Impact of Beef on Metabolites and Inflammation
Condition: Healthy
MeSH terms: Inflammation
Study type: Interventional

Summary: 
      Western omnivore diets tend to use beef as a good source of protein. Studies have shown
      cattle who are fed different diets results in varying nutritional meat profiles in terms of
      amino acids, types and relative amounts of fat, and other nutrients. While overlap between
      organic and conventionally raised beef is expected, dietary impacts on human health are
      potentially meaningful as small changes in diet may have large downstream effects on human
      metabolism. To date, no studies have been conducted in humans that evaluate the health
      effects of organic grass-fed beef consumption to conventionally-fed beef consumption. To fill
      this gap in research, the investigators propose to identify metabolic differences in response
      to consumption of organic grass-fed compared to conventional-fed beef on a wide array of
      blood borne nutrients including amino acids, lipids, bile acids, and hundreds of other
      metabolites. This comprehensive analysis is expected to differentiate nutritional and
      metabolic impacts relevant to human health and provide a foundation for future research.

      The purpose of this study is to determine how a beef steak meal may affect bile acids in the
      gut, inflammation, and the metabolic health of healthy individuals. This research will
      investigate the following questions:

        1. How do specific foods affect bile acids in the blood?

        2. How do specific foods changes inflammation, metabolism, and other health measures? If
           researchers learn how food affects different health related markers in the blood and how
           that may influence health, then this information can be used to do more research to
           improve the health of people in a future study.
    

Detailed description: 
      The approach for this study is to enroll a population of healthy individuals (n=10; 5 men and
      5 women) in a double-blind, two-armed (Organic Grass-fed and Conventional-fed), randomized,
      crossover trial in which each individual participant serves as their own control to compare
      postprandial serum bile acid responses between steak meals, one which is organically
      grass-fed (OGF) and the other conventionally-fed (CONVF). Participants will consume a 6-oz
      steak meal grilled to 158°F, the recommended temperature by the USDA. The randomized
      crossover design with half of the participants starting in the OGF arm and the other half
      starting in the CONVF arm will eliminate potential order effects. Block randomization with 2
      blocks (total of 5 per block) will be created. Having each person serve as their own control
      in a crossover trial will eliminate the influence of inter-individual variation. The MSU
      Nutrition Research team has extensive experience with postprandial testing protocols and
      dietary manipulation and will utilize this experience to perform the proposed protocol with
      appropriate pre-test standardization, uniformity of meal preparation, precise timing of blood
      collection, and established procedures for processing and analyzing blood samples. Bile
      acids, including all primary and secondary forms, will be analyzed from samples collected
      before the meal and at 30-minute intervals for 4 hours after the meal. Established methods in
      the MSU Proteomics, Metabolomics, and Mass Spectrometry Facility will be used to compare
      total and individual bile acids between OGF and CONVF conditions.

      Procedures:

      During Visit 1, participants will undergo informed consent, be asked to complete a series of
      written questionnaires and anthropometric measurements. There will be a minimum 72 run-in
      period just prior to Visit 2, during which participants will be asked to follow certain
      dietary guidelines, such as no red meat. The second (Visit 2) and third (Visit 3) visits will
      take approximately 5 hours each and will each include a total of 9 blood draws through a
      sampling catheter, with each blood draw collecting about 2-3 teaspoons of blood. Blood will
      be drawn before a provided steak meal then every thirty minutes for 4 hours after meal
      consumption. There are a minimum of 72-hours between visits 1 and 2, and 7 days between
      visits 2 and 3. During this minimum 72- hour and 7-day period, participants will be asked to
      follow provided dietary and physical activity instructions. In addition, participants will be
      asked to complete a one month diet history questionnaire and a 24-hour dietary recall.

      Dietary intervention: A meat science specialist will select and prepare the steaks which will
      be given to participants. Participants will be asked to consume a 6-ounce beef steak meal,
      cooked to 158°F that is either OGF or CONVF, but will be coded and labeled to ensure
      blinding.

      Kinetics (measures over time) and untargeted and targeted metabolomics (an advance analytical
      technique to measure 1000's of molecules in a single sample) will be used as a tool to
      differentiate potential health impacts of the separate meal tests to investigate the
      influence of organic grass-fed beef versus conventionally-fed beef consumption on fasting and
      postprandial responses in healthy adults. The investigators expect to identify an array of
      metabolic differences between beef samples and will identify known impacts of those
      differences on health to provide a substantive comparison and provide a foundation for future
      research.

      Metabolomic and Lipidomic analysis: When all samples have been collected, the serum samples
      will be thawed, and a metabolite fraction will be extracted. After extraction, the samples
      will be explored using mass spectrometry analysis. An initial targeted analysis will focus on
      bile acids, lipids, and amino acids. Targeted analytes will be quantified using standard
      curves and the resulting concentrations will be compared using several statistical analysis
      tools. Metabolite samples will be analyzed using a Waters Synapt G2Si QTOF (Waters
      Corporation). This instrument has ion mobility capability and can differentiate closely
      related compounds, such as bile acids, in complex mixtures based on their cross-sectional
      area and mass. The high sensitivity and variety of scan modes make the Synapt QTOF the
      optimal choice for untargeted analysis of complex biological samples.

      Dietary analysis: This study will use the most recent version (2018) of the web-based Diet
      History Questionnaire (DHQ III), a food frequency questionnaire designed for adults 19 and
      older, developed by staff at the Risk Factor Monitoring and Methods Branch (RFMMB) of the NIH
      National Cancer Institute. The outputs of the DHQ III include carbohydrate constituents,
      carotenoids and tocopherols, dietary constituents from supplements, fats, fatty acids and
      cholesterol, macronutrients and energy, minerals, protein constituents, and vitamins are
      dietary constituents and food groups available in the DHQ III output files. In addition,
      acute dietary consumption will be determined using the free, web-based Automated
      Self-Administered 24-hour (ASA24®) questionnaire.
    

Intervention name(s): Conventional-fed beef; Organic grass-fed beef
Intervention type(s): Behavioral; Behavioral
Intervention description(s): Participants in the conventional-fed treatment of the study will consume a conventional-fed steak meal for breakfast one morning.; Participants in the organic grass-fed treatment of the study will consume a organic grass-fed steak meal for breakfast one morning.
Allocation: Randomized
Intervention model: Crossover Assignment
Intervention model description: The design is 2 x 2 crossover design, where the healthy adults will be randomized into Organic grass-fed and conventional-fed conditions for the first round of data collection and then crossover to the other condition for the second round of data collection.
Masking: Double (Participant, Investigator)
Primary purpose: Prevention
Keywords: Organic grass-fed; Conventional-fed