as VCO2/VO2. Area under&#xD;
      the curve (AUC) will be calculated using RQ data for the four hours following breakfast,&#xD;
      lunch and dinner as well as sleeping hours between midnight and 6:00 am. Spontaneous physical&#xD;
      activity will be measured each minute using a total room sensor. To calculate resting&#xD;
      metabolic rate (RMR), EE will be plotted against the activity motion sensor output (each&#xD;
      averaged over 30 minutes), and the y-intercept of the linear regression taken as EE in the&#xD;
      inactive state. RMR will be multiplied by 1440 minutes to extrapolate to 24 hours.&#xD;
      Twenty-four hour sleeping metabolic rate (SMR) will be determined as the lowest mean EE&#xD;
      (kJ/minute) measured over 3 consecutive hours between midnight and 6:30 am and multiplied by&#xD;
      1440 minutes. Diet induced thermogenesis (DIT) will be calculated by subtraction of SMR from&#xD;
      RMR. Activity induced EE (AEE) will be calculated as the difference between 24 hour EE and&#xD;
      RMR.&#xD;
&#xD;
      5. Metabolic chamber study day protocol. Subjects will arrive at the Nutrition Research&#xD;
      Institute Building (500 Laureate Way) at 7:00 am. A urine pregnancy test will be performed.&#xD;
      At 7:30 am, study participants will report to the metabolic chamber following an overnight&#xD;
      fast (no food or beverages containing calories, alcohol, or caffeine from 11 pm). Study&#xD;
      participants will be instructed on expectations of their stay and weighed in scrubs without&#xD;
      shoes. At 8:00 am study participants will be sealed in the chamber. Except for a 2-minute&#xD;
      interval each hour during which study participants will be requested to stand and stretch,&#xD;
      study participants will remain seated or reclined, but awake throughout the day. Study&#xD;
      participants will be asked to perform necessary daily activities during these 2-minute&#xD;
      intervals. Breakfast (9:00 am), lunch (1:30 pm), and dinner (7:00 pm), will be served through&#xD;
      an air-lock passage. Meals will be completed within 30 minutes of serving. Two MF capsules or&#xD;
      placebo will be consumed 30 min before breakfast, and then again 30 minutes before lunch. At&#xD;
      10:30 pm, study participants will be asked to lie down for sleep. Study participants will be&#xD;
      awakened at 6:30 am and allowed to move about the chamber to gather their belongings. At 7:15&#xD;
      am study participants will exit the chamber and be weighed.&#xD;
&#xD;
      6. Design of Metabolic Diets: Eucaloric diets will be designed to provide approximately 35%&#xD;
      fat, 49% carbohydrates and 16% protein, reflecting current recommendations for this&#xD;
      population group. Menus will be designed using a nutrient calculation and food management&#xD;
      software, and consist of bagel, peanut butter, apple juice, whole-wheat bread, turkey,&#xD;
      cheese, mayonnaise, buttery spread (10% kcal as fat), potato chips, lasagna, carrots,&#xD;
      broccoli, rolls, and muffin. No beverages or foods containing caffeine will be served. The&#xD;
      same foods will be served at both chamber visits. A baseline menu for each subject will be&#xD;
      prepared based on calculated RMR X 1.3, reflecting the sedentary nature of the study day. To&#xD;
      ensure energy balance conditions, the baseline menu of the first visit will be modified&#xD;
      according to measured EE data at 3 hours (includes breakfast) and 7 hours (includes breakfast&#xD;
      and lunch). Subsequent meals will be adjusted accordingly with 100 kcal peach muffins&#xD;
      containing the same proportion of fat, carbohydrate and protein as the meals. Study&#xD;
      participants will be fed an identical amount of the same meal at their second visit.&#xD;
&#xD;
      7. Statistical Analysis: Data will be analyzed using SAS (Cary, NC). To guard against any&#xD;
      carryover effect from visit 1 to visit 2, a repeated measures regression with an unstructured&#xD;
      correlation matrix within subject will be run with treatment (MF and PC) and visit (1 and 2)&#xD;
      in the model along with an interaction term. If the interaction term is non-significant, then&#xD;
      it will be removed and the model re-run. Two energy expenditure curves, one for the MF day,&#xD;
      and one for the PC day, will be generated for each subject with the x axis representing time&#xD;
      (min), and the y axis representing energy expenditure (kcal). The area under the energy&#xD;
      expenditure curve for defined blocks of time will be calculated by using the trapezoid rule&#xD;
      in the EXPAND procedure in SAS (SAS Institute, Inc., Cary, NC). A paired t-test on the&#xD;
      log-transformed area will be performed to compare the energy expenditure of each period in&#xD;
      the MF day with the corresponding period in the PC day. The Shapiro-Wilk test in the&#xD;
      UNIVARIATE procedure in SAS will be used for normality check. The Benjamini-Hochberg method&#xD;
      for false discovery rate correction in the MULTTEST procedure in SAS will be used for&#xD;
      multiple testing corrections.&#xD;
&#xD;
      SUPPLEMENT:&#xD;
&#xD;
      The mixed flavonoid supplement and placebo capsules will look identical and be supplied by&#xD;
      Reoxcyn Discoveries Group LLC (Salt Lake City, UT). Active ingredients in the flavonoid&#xD;
      capsules include vitamin C, wild bilberry fruit extract, green tea leaf extract, quercetin,&#xD;
      caffeine, and omega 3 fatty acids (see Appendix B). Capsule fill ingredients include Nu-Flow&#xD;
      70R (from powdered rice hulls), tapioca from cassava root, natural bamboo silica, and&#xD;
      marshmallow root. Placebo capsules will contain only the fill ingredients (without the active&#xD;
      ingredients). Two MF and PC capsules will be consumed 30 minutes before breakfast, and then&#xD;
      again before lunch. The MF capsules will provide 658 mg flavonoids and 214 mg caffeine.&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Completed</overall_status>
  <start_date type="Actual">April 22, 2018</start_date>
  <completion_date type="Actual">April 19, 2019</completion_date>
  <primary_completion_date type="Actual">December 15, 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>Randomized, crossover</intervention_model_description>
    <primary_purpose>Treatment</primary_purpose>
    <masking>Double (Participant, Investigator)</masking>
    <masking_description>Flavonoid and placebo capsules</masking_description>
  </study_design_info>
  <primary_outcome>
    <measure>24-hour energy expenditure (kilocalories/minute) from continuous respiratory exchange measurements (oxygen consumption, carbon dioxide production) in a metabolic chamber</measure>
    <time_frame>24 hours</time_frame>
    <description>Energy expenditure (kilocalories) over 24 h in metabolic chamber with oxygen consumption and carbon dioxide production analyzers whereby energy expenditure is calculated from respiratory exchange data (oxygen consumption, carbon dioxide production) using standard equations</description>
  </primary_outcome>
  <secondary_outcome>
    <measure>24-hour fat oxidation (fat grams oxidized per minute) from continuous respiratory exchange measurements (oxygen consumption, carbon dioxide production) in a metabolic chamber</measure>
    <time_frame>24 hours</time_frame>
    <description>Fat oxidation (fat grams) over 24 h in metabolic chamber with oxygen consumption and carboh dioxide production analyzers whereby fat oxidation (grams/minute) is calculated from respiratory exchange data (oxygen consumption, carbon dioxide production) using standard equations</description>
  </secondary_outcome>
  <number_of_arms>2</number_of_arms>
  <enrollment type="Actual">25</enrollment>
  <condition>Diet Modification</condition>
  <arm_group>
    <arm_group_label>Flavonoid, caffeine</arm_group_label>
    <arm_group_type>Experimental</arm_group_type>
    <description>Flavonoid, caffeine capsules</description>
  </arm_group>
  <arm_group>
    <arm_group_label>Placebo</arm_group_label>
    <arm_group_type>Placebo Comparator</arm_group_type>
    <description>Placebo capsules</description>
  </arm_group>
  <intervention>
    <intervention_type>Dietary Supplement</intervention_type>
    <intervention_name>Flavonoid, caffeine</intervention_name>
    <description>2 flavonoid/caffeine capsules before breakfast, 2 capsules before lunch</description>
    <arm_group_label>Flavonoid, caffeine</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Dietary Supplement</intervention_type>
    <intervention_name>Placebo</intervention_name>
    <description>2 placebo capsules before breakfast, 2 capsules before lunch</description>
    <arm_group_label>Placebo</arm_group_label>
  </intervention>
  <eligibility>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  Women, ages 20 to 47 years&#xD;
&#xD;
          -  Body mass index (BMI) between 18.5 and 33 kg/m2 (normal weight, overweight, mild&#xD;
             obesity)&#xD;
&#xD;
          -  Weight during the past 6 months has been somewhat stable (+/- 12 lbs)&#xD;
&#xD;
          -  Low-to-moderate caffeine intake, defined as consuming on average: less than 3 servings&#xD;
             per day of coffee (each about 8 fl. oz.) and/or energy drinks with caffeine; less than&#xD;
             3 soft drinks with caffeine and tea per day&#xD;
&#xD;
          -  Premenopausal (self-report)&#xD;
&#xD;
          -  Willingness to maintain habitual diet and physical activity patterns, and not lose&#xD;
             weight throughout the study period&#xD;
&#xD;
          -  Understands and agrees to all study procedures and signs the form providing informed&#xD;
             consent to participate in the study&#xD;
&#xD;
          -  Willingness to report and maintain normal schedule of hormonal therapy including oral&#xD;
             contraceptive pills, hormonal IUD, Nuva Ring, or DepoProvera injections.&#xD;
&#xD;
          -  Normal thyroid function as determined from a blood sample collected during pre-study&#xD;
             baseline testing.&#xD;
&#xD;
          -  Not pregnant as determined by a urine pregnancy test during pre-study baseline testing&#xD;
             and prior to the second chamber session, and not planning to become pregnant during&#xD;
             the study.&#xD;
&#xD;
          -  Willingness to avoid vigorous exercise the day before each metabolic chamber session&#xD;
             (e.g., intense running, cycling, swimming for 30 minutes and longer).&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  Taking medications for or suffering from a medical condition that may impact results&#xD;
             related to metabolism (e.g. thyroid disorders, diabetes, mental disorders such as&#xD;
             anxiety or depression, heart disease, arthritis, cancer).&#xD;
&#xD;
          -  Heavy exerciser (defined as more than 300 minutes per week of moderate to vigorous&#xD;
             exercise within the last two weeks)&#xD;
&#xD;
          -  Thyroid hormone levels outside of the normal laboratory reference range&#xD;
&#xD;
          -  Pregnant or lactating, or planning to become pregnant during the study&#xD;
&#xD;
          -  Recent use of antibiotics (within last two weeks)&#xD;
&#xD;
          -  History of smoking during the six months prior to study&#xD;
&#xD;
          -  Recent signs or symptoms of infection, including cold or flu-like symptoms (within&#xD;
             last 2 weeks)&#xD;
&#xD;
          -  Conditions or diseases the investigator believes would interfere with ability to&#xD;
             provide consent, comply with the study protocol, which might confound the&#xD;
             interpretation of the study results or put the person at undue risk. These include the&#xD;
             disease list above and other health problems that would make it difficult to comply&#xD;
             with two 24-hour periods of confinement in the metabolic chamber such as panic&#xD;
             disorder, cardiac arrhythmias, musculoskeletal disorders such as low back pain and&#xD;
             osteoarthritis, and gastrointestinal disorders including reactions to supplements with&#xD;
             caffeine and vitamin C, and food allergies.&#xD;
&#xD;
          -  Inability or unwillingness to consume the foods that will be included in the provided&#xD;
             meals when staying in the chamber&#xD;
&#xD;
          -  History of drug or alcohol abuse (&gt;5 standard drinks per day)&#xD;
&#xD;
          -  Taking herbal supplements or medications that may influence metabolism within 14 days&#xD;
             prior to the study, including supplements containing bitter orange, guarana,&#xD;
             fucoxanthin, and green coffee bean, and medications such as Meridia, Xenical,&#xD;
             LipoRidAM, Advantra-Z, NuPhedrine, and Slim-10.&#xD;
&#xD;
          -  Current consumption (or unwilling to stop intake 2 weeks prior) of flavonoid&#xD;
             supplements, or a heavy consumer of green tea (&gt;3 cups per day)&#xD;
&#xD;
          -  Near-daily use of tablets containing caffeine (e.g., NoDoz) (or unwillingness to stop&#xD;
             2 weeks prior to the study).&#xD;
&#xD;
          -  High intake of caffeine from coffee, tea, energy drinks with caffeine, and soft drinks&#xD;
             containing caffeine (or unwillingness to reduce intake for 2 weeks prior to the study&#xD;
             and during the 4-week study).&#xD;
&#xD;
          -  Prone to panic attacks when confined in a small room, especially when consuming&#xD;
             caffeine.&#xD;
      </textblock>
    </criteria>
    <gender>Female</gender>
    <gender_based>Yes</gender_based>
    <minimum_age>20 Years</minimum_age>
    <maximum_age>47 Years</maximum_age>
    <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>David C Nieman, DrPH</last_name>
    <role>Principal Investigator</role>
    <affiliation>Appalachian State Univ</affiliation>
  </overall_official>
  <location>
    <facility>
      <name>Appalachian State University Human Performance Lab, North Carolina Research Campus</name>
      <address>
        <city>Kannapolis</city>
        <state>North Carolina</state>
        <zip>28081</zip>
        <country>United States</country>
      </address>
    </facility>
  </location>
  <location_countries>
    <country>United States</country>
  </location_countries>
  <verification_date>July 2019</verification_date>
  <study_first_submitted>May 3, 2018</study_first_submitted>
  <study_first_submitted_qc>November 21, 2018</study_first_submitted_qc>
  <study_first_posted type="Actual">November 23, 2018</study_first_posted>
  <last_update_submitted>July 3, 2019</last_update_submitted>
  <last_update_submitted_qc>July 3, 2019</last_update_submitted_qc>
  <last_update_posted type="Actual">July 5, 2019</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor</responsible_party_type>
  </responsible_party>
  <keyword>energy expenditure</keyword>
  <keyword>caffeine</keyword>
  <keyword>flavonoids</keyword>
  <keyword>fat metabolism</keyword>
  <intervention_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Caffeine</mesh_term>
  </intervention_browse>
  <patient_data>
    <sharing_ipd>No</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/NCT03752125
org study id: 18-0257
nct id: NCT03752125
lead sponsor: 
collaborator: 
has dmc: No
is fda regulated drug: No
is fda regulated device: No
The purpose of this study is to compare the effect of ingesting caffeine and mixed flavonoids
      (4 capsules, split between breakfast and lunch) on energy expenditure and fat oxidation in a
      metabolic chamber with 20 women (non-obese, healthy, ages 20-47 years, pre-menopausal). We
      hypothesize that based on the existing literature, ingestion of a double dose of the
      caffeine-mixed flavonoid supplement compared to placebo will increase fat oxidation and
      increase 24 h energy expenditure by about 75 kilocalories.
This study will utilize a randomized, double-blind, cross-over design, comparing acute
      ingestion of the mixed flavonoid supplement (MF) compared to placebo control (PC) in healthy,
      non-obese, pre-menopausal women. The study will consist of two 24-hour study periods spent in
      the recently renovated indirect room calorimeter (i.e., metabolic chamber) at the University
      of North Carolina Chapel Hill Nutrition Research Institute (UNC NRI). The 24-hour study
      periods in the metabolic chamber will be 4 weeks apart. Primary outcome measures will be
      24-hour energy expenditure (EE), resting metabolic rate (RMR), sleeping metabolic rate (SMR),
      substrate utilization from the respiratory quotient (RQ), and physical activity energy
      expenditure (AEE).

      Following recruitment and eligibility assessment, study participants will be randomly
      assigned to complete either the mixed flavonoid (MF) or the placebo control (PC) study day.
      After a 4-week washout, study participants will cross over and complete the alternative
      treatment. A method of randomly permuted blocks will be generated using web-based
      randomization software (www.randomization.com) resulting in 10 study participants receiving
      MF during the first week, and 10 study participants receiving the PC during the first week.
      Study participants will consume 2 MF or PC capsules 30 min before breakfast, and 2 MF or PC
      capsules 30 min before lunch. During both study days, study participants will be fed in
      energy balance (i.e., energy intake will be matched to energy expenditure).

      2. Experimental Study Participants: Using data generated previously in the UNC NRI metabolic
      chamber, a sample size calculation with 80% power revealed that 15-20 study participants
      would be needed to detect a 50 kcal difference in 24-h EE. To account for attrition, 25 study
      participants will be recruited via mass advertisement throughout the local area.
      Pre-menopausal women will be chosen because this is a target population of the sponsor.

      3. Pre-Study Baseline Testing Eligibility will be determined in the outpatient clinical suite
      at the UNC NRI. Body composition (fat mass and fat free mass (FFM)) will be determined via
      dual energy x-ray absorptiometry (DXA) (GE Lunar iDXA; Milwaukee, WI). Body mass index (BMI,
      kg/m2) will be calculated for measured height and weight. Resting metabolic rate (RMR) will
      be estimated using a FFM-based equation [418 + (20.3 FFM)] (J Appl Physiol
      1993;75:2514-2520). This estimated RMR will be used to calculate total dietary energy intake
      while in the metabolic chamber: RMR x physical activity level (PAL) of 1.3, and then adjusted
      using measured data (details provided below). A small blood sample will be obtained at this
      time and T3, T4, and TSH levels assessed through the clinical lab at the Lab Corp
      (Burlington, NC). A urine pregnancy test will be performed.

      4. Indirect calorimetry: The metabolic chamber at the Nutrition Research Institute in
      Kannapolis, NC is an open-circuit, whole room indirect calorimeter. The CO2 and O2 analysers
      are differential, with full scale readings set for 0%-1%. O2 consumption, CO2 production, EE
      and RQ are recorded each minute. EE is calculated using an abbreviated Weir's formula (VO2 X
      3.941) + (VCO2 X 1.106), where VO2 is the volume of oxygen consumed in L/minute and VCO2 is
      the volume of carbon dioxide released in L/minute. RQ is calculated as VCO2/VO2. Area under
      the curve (AUC) will be calculated using RQ data for the four hours following breakfast,
      lunch and dinner as well as sleeping hours between midnight and 6:00 am. Spontaneous physical
      activity will be measured each minute using a total room sensor. To calculate resting
      metabolic rate (RMR), EE will be plotted against the activity motion sensor output (each
      averaged over 30 minutes), and the y-intercept of the linear regression taken as EE in the
      inactive state. RMR will be multiplied by 1440 minutes to extrapolate to 24 hours.
      Twenty-four hour sleeping metabolic rate (SMR) will be determined as the lowest mean EE
      (kJ/minute) measured over 3 consecutive hours between midnight and 6:30 am and multiplied by
      1440 minutes. Diet induced thermogenesis (DIT) will be calculated by subtraction of SMR from
      RMR. Activity induced EE (AEE) will be calculated as the difference between 24 hour EE and
      RMR.

      5. Metabolic chamber study day protocol. Subjects will arrive at the Nutrition Research
      Institute Building (500 Laureate Way) at 7:00 am. A urine pregnancy test will be performed.
      At 7:30 am, study participants will report to the metabolic chamber following an overnight
      fast (no food or beverages containing calories, alcohol, or caffeine from 11 pm). Study
      participants will be instructed on expectations of their stay and weighed in scrubs without
      shoes. At 8:00 am study participants will be sealed in the chamber. Except for a 2-minute
      interval each hour during which study participants will be requested to stand and stretch,
      study participants will remain seated or reclined, but awake throughout the day. Study
      participants will be asked to perform necessary daily activities during these 2-minute
      intervals. Breakfast (9:00 am), lunch (1:30 pm), and dinner (7:00 pm), will be served through
      an air-lock passage. Meals will be completed within 30 minutes of serving. Two MF capsules or
      placebo will be consumed 30 min before breakfast, and then again 30 minutes before lunch. At
      10:30 pm, study participants will be asked to lie down for sleep. Study participants will be
      awakened at 6:30 am and allowed to move about the chamber to gather their belongings. At 7:15
      am study participants will exit the chamber and be weighed.

      6. Design of Metabolic Diets: Eucaloric diets will be designed to provide approximately 35%
      fat, 49% carbohydrates and 16% protein, reflecting current recommendations for this
      population group. Menus will be designed using a nutrient calculation and food management
      software, and consist of bagel, peanut butter, apple juice, whole-wheat bread, turkey,
      cheese, mayonnaise, buttery spread (10% kcal as fat), potato chips, lasagna, carrots,
      broccoli, rolls, and muffin. No beverages or foods containing caffeine will be served. The
      same foods will be served at both chamber visits. A baseline menu for each subject will be
      prepared based on calculated RMR X 1.3, reflecting the sedentary nature of the study day. To
      ensure energy balance conditions, the baseline menu of the first visit will be modified
      according to measured EE data at 3 hours (includes breakfast) and 7 hours (includes breakfast
      and lunch). Subsequent meals will be adjusted accordingly with 100 kcal peach muffins
      containing the same proportion of fat, carbohydrate and protein as the meals. Study
      participants will be fed an identical amount of the same meal at their second visit.

      7. Statistical Analysis: Data will be analyzed using SAS (Cary, NC). To guard against any
      carryover effect from visit 1 to visit 2, a repeated measures regression with an unstructured
      correlation matrix within subject will be run with treatment (MF and PC) and visit (1 and 2)
      in the model along with an interaction term. If the interaction term is non-significant, then
      it will be removed and the model re-run. Two energy expenditure curves, one for the MF day,
      and one for the PC day, will be generated for each subject with the x axis representing time
      (min), and the y axis representing energy expenditure (kcal). The area under the energy
      expenditure curve for defined blocks of time will be calculated by using the trapezoid rule
      in the EXPAND procedure in SAS (SAS Institute, Inc., Cary, NC). A paired t-test on the
      log-transformed area will be performed to compare the energy expenditure of each period in
      the MF day with the corresponding period in the PC day. The Shapiro-Wilk test in the
      UNIVARIATE procedure in SAS will be used for normality check. The Benjamini-Hochberg method
      for false discovery rate correction in the MULTTEST procedure in SAS will be used for
      multiple testing corrections.

      SUPPLEMENT:

      The mixed flavonoid supplement and placebo capsules will look identical and be supplied by
      Reoxcyn Discoveries Group LLC (Salt Lake City, UT). Active ingredients in the flavonoid
      capsules include vitamin C, wild bilberry fruit extract, green tea leaf extract, quercetin,
      caffeine, and omega 3 fatty acids (see Appendix B). Capsule fill ingredients include Nu-Flow
      70R (from powdered rice hulls), tapioca from cassava root, natural bamboo silica, and
      marshmallow root. Placebo capsules will contain only the fill ingredients (without the active
      ingredients). Two MF and PC capsules will be consumed 30 minutes before breakfast, and then
      again before lunch. The MF capsules will provide 658 mg flavonoids and 214 mg caffeine.
allocation: Randomized
intervention model: Crossover Assignment
intervention model description: Randomized, crossover
primary purpose: Treatment
masking: Double (Participant, Investigator)
masking description: Flavonoid and placebo capsules
measure: 24-hour energy expenditure (kilocalories/minute) from continuous respiratory exchange measurements (oxygen consumption, carbon dioxide production) in a metabolic chamber
time frame: 24 hours
description: Energy expenditure (kilocalories) over 24 h in metabolic chamber with oxygen consumption and carbon dioxide production analyzers whereby energy expenditure is calculated from respiratory exchange data (oxygen consumption, carbon dioxide production) using standard equations
measure: 24-hour fat oxidation (fat grams oxidized per minute) from continuous respiratory exchange measurements (oxygen consumption, carbon dioxide production) in a metabolic chamber
time frame: 24 hours
description: Fat oxidation (fat grams) over 24 h in metabolic chamber with oxygen consumption and carboh dioxide production analyzers whereby fat oxidation (grams/minute) is calculated from respiratory exchange data (oxygen consumption, carbon dioxide production) using standard equations
arm group label: Flavonoid, caffeine
arm group type: Experimental
description: Flavonoid, caffeine capsules
arm group label: Placebo
arm group type: Placebo Comparator
description: Placebo capsules
intervention type: Dietary Supplement
intervention name: Flavonoid, caffeine
description: 2 flavonoid/caffeine capsules before breakfast, 2 capsules before lunch
arm group label: Flavonoid, caffeine
intervention type: Dietary Supplement
intervention name: Placebo
description: 2 placebo capsules before breakfast, 2 capsules before lunch
arm group label: Placebo
criteria: 
gender: Female
gender based: Yes
minimum age: 20 Years
maximum age: 47 Years
healthy volunteers: Accepts Healthy Volunteers
last name: David C Nieman, DrPH
role: Principal Investigator
affiliation: Appalachian State Univ
facility: 
country: United States
responsible party type: Sponsor
mesh term: Caffeine
sharing ipd: No