https://cris.nifa.usda.gov/cgi-bin/starfinder/130833/crisassist.

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ACCESSION NO: 1010729 SUBFILE: CRIS
PROJ NO: IND030455 AGENCY: NIFA IND
PROJ TYPE: HATCH PROJ STATUS: TERMINATED
START: 20 OCT 2016 TERM: 02 OCT 2019 FY: 2018

INVESTIGATOR: Mattes, RI, .

PERFORMING INSTITUTION:
PURDUE UNIVERSITY
WEST LAFAYETTE, INDIANA 47907

LOW CALORIE SWEETENERS: HEDONICS, INTAKE AND METABOLISM

NON-TECHNICAL SUMMARY: The objectives of this application are to

elucidate selected mechanisms and functions for an array of LCS including
the most widely used varieties (sucralose, aspartame, saccharin, stevia) as
they relate to ingestive behavior and glycemia. The long-term goal is to clarify
the role of LCS in weight management regimens and blood sugar
management. Our rationale is that HIS are widely consumed and often
recommended to reduce energy intake and post-prandial glycemia yet there
are allegations that they may be ineffective for these purposes or actually
promote energy intake and disorder glycemia. If the adverse outcomes are
true, dietary recommendations will require modification.

OBJECTIVES: Contrast the effects of the four most widely used LCS

(aspartame, saccharin, sucralose, stevia) on body weight and
composition.Determine whether acute and chronic use of aspartame disrupts
glucose tolerance, appetite and gut hormone secretion in healthy adults.

APPROACH: Two trials are proposed, one will explore effects of LCS on body
weight and the other will focus on glycemia1. Evaluate the effects of the four
most widely-used HIS on appetite, energy intake, body weight and body
composition.Participants: 5 groups of 25, 18-60 yrs, with BMI between 25 and
40 kg/M2. Recruited by public advertisement.Baseline: Participants will report
to the lab after an overnight fast. A 2-hr OGTT will be given and the samples
will be analyzed for lipid profile, glucose and insulin. They will also receive
instruction about study procedures and requirements. They will learn to record
food and keep a record for 3 days and record appetitive sensations. These
ratings will be obtained over one 24-hr period during one day that they record
food intake. They will also be provided a RT6 wrist accelerometer to monitor
their daily physical activity for the same three days diet is recorded. Finally,
they will be provided urine containers with boric acid added as a preservative
for a 24-hr collection to determine their baseline PABA and urinary nitrogen
excretion. PABA will be added to the beverages and used to measure
compliance with the prescribed beverage ingestion and urinary nitrogen will
be used to validate the diet records as described in detail below. Urine will be
collected the final day of food intake recording and <48 hrs prior to the
participant's scheduled first intervention visit. At the end of the baseline week,
participants will return to the laboratory. They will return the accelerometer
and their urine sample, have weight measured and be randomly assigned to a
group that receives one of the four HIS or sucrose. They will then be
instructed to initiate consumption of their prescribed beverage daily. No
dietary advice will be provided other than that they are expected to consume
the provided beverage daily. They will be told that the purpose of the study is
to determine the effects of beverage consumption on well-being and selected
health outcomes. Measurements will be taken of blood pressure, grip strength
and fine motor control to minimize knowledge of weight as a primary outcome
on dietary behavior.Intervention: Participants will only be in one arm of the
study and consume only one HIS or sucrose. Participants will report to the
laboratory on a weekly basis for measurement of body weight; to receive
beverage packets; to reinforce the importance of complying with the study
procedures and to address any issues that may arise. Body composition and
biochemical indices (e.g., lipid profiles, glucose, insulin) will be assessed at
baseline and at the end of the trial. At weeks B, 4, 8 and 12, participants will
be asked to record food intake for 3 days, wear an accelerometer for the
same 3 days, document appetitive sensations hourly for all waking hours for
one of the days a diet record is kept and collect a 24-hr urine sample one day
a diet record is kept. By measuring the latter four outcomes concurrently, it will
be possible to ascertain the relationship between sweetener use, diet, energy
expenditure and appetite while confirming ingestion of the intervention
beverage and validating the diet records. Every two weeks, participants will
complete distractor tasks in the laboratory.Beverage: The sucrose and LCS
will be presented in beverages. The amount will be set at a portion that
provides 1.8g of sucrose/kg BW presented as a 9% sucrose
solution.Anthropometry: Height will be measured at baseline with a Holtain
stadiometer. Body weight will be measured weekly with participants in a
standard gown using a single calibrated clinical scale. Body composition will
be measured at baseline and week 12 by DEXA. Energy metabolism and
physical activity: Free-living physical activity levels and energy expenditure
will be predicted using RT6 triaxial accelerometers (StayHealthy, Monrovia,
CA). Free-living physical activity levels will be measured at baseline and
weeks 4, 8 and 12. Measurements will be taken over three consecutive days
that include two weekdays and one weekend day. Habitual physical activity
will also be assessed using a validated questionnaire.Dietary intake: Food and
energy intake will be assessed at baseline and weeks 4, 8 and 12 using the
web-based "Automated Self-Administered 24-hr Dietary Recall" (ASA24)
system. Participants will be asked to record dietary intake for three non-
consecutive days that include two non-consecutive weekdays and one
weekend day for better representation of habitual intake. It is well understood
that dietary data are subject to inaccurate reporting so they will be verified
against urinary nitrogen excretion measured in the collected urine
samples.Appetite: Ratings of hunger, fullness, desire to eat, desire to eat
something sweet, desire to eat something salty, prospective consumption and
thirst will be recorded at baseline and weeks 4, 8 and 12 (on the hour, every
hour while awake for one day) using validated visual analog scales
programmed into the participants phone using Qualtrics. Pattern, peak, nadir
and AUC values will be computedBiochemistry: A standard 2-hr (75 gram
carbohydrate load) OGTT will be administered at baseline and week 12. Four
milliliter blood sample will be obtained in a red top vacutainer at baseline and
minutes, 30, 60, 90 and 120. A single fasting blood draw will also be collected
at week 6 for a lipid panel and fasting insulin and glucose. Samples will be
centrifuged and the serum aliquotted and frozen at -80oC for batch analysis.
Triacylglycerol, high density and low density cholesterol and glucose
concentrations will be determined on a. Randox Daytona Clinical Analyzer
(CDMD Analyte Laboratory). The sensitivity of the assay is 0.12mmol/L and
the CV is ~1.0%. .Insulin will be measured using the Elecsys® 2010
Immunoassay System (Roche Diagnostic Systems, Indianapolis, IN). It has a
sensitivity of 0.20uU/ml and a CV of ~ 2.6%.Compliance: PABA is a
documented effective marker for compliance in chronic feeding trials. Analysis
of 24-h urine samples for PABA recovery will follow the method of Sharma et
al.[25]. Urea nitrogen and creatinine will be measured on a COBAS 400 Plus
analyzer. Urine samples will be collected at baseline to establish the PABA
composition of each individual's customary diet and again at weeks 4, 8 and
12. Creatinine will be measured to ensure complete 24-hr samples are
collected.Statistical Analysis: The analysis is a simple one-way ANOVA with
five treatment groups. Power calculations are based on the report of Raben et
al., [19]. We will recruit 140 individuals (28/group) to allow for attrition (double
our prior experience). We will have 80% power to detect differences in
average weight gain of 2.8 kgs.2. Evaluate the effects of aspartame on
appetite, gut peptides and glycemiaStudy design: Randomized, controlled
feeding trialSubjects:Males and females in roughly equal numbersAge 18-60
yearsBMI 18 - 25 kgm-2Not taking medications that affect metabolism or
appetiteNon- or low-user of low calorie sweeteners, including aspartameNo
reported aspartame sensitivityFasting blood glucose between 4.0 and 6.0
mmol/L (72 to 108 mg/dL) via capillary finger-stickIntervention: fruit-flavored
beverage containing aspartame or combination of beverage and
capsuleIntervention Dose: Three arms will be tested:0% (water)5 mg/kg
aspartame in a beverage15mg/kg aspartame, 5mg/kg as a beverage and the
balance in capsules.The beverage powder will be comprised of aspartame,
PABA and Kool-Aid. The capsules will contain the same dry ingredients as the
beverage. Each capsule will weigh 1 gram. Four doses will be prepared and
administered to participants according to their closest body weight.Statistical
Analysis: Study 2 is designed to measure the effects of aspartame ingestion
on appetite, gut peptide secretion and post-prandial glycemia. Data will be
analyzed by a mixed-model repeated measures analysis of variance.

PROGRESS: 2016/10 TO 2019/10
Target Audience:Our target audience is consumers, clinicians, policy makers
and scientists. Changes/Problems:Dr. Richard Mattes has taken on an
admistrative role as the Head of the Department of Public Health. What
opportunities for training and professional development has the project
provided?A good portion of the work was completed by graduate students.
They conducted the clinical procedures, analyzed biological samples, coded
and analyzed data, wrote reports andspoke at professional meetings. Thus,
they obtained considerable professional experience. How have the results
been disseminated to communities of interest?The findings from our work
have been distributed primarily through published journal articles, but also in
lay publications, and through presentations atprofessional meetings.
Interviews were also conducted with journalists from the lay community. What
do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported

IMPACT: 2016/10 TO 2019/10
What was accomplished under these goals? We published four manuscripts in
peer-reviewed journals. The basic findings were that most popular low calorie
sweeteners are not problematic for control of appetite, food intake, body
weight or glycaemia. Sucralose was actually beneficial for weight
management and saccharin was problematic. These findings raise questions
about recommending all low calorie sweeteners equally. They may have
discrepant physiological and behavioral effects. This has implications for
setting policy related to these products. The principle investigator also gave
six invited presentations at professional meetings.
PUBLICATIONS (not previously reported): 2016/10 TO 2019/10
1. Type: Journal Articles Status: Published Year Published: 2019 Citation: Am
J Clin Nutr 2019;109:1288-1301
2. Type: Journal Articles Status: Published Year Published: 2018 Citation:
Nutrition 2018;55-56:S6-S7
3. Type: Journal Articles Status: Published Year Published: 2019 Citation:
Current Opinion in Endocrine and Metabolic Research 2019;4:14-20
4. Type: Journal Articles Status: Published Year Published: 2018 Citation: J
Nutr 2018, doi: https://doi.org/10.1093/jn/nxy021