To Determine the Metabolomics and Microbiome Changes After Cranberry Juice Consumption Among Young Women
| Status: | Completed |
|---|---|
| Conditions: | Healthy Studies |
| Therapuetic Areas: | Other |
| Healthy: | No |
| Age Range: | 18 - 29 |
| Updated: | 11/18/2018 |
| Start Date: | January 16, 2017 |
| End Date: | November 15, 2018 |
The compliance of participants in cranberry related clinical trial is poor due to the
bitterness and astringency of cranberry products. There is a pressing need for an effective
approach to verify participant compliance. The PLS-DA (Partial Least Squares Discriminant
Analysis) /OPLS-DA (Orthogonal Partial Least Squares Discriminant Analysis) models
established from the investigator's previous study provide an opportunity to identify
cranberry juice consumers. In this blinded, placebo-controlled, cross-over study,
investigators hypothesize that the established statistical models of PLS-DA (Partial least
squares discriminant analysis) /OPLS-DA (Orthogonal partial least squares discriminant
analysis) can identify cranberry juice consumers and non-consumers by analyzing the
metabolomic changes in blood and urine among young women. In order to further explore the
function of cranberry, microbiome profile will also be compared between cranberry juice
consumption and placebo consumption after long-term treatment. Another comparison of
metabolomic changes between short-term treatment and long-term treatment will be made to
further identify candidate biomarkers of cranberry consumption and explore the correlation
between acute effects and chronic effects of cranberry.
bitterness and astringency of cranberry products. There is a pressing need for an effective
approach to verify participant compliance. The PLS-DA (Partial Least Squares Discriminant
Analysis) /OPLS-DA (Orthogonal Partial Least Squares Discriminant Analysis) models
established from the investigator's previous study provide an opportunity to identify
cranberry juice consumers. In this blinded, placebo-controlled, cross-over study,
investigators hypothesize that the established statistical models of PLS-DA (Partial least
squares discriminant analysis) /OPLS-DA (Orthogonal partial least squares discriminant
analysis) can identify cranberry juice consumers and non-consumers by analyzing the
metabolomic changes in blood and urine among young women. In order to further explore the
function of cranberry, microbiome profile will also be compared between cranberry juice
consumption and placebo consumption after long-term treatment. Another comparison of
metabolomic changes between short-term treatment and long-term treatment will be made to
further identify candidate biomarkers of cranberry consumption and explore the correlation
between acute effects and chronic effects of cranberry.
Consumption of cranberry juice has been related to the prevention of urinary tract infection.
However, due to the bitterness and astringency of cranberry products, the compliance of
participants in cranberry related clinical trial is poor, which makes it difficult to further
study the function of cranberry juice. Accordingly, there is a pressing need for an effective
approach to verify participant compliance. In the previous study, plasma and urinary
metabolome in young women following cranberry juice consumption have been investigated using
a global UHPLC (Ultra high-performance liquid chromatography) -Q-Orbitrap-HRMS
(High-resolution mass spectrometry) approach. PLS-DA (Partial least squares discriminant
analysis) analyses showed that both plasma and urinary metabolome in young women were altered
following cranberry consumption compared to baseline and apple juice consumption. Cranberry
juice consumption caused a greater excretion of both exogenous and endogenous metabolites.
These metabolites have been identified as candidate biomarkers for cranberry juice
consumption. The established PLS-DA (Partial least squares discriminant analysis) /OPLS-DA
(Orthogonal partial least squares discriminant analysis) models provide an opportunity to
identify cranberry juice consumers.
It is unknown whether cranberry juice consumption influences gut microbiota profile in human.
But several studies suggested that a polyphenol-rich diet improves gut microbiota profile.
The previous in vitro study with human gut microbiota indicated that microbial catabolism of
A-type cranberry procyanidins produced metabolites with different profiles and structures
compared with those from B-type procyanidins.
Understanding microbiome change in human gut will enable the investigators to correlate and
explain metabolome changes in the urine and blood, and also will explain the health benefits
of cranberries.
With all the samples will be collected in this study, the metabolomics changes between
short-term cranberry juice treatment (3days) and long-term treatment (21days) can be
compared. Since the urinary tract protection effects of cranberry juices are often associated
with chronic consumption, more metabolomic changes are expected to be identified by a
long-term study.
Research Plan:
Specific aim 1: Twenty (20) healthy female college students aged 18-29 with normal BMI (Body
mass index) (18.5-25) and at least 110 pounds in weight will be recruited on UF (University
of Florida) campus.
Advertisement will be in the form of flyers. Contact information of investigators will be
included in the flyer. Questionnaire will be used to obtain information about subjects for
prescreen. All participants will be received written and oral information regarding the
natural and potential risks of the study.
Participants will be advised to avoid proanthocyanidins-rich foods including cranberries,
blueberries, strawberries plum, raspberries, apple, grapes, grape seed extract, pycnogenol,
red wine, tea, chocolate and other cocoa products or other cranberry based supplements from
the beginning of 10-day (1st-9th ) run in period to the end of the study. Participants will
be suggested to consume orange, watermelon, honey dew, cantaloupe for replacement. Urine and
stool sample collection kits will be distributed to all participants and participants will be
trained to use the collection kits. One fecal sample will be collected by participants during
any day from the 7th to 9th day. On the 10th day morning, overnight fast urine sample will be
self-collected and subjects will have blood sample drawn (2 tubes, 30mL total). Breakfast
will be provided for participants after fasting blood sample collection. A trained and
certified phlebotomist will be hired to draw blood. After blood drawn, each participant will
consume 1 bottle (250mL) of cranberry or placebo juice and another 5 bottles of cranberry or
placebo juice will be given to each subject to consume in the evening of the 10th day and in
the morning and evening of the 11th and12th day. One fecal sample will also be collected
during the 12th- 4th day. All subjects will return to the clinical unit in the morning of the
13th day with a self-collected overnight fast urine sample and blood samples will be
collected 30-60min after participants consume 1 bottle of cranberry or placebo juice.
Cranberry or placebo juice will be distributed to each of the subjects to consume in the
evening of the 13th day and in the morning and evening during the 14th- 29th day. Appointment
will be scheduled weekly or upon the request of participants during the 14th to 29th day. One
fecal sample will be collected during the 29th to 31st day. In the morning of the 30th day,
overnight fast urine sample will be self-collected and subjects will have blood sample drawn
after consume cranberry juice or placebo. After 14-days washout period, participants will
switch to the alternative regimen and repeat the protocol. All samples collected will be
aliquoted and kept in -80°C. It's unlikely to happen, but if any subject has any serious
adverse effect after juice consumption, participant will be removed from the study depending
on her situation for safety concern.
Preparation of urine and plasma sample collected at the13th, 48th day (3 days later after
cranberry juice/placebo consumption) will follow established protocols. LC-HRMS (Liquid
chromatography - High-resolution mass spectrometry) data will be normalized and imported to
SIMCA (Version 13.0.3, Umetrics, Umea, Sweden) to use previously established PLS-DA (Partial
least squares discriminant analysis) /OPLS-DA (Orthogonal partial least squares discriminant
analysis) models to blindly identify cranberry juice consumers. Results will be sent to Ocean
Spray for verification.
Specific aim 2:
The baseline fecal samples and fecal sample collected around the 30th, 65th day (21 days
later after cranberry juice/placebo consumption) will be analyzed for microbiome composition.
Genomic microbial DNA will be extracted from stool samples using MoBlo Power soil DNA
isolation kits.. Microbiota change in the gut will be correlated with metabolome changes in
the urine using redundancy and discriminant analysis. Fecal mucin and IgA (Immunoglobulin A,
indicators of intestinal barrier), short chain fatty acids and ammonia in the stool samples
will be analyzed using specific assay kits or HPLC (High-performance liquid chromatography)
method.
Specific aim 3:
Preparation of urine and plasma sample collected at the 30th, 65th day (21 days later after
cranberry juice/placebo consumption) will follow established protocols. UHPLC (Ultra
high-performance liquid chromatography) -Q-Orbitrap-HRMS (High-resolution mass spectrometry)
analyses, Multivariate data processing and statistical analysis will be the same as the
experiment process discussed in Specific aim 2.
However, due to the bitterness and astringency of cranberry products, the compliance of
participants in cranberry related clinical trial is poor, which makes it difficult to further
study the function of cranberry juice. Accordingly, there is a pressing need for an effective
approach to verify participant compliance. In the previous study, plasma and urinary
metabolome in young women following cranberry juice consumption have been investigated using
a global UHPLC (Ultra high-performance liquid chromatography) -Q-Orbitrap-HRMS
(High-resolution mass spectrometry) approach. PLS-DA (Partial least squares discriminant
analysis) analyses showed that both plasma and urinary metabolome in young women were altered
following cranberry consumption compared to baseline and apple juice consumption. Cranberry
juice consumption caused a greater excretion of both exogenous and endogenous metabolites.
These metabolites have been identified as candidate biomarkers for cranberry juice
consumption. The established PLS-DA (Partial least squares discriminant analysis) /OPLS-DA
(Orthogonal partial least squares discriminant analysis) models provide an opportunity to
identify cranberry juice consumers.
It is unknown whether cranberry juice consumption influences gut microbiota profile in human.
But several studies suggested that a polyphenol-rich diet improves gut microbiota profile.
The previous in vitro study with human gut microbiota indicated that microbial catabolism of
A-type cranberry procyanidins produced metabolites with different profiles and structures
compared with those from B-type procyanidins.
Understanding microbiome change in human gut will enable the investigators to correlate and
explain metabolome changes in the urine and blood, and also will explain the health benefits
of cranberries.
With all the samples will be collected in this study, the metabolomics changes between
short-term cranberry juice treatment (3days) and long-term treatment (21days) can be
compared. Since the urinary tract protection effects of cranberry juices are often associated
with chronic consumption, more metabolomic changes are expected to be identified by a
long-term study.
Research Plan:
Specific aim 1: Twenty (20) healthy female college students aged 18-29 with normal BMI (Body
mass index) (18.5-25) and at least 110 pounds in weight will be recruited on UF (University
of Florida) campus.
Advertisement will be in the form of flyers. Contact information of investigators will be
included in the flyer. Questionnaire will be used to obtain information about subjects for
prescreen. All participants will be received written and oral information regarding the
natural and potential risks of the study.
Participants will be advised to avoid proanthocyanidins-rich foods including cranberries,
blueberries, strawberries plum, raspberries, apple, grapes, grape seed extract, pycnogenol,
red wine, tea, chocolate and other cocoa products or other cranberry based supplements from
the beginning of 10-day (1st-9th ) run in period to the end of the study. Participants will
be suggested to consume orange, watermelon, honey dew, cantaloupe for replacement. Urine and
stool sample collection kits will be distributed to all participants and participants will be
trained to use the collection kits. One fecal sample will be collected by participants during
any day from the 7th to 9th day. On the 10th day morning, overnight fast urine sample will be
self-collected and subjects will have blood sample drawn (2 tubes, 30mL total). Breakfast
will be provided for participants after fasting blood sample collection. A trained and
certified phlebotomist will be hired to draw blood. After blood drawn, each participant will
consume 1 bottle (250mL) of cranberry or placebo juice and another 5 bottles of cranberry or
placebo juice will be given to each subject to consume in the evening of the 10th day and in
the morning and evening of the 11th and12th day. One fecal sample will also be collected
during the 12th- 4th day. All subjects will return to the clinical unit in the morning of the
13th day with a self-collected overnight fast urine sample and blood samples will be
collected 30-60min after participants consume 1 bottle of cranberry or placebo juice.
Cranberry or placebo juice will be distributed to each of the subjects to consume in the
evening of the 13th day and in the morning and evening during the 14th- 29th day. Appointment
will be scheduled weekly or upon the request of participants during the 14th to 29th day. One
fecal sample will be collected during the 29th to 31st day. In the morning of the 30th day,
overnight fast urine sample will be self-collected and subjects will have blood sample drawn
after consume cranberry juice or placebo. After 14-days washout period, participants will
switch to the alternative regimen and repeat the protocol. All samples collected will be
aliquoted and kept in -80°C. It's unlikely to happen, but if any subject has any serious
adverse effect after juice consumption, participant will be removed from the study depending
on her situation for safety concern.
Preparation of urine and plasma sample collected at the13th, 48th day (3 days later after
cranberry juice/placebo consumption) will follow established protocols. LC-HRMS (Liquid
chromatography - High-resolution mass spectrometry) data will be normalized and imported to
SIMCA (Version 13.0.3, Umetrics, Umea, Sweden) to use previously established PLS-DA (Partial
least squares discriminant analysis) /OPLS-DA (Orthogonal partial least squares discriminant
analysis) models to blindly identify cranberry juice consumers. Results will be sent to Ocean
Spray for verification.
Specific aim 2:
The baseline fecal samples and fecal sample collected around the 30th, 65th day (21 days
later after cranberry juice/placebo consumption) will be analyzed for microbiome composition.
Genomic microbial DNA will be extracted from stool samples using MoBlo Power soil DNA
isolation kits.. Microbiota change in the gut will be correlated with metabolome changes in
the urine using redundancy and discriminant analysis. Fecal mucin and IgA (Immunoglobulin A,
indicators of intestinal barrier), short chain fatty acids and ammonia in the stool samples
will be analyzed using specific assay kits or HPLC (High-performance liquid chromatography)
method.
Specific aim 3:
Preparation of urine and plasma sample collected at the 30th, 65th day (21 days later after
cranberry juice/placebo consumption) will follow established protocols. UHPLC (Ultra
high-performance liquid chromatography) -Q-Orbitrap-HRMS (High-resolution mass spectrometry)
analyses, Multivariate data processing and statistical analysis will be the same as the
experiment process discussed in Specific aim 2.
Inclusion Criteria:
- Healthy
- Normal BMI (18.5-25)
- Body weight ≥110 pounds
Exclusion Criteria:
- Gastrointestinal disorders
- Urological diseases
- Metabolic disorders
- Smokers
- Pregnancy
- Previous use of antibiotics or probiotics.
We found this trial at
1
site
Gainesville, Florida 32611
Phone: 352-294-3731
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