Healthy
Conditions
Brief summary
People who eat diets low in choline should deplete their choline (Cho) stores, and measurements of Cho pool size using isotope dilution should reflect this depletion. Investigators will identify a biomarker panel that correlates well with measured Cho pool size across the range of different degrees of depletion.The investigators propose that, as body stores of Cho diminish, cells and organs will reach the point when metabolism/function in the cell is altered, and that this will result in a progression of changes in biomarkers that reflect Cho status.
Detailed description
Choline (Cho) is an essential nutrient and most Americans' diets do not achieve the recommended intake. Diets low in Cho are associated with liver and muscle disease and with suboptimal fetal development, while diets too high in choline may be associated with increased risk for heart disease. Cho is a required nutrient and in 1998, an Adequate Intake (AI) and a Tolerable Upper Limit (UL) for Cho was established In 2016, the US Food and Drug Administration (FDA) set a Recommended Daily Intake (RDI) for Cho based on the AIs as part of the new Nutrition Facts label for packaged foods (published in the Federal Register on May 27, 2016; FDA-2012-N-1210-0875, Federal Register Number:2016-11867). The AI/RDI varies by age and gender, but is 550 mg/d in adolescent and adult men and 425 mg/d in adult women (more in pregnant and lactating women) and 400 mg/day for adolescent women. There is no validated biomarker for choline status (the availability of the various forms of Cho needed to sustain optimal cellular function). Measurement of plasma Cho concentrations is not adequate as plasma choline is homeostatically regulated. Based on extensive preliminary and published data this group identified a panel of potential biomarkers that could be used to assess Cho status, and now the investigators propose to validate this biomarker panel against measures of Cho pool size using isotope dilution. The largest stores of Cho are located in the liver, and mass resonance spectroscopy (MRS) of liver has been used in the past to assess Cho status in humans. This method is not practical for use as a biomarker in clinical or public health practice as it is expensive and the availability of the instrumentation is limited. However, the MRS can be utilized to confirm correlations between the biomarker panel and the isotope dilution method. Liver biopsy is risky and not practical, making measurement of hepatic Cho and Cho metabolites concentrations a poor choice for assessing Cho status. Perhaps there is a panel of biomarkers that together will more accurately and reliably reflect Cho status. By making measurements in people fed 3 different dietary amounts of Cho for two weeks at a time, and comparing the biomarker measures to body total Cho pool size assessed using isotope dilution (a proxy for the availability of the various forms of Cho), investigators will be able to identify the combination of biomarkers and algorithm for calculating a Cho status score that best predicts total Cho pool size, and therefore predicts choline nutritional status (the availability of the various forms of Cho needed to sustain cellular function). People who eat diets low in choline should deplete their choline (Cho) stores, and measurement of Cho pool size using isotope dilution should reflect this depletion. The investigators will identify a biomarker panel that correlates well with measured Cho pool size across the range of different degrees of depletion. This study tests a method for using stable isotope dilution to measure body choline stores, and then asks how this measure correlates with a panel of biomarkers in plasma and with liver fat measured using Fibroscan®. Using isotope dilution can provide an estimate of the size of the body pool of Cho. The investigators' proposed method is conceptually similar to the method for measuring total body water from a bolus dose of labeled water. Similar methodology was used recently in studies of metabolic flux of Cho in pregnant women. Isotope dilution is a well-established method used to estimate pool size for other nutrients, such as vitamin A. Similar to vitamin A, the major storage pools for Cho are in the liver, and ingested Cho is rapidly absorbed and accumulated by liver. MRS/MRI scans will also be performed to investigate correlation between these gold standard measures and the other methods described above. Participants will consume meals provided in two week dietary intervals with 3 different levels of choline with a 2 week washout periods between those dietary intervals. Participants will receive 100% of the recommended intake (RDI) of Cho (550mg Cho/day); 50% of the RDI of Cho (275mg/day); and 25% of the RDI of Cho (137.5mg/day). The meal order will be randomly assigned and all participants will receive all diets at some point in the study. There will be a minimum of a two week washout between diet intervals. Both participants and researchers will be blinded to the diet order. Participants will have brief exercise challenges (Biodex) to assess muscle function as an additional predictor of choline status. Participants enrolled prior to 3/2020 completed MRI/MRS scans. We have determined that Fibroscan provides adequate measurement of liver fat such that we eliminated the added inconvenience to participants of travel to Winston-Salem for MR scanning. Saliva, stool, and urine samples will be collected.
Interventions
OTHER25% Cho diet
Subjects will consume meals containing 25% of recommended intake of Choline for 2 weeks. On day 12 of the diet period, subjects will consume 250 mg of Cho in the form of Cho chloride (trimethyl-d9, 9%), Cambridge Isotope Laboratories, Tewksbury, Massachusetts, (USA), as a bolus.
OTHER50% Cho diet
Subjects will consume meals containing 50% of recommended intake of Choline for 2 weeks. On day 12 of the diet period, subjects will consume 250 mg of Cho in the form of Cho chloride (trimethyl-d9, 9%), Cambridge Isotope Laboratories, Tewksbury, Massachusetts, (USA), as a bolus.
OTHER100% Cho diet
Subjects will consume meals containing 100% of recommended intake of Choline for 2 weeks. On day 12 of the diet period, subjects will consume 250 mg of Cho in the form of Cho chloride (trimethyl-d9, 9%), Cambridge Isotope Laboratories, Tewksbury, Massachusetts, (USA), as a bolus.
Sponsors
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
University of North Carolina, Chapel Hill
Study design
Allocation
RANDOMIZED
Intervention model
CROSSOVER
Primary purpose
SCREENING
Masking
TRIPLE (Subject, Investigator, Outcomes Assessor)
Masking description
Participant dietary arm assignment is randomized by a randomization plan created by the study coordinator at www.randomization.com. Each of the 3 hormonal related demographic groups (male, premenopausal females, and postmenopausal females) will have a list of the same order of diets created by the randomizer. Each participant will be assigned upon entry into the study into the next open diet for their group as ordered by the randomizer. No one collecting or processing data will be informed of the choline levels the participant is experiencing in their dietary arms at any given time to attempt to eliminate bias and ensure appropriate data collection. All staff who interact with participants or who are handling samples/data, will not be informed of the code linking dietary choline levels to diet order.
Intervention model description
Healthy volunteers will consume meals, provided by the investigator, in two week intervals with 3 different levels of choline (Cho). Participants will receive 100% of the recommended daily intake (RDI) of Cho (550 mg Cho/day); 50% of the RDI of Cho (275mg/day); and 25% of the RDI of Cho (137.5 mg/day). The meal order will be randomly assigned and all participants will receive all diets at some point in the study. There will be a minimum of a two week washout between diet intervals where participants return to their regular diets. Both participants and researchers will be blinded to the diet order.
Eligibility
Sex/Gender
ALL
Age
17 Years to 70 Years
Healthy volunteers
Yes
Inclusion criteria
* Provision of signed and dated informed consent form * Weigh 130-177lbs (or if over 177 must have BMI under 28) * Stated willingness to comply with all study procedures and availability for the duration of the study * Male or female, aged 17-70 years * In good general health as evidenced by medical history, clinical chemistries, physical exam, and BMI≤ 30 or if over 177lbs with a BMI under 28 * Women who are included in the study and are of pregnancy potential will have a urine pregnancy test at the beginning and end of each dietary intervention arm and must be using some form of birth control during the study.
Exclusion criteria
* using drugs or medication known to be damaging to liver or muscle at typically prescribed doses or that have the potential to alter Cho metabolism (e.g., methotrexate); * history of hepatic, renal, or other chronic systemic disease. * subjects with liver abnormalities (e.g.cysts) as determined by ultrasound * current smokers * consume \>2 alcoholic beverages/d or \>14/wk * substance abusers or drug addicted * eating unusual diet that would interfere with the study * food allergies, (e.g., soy) or any problems with eating all foods on required study diet * using Cho-containing dietary supplements * women who are breastfeeding, pregnant, or plan to become pregnant due to potential risk to fetus/child of low choline diet * performing intense exercise of more than 1 hour a day or other intense muscle building exercise (such as weightlifting beyond low weight repetitions) * Actively participating in other research study where required to exercise or ingest any food, medicine, or supplement in any manner * have been screened for this study between August and March and have not provided proof of flu vaccination prior to enrollment
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Change in Liver Fat Content Based on CAP Values | Day 1 and Day 15 of respective Cho diet | Controlled attenuation parameter (CAP) as measured by Fibroscan is an ultrasound-based technique to measure liver fat. This method will be used with other biomarkers to indicate functional signs of choline status. |
| Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | At the end of 2 weeks of respective Cho diet | Metabolomics was conducted on plasma that was collected from individuals at the end of each 2-week diet period. UHPLC High Resolution Mass Spectrometry was used for differential profiling (PMID: 33415121). Supervised Orthogonal Partial Least Squares Discriminant Analysis was used to determine signals that differentiated the 25% choline dietary group from the 100% choline dietary group. Metabolites that differentiated the 25% and 100% choline dietary groups with variable importance to projection (VIP) \>1 and p-value \< 0.05 are reported. The signals for these metabolites were matched by retention time, exact mass, and MS/MS to standards run on the same platform. Because this is a differential profiling method (not quantitative), the mean and standard deviation of peak intensities detected on the untargeted platform are reported. Results are reported for the selected metabolites for the 25%, 50%, and 100% choline dietary groups. Ratios can be obtained by division of the intensity data. |
| Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | 24 hours following administration of choline-d9 on day 12 of respective dietary intervention | DNA was collected and evaluated for the presence of the PEMT SNP rs12325817. Genotypes was measured by real time polymerase chain reaction (RT-PCR). The magnitude of changes in choline pool size as measured by IER at the end of each dietary intervention was compared among subjects with different genotypes in the PEMT SNP. Linear mixed model with repeated measures was performed for each group (healthy males, pre- and postmenopausal females) separately to study the genotype effect and genotype x diet interaction effect on choline pool size. |
| Ratio of Liver Choline Pool Size by Isotope Dilution | 24 hours following administration of choline-d9 on day 12 of respective dietary intervention | The liver choline pool determined by the dilution of the deuterated choline metabolites formed in liver and released to plasma as measured by isotopic enrichment ratio (IER). The IER for a given metabolite is defined as the concentration of a deuterated metabolite divided by the sum of deuterated and non-deuterated metabolite. |
| Difference in Choline Deficiency Signature | At the end of 2 weeks of respective Cho diet | Plasma choline metabolites (micromolar): choline, dimethylglycine, betaine, phosphatidylcholine, sphingomyelin, trimethylamine-oxide, and homocysteine measured by targeted metabolomic profiling. The signature for choline deficiency is defined by choline \<0.793 mmol/L or betaine \<34.9 mmol/L. The levels of these metabolites at the end of each intervention will be compared. The association between choline metabolites and choline pool size will be investigated. |
| Comparison of Choline Pool Size Between Participants With and Without Choline Metabolites Signature During Cho Depletion | 24 hours following administration of choline-d9 on day 12 of 25% Cho diet | The 25% Cho arm was selected because only at that intake level is sufficient depletion achieved. Participants with plasma choline \<0.793 mmol/L or betaine \<34.9 mmol/L were considered as choline depleted (showing signature), participants with plasma choline \>=0.793 mmol/L and betaine \>=34.9 mmol/L were considered as not choline depleted (not showing signature). Available choline pool size was determined by the dilution of the deuterated choline metabolites formed in liver and released to plasma as measured by isotopic enrichment ratio (IER). The IER for a given metabolite is defined as the concentration of a deuterated metabolite divided by the sum of deuterated and non-deuterated metabolite. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Validation of Isotope Dilution Method to Assess Choline Pool Size by Magnetic Resonance Spectroscopy (MRS) | At the end of 2 weeks of respective Cho diet | MRS is a direct measurement of liver choline content. Changes in liver choline by MRS should correlate with changes in liver choline inferred by calculation of isotope enrichment ratio (IER) of plasma metabolites. Pearson correlation coefficient used to study the correlation between data generated from the two types of measurements. |
Other
| Measure | Time frame | Description |
|---|---|---|
| SNPs That Create Inefficiencies in Choline Metabolism Associated With Change in Choline Pool Size and Choline Biomarkers | At baseline visit | Exploratory analysis of \>2 million SNPs measured on a custom Illumina Expanded Multi-Ethnic genotyping array (Mega-Ex). The same analysis described for Outcome 4 will be applied for Outcome 7. Benjamini-Hochberg method for False Discovery Rate (FDR) correction will be used for multiple testing correction. |
Countries
United States
Participant flow
Participants by arm
| Arm | Count |
|---|---|
| Total Number of Participants All participants who were randomly assigned to diet sequence containing 137.5 mg (25% Cho diet), 275 mg (50% Cho diet), and 550 mg (100% Cho diet) for two weeks each with 2 weeks of washout between. | 101 |
| Total | 101 |
Baseline characteristics
| Characteristic | Total Number of Participants |
|---|---|
| Age, Categorical <=18 years | 1 Participants |
| Age, Categorical >=65 years | 0 Participants |
| Age, Categorical Between 18 and 65 years | 100 Participants |
| Ethnicity (NIH/OMB) Hispanic or Latino | 9 Participants |
| Ethnicity (NIH/OMB) Not Hispanic or Latino | 92 Participants |
| Ethnicity (NIH/OMB) Unknown or Not Reported | 0 Participants |
| Menopausal Status Post-Menopausal Females | 21 Participants |
| Menopausal Status Pre-Menopausal Females | 50 Participants |
| Race (NIH/OMB) American Indian or Alaska Native | 0 Participants |
| Race (NIH/OMB) Asian | 2 Participants |
| Race (NIH/OMB) Black or African American | 13 Participants |
| Race (NIH/OMB) More than one race | 1 Participants |
| Race (NIH/OMB) Native Hawaiian or Other Pacific Islander | 0 Participants |
| Race (NIH/OMB) Unknown or Not Reported | 0 Participants |
| Race (NIH/OMB) White | 85 Participants |
| Region of Enrollment United States | 101 Participants |
| Sex: Female, Male Female | 71 Participants |
| Sex: Female, Male Male | 30 Participants |
Adverse events
| Event type | EG000 affected / at risk | EG001 affected / at risk | EG002 affected / at risk |
|---|---|---|---|
| deaths Total, all-cause mortality | 0 / 91 | 0 / 84 | 0 / 86 |
| other Total, other adverse events | 0 / 91 | 0 / 84 | 0 / 86 |
| serious Total, serious adverse events | 0 / 91 | 0 / 84 | 0 / 86 |
Outcome results
Primary
Change in Liver Fat Content Based on CAP Values
Controlled attenuation parameter (CAP) as measured by Fibroscan is an ultrasound-based technique to measure liver fat. This method will be used with other biomarkers to indicate functional signs of choline status.
Time frame: Day 1 and Day 15 of respective Cho diet
Population: Participants who completed at least 2 arms were included in the analysis. Out of 101 randomized, 78 met this criteria. Of the 78, 3 were excluded for noncompliance, leaving 75 participants. Of these 75, 75 completed the 25%, 73 completed the 50% \& 75 completed the 100% diets. Fibroscan data were missing for 2 participants in the 25% Cho arm and 2 in the 50% arm.
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| 25% Cho | Change in Liver Fat Content Based on CAP Values | 0.0208 dB/m | Standard Deviation 0.1513 |
| 50% Cho | Change in Liver Fat Content Based on CAP Values | 0.6958 dB/m | Standard Deviation 0.2144 |
| 100% Cho | Change in Liver Fat Content Based on CAP Values | 0.0135 dB/m | Standard Deviation 0.1482 |
p-value: 0.0842Mixed Models Analysis
p-value: 0.0819Wilcoxon (Mann-Whitney)
p-value: 0.9015t-test, 2 sided
p-value: 0.1191Wilcoxon (Mann-Whitney)
Primary
Comparison of Choline Pool Size Between Participants With and Without Choline Metabolites Signature During Cho Depletion
The 25% Cho arm was selected because only at that intake level is sufficient depletion achieved. Participants with plasma choline \<0.793 mmol/L or betaine \<34.9 mmol/L were considered as choline depleted (showing signature), participants with plasma choline \>=0.793 mmol/L and betaine \>=34.9 mmol/L were considered as not choline depleted (not showing signature). Available choline pool size was determined by the dilution of the deuterated choline metabolites formed in liver and released to plasma as measured by isotopic enrichment ratio (IER). The IER for a given metabolite is defined as the concentration of a deuterated metabolite divided by the sum of deuterated and non-deuterated metabolite.
Time frame: 24 hours following administration of choline-d9 on day 12 of 25% Cho diet
Population: Participants who completed at least 2 arms were included in the analysis. Out of 101 randomized, 78 met this criteria. Of the 78, 3 were excluded for noncompliance, leaving 75 participants. All 75 participants completed 25% Cho diet. Only data from the 25% Cho diet (choline depleted state) were analyzed.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| 25% Cho | Comparison of Choline Pool Size Between Participants With and Without Choline Metabolites Signature During Cho Depletion | Betaine IER | 0.0234 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0087 |
| 25% Cho | Comparison of Choline Pool Size Between Participants With and Without Choline Metabolites Signature During Cho Depletion | Choline IER | -0.0005 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0049 |
| 25% Cho | Comparison of Choline Pool Size Between Participants With and Without Choline Metabolites Signature During Cho Depletion | Phosphatidylcholine IER | 0.0115 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0034 |
| 50% Cho | Comparison of Choline Pool Size Between Participants With and Without Choline Metabolites Signature During Cho Depletion | Phosphatidylcholine IER | 0.0118 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.003 |
| 50% Cho | Comparison of Choline Pool Size Between Participants With and Without Choline Metabolites Signature During Cho Depletion | Betaine IER | 0.0241 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0057 |
| 50% Cho | Comparison of Choline Pool Size Between Participants With and Without Choline Metabolites Signature During Cho Depletion | Choline IER | 0.0025 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0051 |
Comparison: Betaine IERp-value: 0.6985t-test, 2 sided
Comparison: Choline IERp-value: 0.035t-test, 2 sided
Comparison: Phosphatidylcholine IERp-value: 0.6864t-test, 2 sided
Primary
Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817)
DNA was collected and evaluated for the presence of the PEMT SNP rs12325817. Genotypes was measured by real time polymerase chain reaction (RT-PCR). The magnitude of changes in choline pool size as measured by IER at the end of each dietary intervention was compared among subjects with different genotypes in the PEMT SNP. Linear mixed model with repeated measures was performed for each group (healthy males, pre- and postmenopausal females) separately to study the genotype effect and genotype x diet interaction effect on choline pool size.
Time frame: 24 hours following administration of choline-d9 on day 12 of respective dietary intervention
Population: Participants who completed at least 2 arms were included in the analysis. Out of 101 randomized, 78 met this criteria. Of the 78, 3 were excluded for noncompliance. Of the 75 remaining, one had missing SNP data. One had missing isotope dilution data in 50% Cho. Two subjects did not complete 50% Cho. That brings the sample size down to 74, 71, and 74 for the 25%, 50%, and 100%, respectively. Overall Number of Participants Analyzed includes Pre- and Post-Menopausal Females \& Males.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| 25% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Betaine IER | 0.0264 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0059 |
| 25% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Choline IER | -0.0018 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0051 |
| 25% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Choline IER | 0.0043 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0026 |
| 25% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Phosphatidylcholine IER | 0.0095 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0021 |
| 25% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Choline IER | 0.0014 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0048 |
| 25% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Betaine IER | 0.0212 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.007 |
| 25% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Phosphatidylcholine IER | 0.0131 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0002 |
| 25% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Betaine IER | 0.0259 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0101 |
| 25% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Phosphatidylcholine IER | 0.0113 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.003 |
| 50% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Choline IER | -0.0018 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0063 |
| 50% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Betaine IER | 0.025 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0053 |
| 50% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Phosphatidylcholine IER | 0.011 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0044 |
| 50% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Phosphatidylcholine IER | 0.0135 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0021 |
| 50% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Betaine IER | 0.02 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0033 |
| 50% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Betaine IER | 0.0201 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.01 |
| 50% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Choline IER | -0.0001 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0039 |
| 50% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Phosphatidylcholine IER | 0.009 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0014 |
| 50% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Choline IER | 0.0029 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0034 |
| 100% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Betaine IER | 0.0261 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0064 |
| 100% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Choline IER | 0.0017 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.005 |
| 100% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Betaine IER | 0.0265 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0124 |
| 100% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Phosphatidylcholine IER | 0.011 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0024 |
| 100% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Phosphatidylcholine IER | 0.013 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0027 |
| 100% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Choline IER | -0.0011 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0028 |
| 100% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Betaine IER | 0.024 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0045 |
| 100% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Phosphatidylcholine IER | 0.0127 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0043 |
| 100% Cho | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Choline IER | 0.003 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0061 |
| 50% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Phosphatidylcholine IER | 0.0184 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0034 |
| 50% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Phosphatidylcholine IER | 0.011 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0027 |
| 50% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Phosphatidylcholine IER | 0.0071 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0049 |
| 50% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Betaine IER | 0.0136 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0126 |
| 50% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Betaine IER | 0.0244 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0073 |
| 50% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Choline IER | 0.0006 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0084 |
| 50% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Choline IER | 0.0097 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0022 |
| 50% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Betaine IER | 0.0419 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0264 |
| 50% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Choline IER | -0.0004 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0051 |
| 50% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Betaine IER | 0.0241 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0045 |
| 50% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Phosphatidylcholine IER | 0.01 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.001 |
| 50% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Phosphatidylcholine IER | 0.011 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0032 |
| 50% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Choline IER | 0.0007 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0042 |
| 50% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Choline IER | 0.0004 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0054 |
| 50% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Betaine IER | 0.0224 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0059 |
| 50% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Phosphatidylcholine IER | 0.0111 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0019 |
| 50% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Betaine IER | 0.0212 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0026 |
| 50% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Choline IER | 0.0028 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0026 |
| 50% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Choline IER | -0.0007 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0036 |
| 50% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Phosphatidylcholine IER | 0.0115 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0033 |
| 50% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Betaine IER | 0.0262 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0073 |
| 50% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Phosphatidylcholine IER | 0.0121 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0041 |
| 50% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Phosphatidylcholine IER | 0.0123 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0016 |
| 50% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Choline IER | 0.0044 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0045 |
| 50% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Betaine IER | 0.0222 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0021 |
| 50% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Choline IER | 0.0029 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0045 |
| 50% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Betaine IER | 0.028 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0127 |
| 100% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Choline IER | 0.0004 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0044 |
| 100% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Betaine IER | 0.0243 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0072 |
| 100% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Phosphatidylcholine IER | 0.0107 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0024 |
| 100% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Betaine IER | 0.0232 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0037 |
| 100% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Choline IER | 0.0047 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0022 |
| 100% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Phosphatidylcholine IER | 0.0096 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0016 |
| 100% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Betaine IER | 0.0247 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0053 |
| 100% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Choline IER | 0.0061 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0005 |
| 100% Cho, rs12325817, CC | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Phosphatidylcholine IER | 0.0127 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0007 |
| 100% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Phosphatidylcholine IER | 0.0086 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0011 |
| 100% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Phosphatidylcholine IER | 0.0105 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0025 |
| 100% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Betaine IER | 0.0175 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0026 |
| 100% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Choline IER | 0.0008 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0042 |
| 100% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Phosphatidylcholine IER | 0.0094 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0035 |
| 100% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Betaine IER | 0.0213 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0046 |
| 100% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Choline IER | 0.0029 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0038 |
| 100% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Betaine IER | 0.0201 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0077 |
| 100% Cho, rs12325817, CG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Choline IER | 0.0019 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0027 |
| 100% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Betaine IER | 0.0213 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0036 |
| 100% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Choline IER | 0.0026 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0035 |
| 100% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Betaine IER | 0.02 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0086 |
| 100% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Choline IER | 0.0041 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0042 |
| 100% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Phosphatidylcholine IER | 0.01 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0017 |
| 100% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Choline IER | 0.0001 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0039 |
| 100% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Pre-Menopausal Females: Betaine IER | 0.0247 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0039 |
| 100% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Males: Phosphatidylcholine IER | 0.0112 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0012 |
| 100% Cho, rs12325817, GG | Comparison of Choline Pool Size Between Participants With Different Genotypes in Phosphatidylethanolamine-N-methyltransferase (PEMT) Single Nucleotide Polymorphism (SNP rs12325817) | Post-Menopausal Females: Phosphatidylcholine IER | 0.0096 isotopic enrichment ratio (uM/uM) | Standard Deviation 0.0023 |
Comparison: Diet x SNP interaction effect on Betaine IER in Pre-Menopausal Femalesp-value: 0.9567Mixed Models Analysis
Comparison: Diet x SNP interaction effect on Betaine IER in Post-Menopausal Femalesp-value: 0.0899Mixed Models Analysis
Comparison: Diet x SNP interaction effect on Betaine IER in Malesp-value: 0.9003Mixed Models Analysis
Comparison: Diet x SNP interaction effect on Choline IER in Pre-Menopausal Femalesp-value: 0.9932Mixed Models Analysis
Comparison: Diet x SNP interaction effect on Choline IER in Post-Menopausal Femalesp-value: 0.3984Mixed Models Analysis
Comparison: Diet x SNP interaction effect on Choline IER in Malesp-value: 0.308Mixed Models Analysis
Comparison: Diet x SNP interaction effect on Phosphatidylcholine IER in Pre-Menopausal Femalesp-value: 0.7603Mixed Models Analysis
Comparison: Diet x SNP interaction effect on Phosphatidylcholine IER in Post-Menopausal Femalesp-value: 0.2995Mixed Models Analysis
Comparison: Diet x SNP interaction effect on Phosphatidylcholine IER in Malesp-value: 0.0003Mixed Models Analysis
Comparison: SNP effect on Betaine IER in Pre-Menopausal Femalesp-value: 0.1303Mixed Models Analysis
Comparison: SNP effect on Betaine IER in Post-Menopausal Femalesp-value: 0.0015Mixed Models Analysis
Comparison: SNP effect on Betaine IER in Malesp-value: 0.4185Mixed Models Analysis
Comparison: SNP effect on Choline IER in Pre-Menopausal Femalesp-value: 0.3464Mixed Models Analysis
Comparison: SNP effect on Choline IER in Post-Menopausal Femalesp-value: 0.0019Mixed Models Analysis
Comparison: SNP effect on Choline IER in Malesp-value: 0.2091Mixed Models Analysis
Comparison: SNP effect on Phosphatidylcholine IER in Pre-Menopausal Femalesp-value: 0.8141Mixed Models Analysis
Comparison: SNP effect on Phosphatidylcholine IER in Post-Menopausal Femalesp-value: 0.0187Mixed Models Analysis
Comparison: SNP effect on Phosphatidylcholine IER in Malesp-value: 0.0107Mixed Models Analysis
Primary
Difference in Choline Deficiency Signature
Plasma choline metabolites (micromolar): choline, dimethylglycine, betaine, phosphatidylcholine, sphingomyelin, trimethylamine-oxide, and homocysteine measured by targeted metabolomic profiling. The signature for choline deficiency is defined by choline \<0.793 mmol/L or betaine \<34.9 mmol/L. The levels of these metabolites at the end of each intervention will be compared. The association between choline metabolites and choline pool size will be investigated.
Time frame: At the end of 2 weeks of respective Cho diet
Population: Participants who completed at least 2 arms were included in the analysis. Out of 101 randomized, 78 met this criteria. Of the 78, 3 were excluded for noncompliance, leaving 75 participants. Of these 75, 75 completed the 25%, 73 completed the 50% \& 75 completed the 100% diets.
| Arm | Measure | Value (NUMBER) |
|---|---|---|
| 25% Cho | Difference in Choline Deficiency Signature | 57 percentage participants with signature |
| 50% Cho | Difference in Choline Deficiency Signature | 47 percentage participants with signature |
| 100% Cho | Difference in Choline Deficiency Signature | 23 percentage participants with signature |
p-value: <0.00195% CI: [3.48, 14.7]Fisher Exact
p-value: 0.0195% CI: [2.06, 8.11]Fisher Exact
p-value: 0.08695% CI: [0.86, 3.58]Fisher Exact
Primary
Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet
Metabolomics was conducted on plasma that was collected from individuals at the end of each 2-week diet period. UHPLC High Resolution Mass Spectrometry was used for differential profiling (PMID: 33415121). Supervised Orthogonal Partial Least Squares Discriminant Analysis was used to determine signals that differentiated the 25% choline dietary group from the 100% choline dietary group. Metabolites that differentiated the 25% and 100% choline dietary groups with variable importance to projection (VIP) \>1 and p-value \< 0.05 are reported. The signals for these metabolites were matched by retention time, exact mass, and MS/MS to standards run on the same platform. Because this is a differential profiling method (not quantitative), the mean and standard deviation of peak intensities detected on the untargeted platform are reported. Results are reported for the selected metabolites for the 25%, 50%, and 100% choline dietary groups. Ratios can be obtained by division of the intensity data.
Time frame: At the end of 2 weeks of respective Cho diet
Population: Participants who completed at least 2 arms were included in the analysis. Out of 101 randomized, 78 met this criteria. Of the 78, 3 were excluded for noncompliance, leaving 75 participants. Of these 75, 75 completed the 25%, 73 completed the 50% \& 75 completed the 100% diets.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| 25% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Gamma-Muricholic Acid | 4285.05 Ion abundance (intensity) | Standard Deviation 5853.57 |
| 25% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | N6-Acetyl-L-lysine | 24100.04 Ion abundance (intensity) | Standard Deviation 7506.44 |
| 25% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Deoxycholic Acid | 731438.14 Ion abundance (intensity) | Standard Deviation 633205.4 |
| 25% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Kynurenate | 3200.20 Ion abundance (intensity) | Standard Deviation 1562.4 |
| 25% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Propionylcarnitine | 104096.43 Ion abundance (intensity) | Standard Deviation 39621.16 |
| 25% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | N-Acetylglycine | 32411.20 Ion abundance (intensity) | Standard Deviation 33107.9 |
| 25% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | L-Carnitine | 1228784.33 Ion abundance (intensity) | Standard Deviation 306909 |
| 25% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Methylimidazoleacetic Acid | 9875.65 Ion abundance (intensity) | Standard Deviation 6304.32 |
| 25% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Valerylcarnitine | 94263.17 Ion abundance (intensity) | Standard Deviation 36626.32 |
| 25% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | DL-2-Aminoadipic Acid | 3088.49 Ion abundance (intensity) | Standard Deviation 1065.78 |
| 25% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | L-Ornithine | 2696.01 Ion abundance (intensity) | Standard Deviation 1213.2 |
| 50% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | DL-2-Aminoadipic Acid | 3065.37 Ion abundance (intensity) | Standard Deviation 1168.13 |
| 50% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | L-Carnitine | 1178435.52 Ion abundance (intensity) | Standard Deviation 316249.45 |
| 50% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Propionylcarnitine | 99820.22 Ion abundance (intensity) | Standard Deviation 48710.67 |
| 50% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Valerylcarnitine | 93531.64 Ion abundance (intensity) | Standard Deviation 44570.42 |
| 50% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Gamma-Muricholic Acid | 4927.62 Ion abundance (intensity) | Standard Deviation 6834.32 |
| 50% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Kynurenate | 3186.66 Ion abundance (intensity) | Standard Deviation 1405.28 |
| 50% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | N-Acetylglycine | 28421.55 Ion abundance (intensity) | Standard Deviation 27012.58 |
| 50% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Deoxycholic Acid | 802858.85 Ion abundance (intensity) | Standard Deviation 772625.96 |
| 50% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | L-Ornithine | 2684.53 Ion abundance (intensity) | Standard Deviation 1338.5 |
| 50% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Methylimidazoleacetic Acid | 10075.93 Ion abundance (intensity) | Standard Deviation 5111.3 |
| 50% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | N6-Acetyl-L-lysine | 24361.81 Ion abundance (intensity) | Standard Deviation 7273.97 |
| 100% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Deoxycholic Acid | 600931.15 Ion abundance (intensity) | Standard Deviation 509303.07 |
| 100% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Valerylcarnitine | 82582.08 Ion abundance (intensity) | Standard Deviation 40040.76 |
| 100% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | DL-2-Aminoadipic Acid | 2805.00 Ion abundance (intensity) | Standard Deviation 981.61 |
| 100% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | L-Ornithine | 2410.86 Ion abundance (intensity) | Standard Deviation 1223.7 |
| 100% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Propionylcarnitine | 80011.86 Ion abundance (intensity) | Standard Deviation 38878.77 |
| 100% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | N6-Acetyl-L-lysine | 22522.45 Ion abundance (intensity) | Standard Deviation 5552.79 |
| 100% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Kynurenate | 2739.41 Ion abundance (intensity) | Standard Deviation 1362.3 |
| 100% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Methylimidazoleacetic Acid | 8510.57 Ion abundance (intensity) | Standard Deviation 4212.85 |
| 100% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | N-Acetylglycine | 26644.26 Ion abundance (intensity) | Standard Deviation 26448.88 |
| 100% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | Gamma-Muricholic Acid | 2519.32 Ion abundance (intensity) | Standard Deviation 4817.63 |
| 100% Cho | Ion Abundance (Intensity) of Metabolites as Indicators of the Intake of 25%, 50%, or 100% Choline in the Diet. The Ratio of the Intensity of Metabolite Signals for Each Dietary Group Can be Calculated and Correlated With the Level of Choline in the Diet | L-Carnitine | 1012630.35 Ion abundance (intensity) | Standard Deviation 344564.26 |
p-value: <0.049Paired 2-sided t-test
Primary
Ratio of Liver Choline Pool Size by Isotope Dilution
The liver choline pool determined by the dilution of the deuterated choline metabolites formed in liver and released to plasma as measured by isotopic enrichment ratio (IER). The IER for a given metabolite is defined as the concentration of a deuterated metabolite divided by the sum of deuterated and non-deuterated metabolite.
Time frame: 24 hours following administration of choline-d9 on day 12 of respective dietary intervention
Population: Participants who completed at least 2 arms were included in the analysis. Out of 101 randomized, 78 met this criteria. Of the 78, 3 were excluded for noncompliance, leaving 75 participants. Of these 75, 75 completed the 25%, 73 completed the 50% \& 75 completed the 100% diets. One participant completing 3 arms was missing 50% isotope dilution data; consequently, outcomes analyzed for 75 participants (25%), 72 participants (50%), \& 75 participants (100%).
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| 25% Cho | Ratio of Liver Choline Pool Size by Isotope Dilution | Choline IER | 0.00023 isotopic enrichment ratio | Standard Deviation 0.00511 |
| 25% Cho | Ratio of Liver Choline Pool Size by Isotope Dilution | Betaine IER | 0.02357 isotopic enrichment ratio | Standard Deviation 0.00802 |
| 25% Cho | Ratio of Liver Choline Pool Size by Isotope Dilution | Phosphatidylcholine IER | 0.01158 isotopic enrichment ratio | Standard Deviation 0.00326 |
| 50% Cho | Ratio of Liver Choline Pool Size by Isotope Dilution | Choline IER | 0.00132 isotopic enrichment ratio | Standard Deviation 0.00493 |
| 50% Cho | Ratio of Liver Choline Pool Size by Isotope Dilution | Betaine IER | 0.02403 isotopic enrichment ratio | Standard Deviation 0.00882 |
| 50% Cho | Ratio of Liver Choline Pool Size by Isotope Dilution | Phosphatidylcholine IER | 0.01130 isotopic enrichment ratio | Standard Deviation 0.00324 |
| 100% Cho | Ratio of Liver Choline Pool Size by Isotope Dilution | Betaine IER | 0.02184 isotopic enrichment ratio | Standard Deviation 0.00574 |
| 100% Cho | Ratio of Liver Choline Pool Size by Isotope Dilution | Phosphatidylcholine IER | 0.01017 isotopic enrichment ratio | Standard Deviation 0.00234 |
| 100% Cho | Ratio of Liver Choline Pool Size by Isotope Dilution | Choline IER | 0.00191 isotopic enrichment ratio | Standard Deviation 0.00399 |
Comparison: Betaine IERp-value: 0.0247Repeated measures ANOVA
Comparison: Choline IERp-value: <0.0001Repeated measures ANOVA
Comparison: Phosphatidylcholine IERp-value: 0.0002Repeated measures ANOVA
Secondary
Validation of Isotope Dilution Method to Assess Choline Pool Size by Magnetic Resonance Spectroscopy (MRS)
MRS is a direct measurement of liver choline content. Changes in liver choline by MRS should correlate with changes in liver choline inferred by calculation of isotope enrichment ratio (IER) of plasma metabolites. Pearson correlation coefficient used to study the correlation between data generated from the two types of measurements.
Time frame: At the end of 2 weeks of respective Cho diet
Population: MRS was run on only an initial 40 participants due to limited funding. Due to extensive variation, MRS data were not usable for 8 participants in the 25%, 10 participants in the 50%, and 8 participants in the 100% Cho arms.
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| 25% Cho | Validation of Isotope Dilution Method to Assess Choline Pool Size by Magnetic Resonance Spectroscopy (MRS) | 8.4 mM | Standard Deviation 4.1 |
| 50% Cho | Validation of Isotope Dilution Method to Assess Choline Pool Size by Magnetic Resonance Spectroscopy (MRS) | 7.8 mM | Standard Deviation 4.5 |
| 100% Cho | Validation of Isotope Dilution Method to Assess Choline Pool Size by Magnetic Resonance Spectroscopy (MRS) | 11.2 mM | Standard Deviation 7.2 |
Comparison: Liver choline concentration vs. betaine IERp-value: 0.2351Pearson's Correlation Coefficient
Comparison: Liver choline concentration vs. Phosphatidylcholine IERp-value: 0.4501Pearson's Correlation Coefficient
Comparison: Liver choline concentration vs. Choline IERp-value: 0.4136Pearson's Correlation Coefficient
Comparison: Liver choline concentration vs. Betaine IERp-value: 0.9463Pearson's Correlation Coefficient
Comparison: Liver choline concentration vs. Phosphatidylcholine IERp-value: 0.0675Pearson's Correlation Coefficient
Comparison: Liver choline concentration vs. Choline IERp-value: 0.2863Pearson's Correlation Coefficient
Comparison: Liver choline concentration vs. Betaine IERp-value: 0.5552Pearson's Correlation Coefficient
Comparison: Liver choline concentration vs. Phosphatidylcholine IERp-value: 0.4209Pearson's Correlation Coefficient
Comparison: Liver choline concentration vs. Choline IERp-value: 0.6647Pearson's Correlation Coefficient
Other Pre-specified
SNPs That Create Inefficiencies in Choline Metabolism Associated With Change in Choline Pool Size and Choline Biomarkers
Exploratory analysis of \>2 million SNPs measured on a custom Illumina Expanded Multi-Ethnic genotyping array (Mega-Ex). The same analysis described for Outcome 4 will be applied for Outcome 7. Benjamini-Hochberg method for False Discovery Rate (FDR) correction will be used for multiple testing correction.
Time frame: At baseline visit