Plasma Betaine Research Articles

Whole egg consumption increases plasma choline and betaine without affecting TMAO levels and gut microbiome in overweight postmenopausal woman

As a crucial part of the symbiotic system, the gut microbiome has been shown to be a metabolic organ that presents important connections to many diseases and conditions, including cardiovascular diseases (CVD), the world wide leading cause of death. Trimethylamine N‐oxide (TMAO) is a plasma metabolite that is positively correlated with CVD. Trimethylamine (TMA) is produced by gut bacteria from dietary choline, betaine, or L‐carnitine, and is then converted in the liver to TMAO, which in turn affects hepatic and intestinal lipid and cholesterol metabolism. Because eggs are rich in choline, it has been speculated that their consumption may increase plasma TMAO. The objective of this study was to examine the effect of two eggs per day on plasma TMAO level and how it is related with gut microbiome composition in mildly hypercholesterolemic postmenopausal women. In this randomized, cross‐over study, 20 overweight human subjects were given two whole eggs and the equivalent amount of yolk‐free substitute as breakfast for four weeks, in randomized order, with a four‐week washout in between. Fasting blood draws and stool were collected at the beginning and of each treatment period. Plasma TMAO, choline, betaine and other metabolites were analyzed using LC/MS, while gut microbiome composition was analyzed using 16S amplicon sequencing. Plasma choline and betaine were significantly increased after whole egg but not yolk‐free substitute, however TMAO level was not significantly affected by treatments. Gut microbiome composition showed large inter‐individual variability at baseline and in response to the treatments. Parabacteroides, Ruminococcus, and Faecalibacterium were slightly increased after whole egg but not yolk‐free substitute for the majority of the subjects, while Bilophila slightly increased after yolk‐free substitute but not whole egg. The consumption of two eggs per day in overweight, postmenopausal mildly hypercholesterolemic women significantly increased plasma choline and betaine, but not TMAO, with a subtle effect on gut microbiome composition.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Betaine insufficiency and reduced betaine dependent remethylation are associated with hyperhomocysteinemia, oxidative stress and mitochondrial damage in early Polycystic Kidney Disease

Polycystic kidney disease (PKD) is the most common monogenic kidney disorder and an important cause of end stage renal disease. Vascular abnormalities are the most important non‐cystic complication and contribute to renal disease progression. Homocysteine (Hcy)‐mediated endothelial dysfunction (ED) and oxidative stress are evident early on, but the underlying mechanisms remain unknown. We hypothesized that abnormalities in Hcy metabolism are responsible for the increased levels, leading to Hcy‐induced ED from early stages of PKD.MethodsWe investigated the Hcy metabolic pathway (transulfuration/remethylation fig. 1) and downstream effects ‐main sources of renal tubular (RT) and endothelial cell (EC) reactive oxygen species (ROS), NADPH oxidase 4, (NOX4) and mitochondria‐ in 4‐week‐old PCK and Sprague‐Dawley (SD) control rats (n=12 each). Kidneys were harvested, frozen in liquid nitrogen, or preserved in formalin for metabolomics and ex‐vivo studies. Twenty‐four‐hour urine and terminal blood samples were collected for metabolite analysis and chemistries. Plasma folic acid, vitamin B12, and Hcy were determined by ELISA. Hcy pathway metabolites (betaine, dimethylglycine) were determined by 1HNMR. Endothelial NO (eNOS) and oxidative stress were assessed by double immunofluorescence staining for CD31/eNOS and CD31/NOX4, respectively. RT and EC mitochondrial structure was assessed by electron microscopy.ResultsSerum creatinine and BUN remained unchanged, yet kidney weight/body weight ratio were increased in PCK. eNOS immunoreactivity was lower in PCK vs SD. Plasma Hcy and urine excretion of methyl donor betaine were elevated in PCK, and Hcy positively correlated with urine betaine. Plasma betaine was similar between the groups, but its tissue concentration was lower in PCK, which may lead to betaine insufficiency, impaired betaine dependent remethylation, and hyperhomocysteinemia. Contrarily, tissue glutathione concentration was higher in PCK vs SD, and plasma folate and vitamin B12 were similar, arguing against a defect in the transsulfuration or folate‐dependent remethylation pathways. RT NOX4 expression and immunoreactivity were increased in PCK compared to SD, in association with renal oxidative stress. PCK had increased number of CD31/NOX4 positive cells in peritubular capillaries, indicating increased EC ROS production. Collecting duct and EC mitochondria were swollen and showed cristae remodeling.ConclusionIn PKD, early elevation in Hcy is likely related to betaine insufficiency and impaired betaine‐dependent remethylation may be responsible for the early elevation of Hcy. NOX4 and mitochondria abnormalities may be important contributors to oxidative stress and disease severity. Our data suggests these may not only be early vascular disease markers, but also play an important pathogenic role in vascular complications associated with PKD, and contribute to renal disease progression.Support or Funding InformationR25‐DK101405, NIDDK P30 DK090728, NIDDK P30 DK 106912This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Maternal betaine status, but not that of choline or methionine, is inversely associated with infant birth weight.

Maternal one-carbon metabolism during pregnancy is crucial for fetal development and programming by DNA methylation. However, evidence on one-carbon biomarkers other than folate is lacking. We, therefore, investigated whether maternal plasma methyl donors, that is, choline, betaine and methionine, are associated with birth outcomes. Blood samples were obtained from 115 women during gestation (median 26·3 weeks, 90 % range 22·7-33·0 weeks). Plasma choline, betaine, methionine and dimethylglycine were measured using HPLC-tandem MS. Multivariate linear and logistic regression models were used to estimate the association between plasma biomarkers and birth weight, birth length, the risk of small-for-gestational-age and large-for-gestational-age (LGA). Higher level of maternal betaine was associated with lower birth weight (-130·3 (95 % CI -244·8, -15·9) per 1 sd increment for log-transformed betaine). Higher maternal methionine was associated with lower risk of LGA, and adjusted OR, with 95 % CI for 1 sd increase in methionine concentration was 0·44 (95 % CI 0·21, 0·89). Stratified analyses according to infant sex or maternal plasma homocysteine status showed that reduction in birth weight in relation to maternal betaine was only limited to male infants or to who had higher maternal homocysteine status (≥5·1 µmol/l). Higher maternal betaine status was associated with reduced birth weight. Maternal methionine was inversely associated with LGA risk. These findings are needed to be replicated in future larger studies.

Plasma choline, homocysteine and vitamin status in healthy adults supplemented with krill oil: a pilot study

Plasma concentrations of metabolites along the choline oxidation and tryptophan degradation pathways have been linked to lifestyle diseases and dietary habits. This study aimed to investigate how krill oil, a source of ω-3 polyunsaturated fatty acids (PUFAs) with a high phosphatidylcholine content, affected these parameters. The pilot study was conducted as a 28 days intervention in 17 healthy volunteers (18–36 years), who received a supplement of 4.5 g krill oil per day, providing 833 mg ω-3 PUFAs, and 1750 mg phosphatidylcholine. Krill oil supplementation increased fasting plasma choline (+28.4%, p < .001), betaine (+26.6%, p < .001), dimethylglycine (+33.7%, p < .001) and sarcosine (+16.8%, p < .001), whereas no statistically significant changes were seen for plasma glycine, serine, methionine, total homocysteine, cysteine, cystathionine, methionine sulfoxide, folate, cobalamin, B2-, B3-, and B6 vitamers, tryptophan, kynurenines, nicotinamide, vitamin A and vitamin E. In summary, krill oil supplementation influenced choline metabolite levels, but not plasma metabolites of the tryptophan-kynurenine-nicotinamide pathways and vitamins. These observations should be confirmed in a placebo-controlled trial, including an ω-3 PUFA supplement without phospholipids to explore the potential additive effects of the different active ingredients.

The Role of Plasma Choline and Betaine in Cardiovascular Disease Risk

Background: As precursors of Trimethylamine N-oxide (TMAO), the association between dietary choline and betaine and the risk of CVD have been well studied. However, researches on the relationship between plasma choline and betaine and the development and progression of CVD remain inconclusive. Methods: PubMed was used to search for the studies on association between choline and betaine in plasma and CVD risk. A systemic meta-analysis was then conducted. Importantly, a case-control study including 2,019 subjects was further performed to validate the relationship between plasma choline and betaine and the progression of CVD. Findings: A total of four studies enrolled 7,219 subjects were eventually included in the meta-analysis. The result suggested plasma choline was significantly associated with CVD progression [Hazard ratio (HR): 2·33, 95% Confidence interval (CI): 1·32-4·12, p = 3·50E-3], while plasma betaine not (HR: 1·14, 95% CI: 0·89-1·45, p = 2·91E-1). Furthermore, 2,019 subjects including 235 normal coronary artery (NCA), 631 non-significant coronary artery disease (nos-CVD) and 1,153 acute coronary syndrome (ACS) individuals, were recruited to confirm the findings. The result adjusted multiple covariates and TMAO indicated that plasma choline was significantly associated with CVD progression [NCA vs. nos-CVD, Odds ratio (OR): 1·75, 95% CI: 1·08 - 2·83, p = 2·24E-02]. Interpretation: Increased plasma choline level associates with the risk of CVD. The plasma choline contributes to CVD progression from NCA to nos-CVD independent from TMAO. However, plasma choline contributes to CVD progression by converting into TMAO from nos-CVD to ACS. Funding: This study was supported by National Natural Science Foundation of China (91639115). Declaration of Interest: Authors declare no conflicts of interest. Ethical Approval: Participants provided written informed consent and the China Pharmaceutical University and Jiangsu University Review Board approved the study.

Variations in plasma choline and metabolite concentrations in healthy adults

BackgroundPlasma concentrations of choline and its metabolites might serve as biomarkers for the health outcomes of several pathological states such as cardiovascular disease and cancer. However, information about the reliability of biomarkers of choline status is limited. We investigated biological variations in repeated measures of choline and metabolites in healthy adults to assess them as biomarkers. MethodsBlood samples were collected after an overnight fast at three-time points 12 days apart from 40 adults (mean age, 33 y; male, n = 21). A subset (n = 19; [male, n = 8]) provided one additional sample after a breakfast meal. Plasma free choline, betaine and dimethylglycine were measured using liquid chromatography-tandem mass spectrometry, and plasma phosphatidylcholine, sphingomyelin and lysophosphatidylcholine were measured using high-performance liquid chromatography. ResultsThe biological variations observed for choline and metabolites were ≤ 13% for adult fasting samples. This corresponded to intra-class correlations (ICC) that ranged from 0.593 to 0.770 for fasting values for choline and metabolites. A similar ICC range was also obtained between fasting and post-prandial states. Although most post-prandial concentrations of choline and metabolites were significantly higher (P < .05) than fasting, all fell within a calculated reference interval. The participants were correctly classified in tertiles for fasting and post-prandial states for choline (68%) and metabolites (range = 32% phosphatidylcholine and 79% for sphingomyelin). ConclusionsThese findings indicate that biological variations of choline and metabolites are low in healthy adults and values from a single blood sample can be used as a biomarker. However, choosing phosphatidylcholine as a biomarker is less reliable.

Plasma Betaine Is Positively Associated with Developmental Outcomes in Healthy Toddlers at Age 2 Years Who Are Not Meeting the Recommended Adequate Intake for Dietary Choline

Choline is an important nutrient during development. However, there are limited data on dietary choline intake and status in toddlers and the relation to neurodevelopmental outcomes. This study assessed dietary choline intake and status in healthy toddlers at ages 1 and 2 y and determined the relation to neurodevelopmental outcomes. This is a secondary analysis of data from healthy toddlers enrolled in a double-blind, randomized controlled trial of long-chain polyunsaturated fatty acid supplementation between ages 1 and 2 y. Dietary intakes of betaine and choline were estimated by 3-d food records; plasma free choline, betaine, and dimethylglycine were quantified by liquid chromatography-tandem mass spectrometry. Developmental outcomes were assessed at age 2 y with the use of the Bayley Scales of Infant and Toddler Development, 3rd edition (Bayley-III), Cognitive and Language composites, and the Beery-Buktenica Developmental Test of Visual-Motor Integration (Beery-VMI). The mean±SD daily intake for total choline at age 1 y was 174±56.2 mg/d and increased (P<0.001) to 205±67.5 mg/d at age 2 y. At ages 1 and 2 y, 71.8% and 55.8%, respectively, of toddlers did not meet the recommended 200-mg/d Adequate Intake (AI) for dietary choline. At age 1 y, mean±SD plasma free choline, betaine, and dimethylglycine concentrations were 10.4±3.3, 41.1±15.4, and 4.1±1.9 µmol/L, respectively. Plasma free choline (8.5±2.3 µmol/L) and dimethylglycine (3.2±1.3 µmol/L) concentrations were lower (P<0.001) at age 2 y. Plasma betaine concentrations were positively associated with the Beery-VMI (β=0.270; 95% CI: 0.026, 0.513; P=0.03) at age 2 y. These findings suggest that most toddlers are not meeting the recommended AI for dietary choline and that higher plasma betaine concentrations are associated with better visual-motor development at age 2 y. Further work is required to investigate choline metabolism and its role in neurodevelopment in toddlers. The trial is registered at clinicaltrials.gov as NCT01263912.

Abstract 3268: Associations of plasma trimethylamine N-oxide (TMAO), choline, and betaine with the gut microbiome and biomarkers of inflammation and cardiometabolic risk in the Multiethnic Cohort

Abstract Background: TMAO is produced by gut microbial metabolism of dietary quaternary amines and has been linked to several chronic diseases, including colorectal cancer and cardiovascular disease. However, the microbes and mechanisms involved are still unclear. Here, we aimed to 1) identify microbial taxa associated with TMAO and 2) evaluate associations of plasma TMAO, along with its precursors choline and betaine, with inflammatory and cardiometabolic biomarkers. Methods: We conducted a cross-sectional analysis in 1,716 participants (men and women, 60-77, and of 5 racial/ethnic groups) of the Adiposity Phenotype Study within the Multiethnic Cohort in Hawaii and California. Plasma TMAO, choline, and betaine were measured by LC-MS/MS. The gut microbiome was analyzed by Illumina MiSeq paired-end sequencing of the 16S rRNA gene V1-V3 region using DNA extracted from stool. Fasting blood was analyzed for lipopolysaccharide-binding protein (LBP) and insulin by ELISA; C-reactive protein (CRP), cholesterol, glucose, and triglycerides using a Cobas autoanalyzer; and HOMA-IR for insulin resistance by derivation. Associations between microbial genera and TMAO were assessed with negative binomial and zero-inflated negative binomial (ZINB) regression models, with control of the false discovery rate (FDR) using the Benjamini-Hochberg procedure. Associations of TMAO, choline, and betaine with biomarkers were determined using multivariable linear regression and a ZINB model for CRP, adjusted for age, sex, race/ethnicity, physical activity, aspirin use, and % body fat. Results: Of 105 genera examined, 7 were associated with TMAO at an FDR of 0.01: Streptoccoccus, Blautia, Clostridium, a genus from the Ruminococcaceae family, Butyricimonas, Lactobacillus, and an uncultured Verrucomicrobia of the order vadinHA64; all showed a positive relationship. Plasma choline (β±SE: 0.23±0.10; p=0.03) and betaine (0.19±0.08; p=0.02), but not TMAO, were positively associated with CRP, whereas only choline was associated with LBP (1.97±0.80; p=0.01). Choline was inversely associated with HDL cholesterol (-14.86±2.29; p&amp;lt;0.001) and positively associated with LDL cholesterol (9.86±5.01; p=0.049). Betaine (-0.16±0.03; p&amp;lt;0.001) and choline (0.14±0.05; p=0.002) were associated with triglycerides, while TMAO was not (0.01±0.02; p=0.46). All 3 plasma compounds were associated with HOMA-IR, with only betaine having an inverse association (-0.32±0.06; p=0.004). Conclusion: We found several genera of gut microbes associated with plasma TMAO, including Clostridium, several species of which can metabolize choline to trimethylamine. We also identified associations between plasma TMAO, choline, and betaine with inflammatory and cardiometabolic markers, particularly choline, which indicated risk to adverse health outcomes. Citation Format: Benjamin C. Fu, Meredith AJ Hullar, Timothy W. Randolph, Adrian A. Franke, Kristine R. Monroe, Iona Cheng, Lynne R. Wilkens, John Shepherd, Loïc Le Marchand, Unhee Lim, Johanna W. Lampe. Associations of plasma trimethylamine N-oxide (TMAO), choline, and betaine with the gut microbiome and biomarkers of inflammation and cardiometabolic risk in the Multiethnic Cohort [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3268.

Plasma trimethylamine-N-oxide and related metabolites are associated with type 2 diabetes risk in the Prevención con Dieta Mediterránea (PREDIMED) trial

The role of trimethylamine-N-oxide (TMAO) in type 2 diabetes (T2D) is currently partially understood and controversial. The aim of this study was to investigate associations between TMAO and related metabolites with T2D risk in subjects at high risk of cardiovascular disease. This is a case-cohort design study within the Prevención con Dieta Mediterránea (PREDIMED) study, with 251 incident T2D cases and a random sample of 694 participants (641 noncases and 53 overlapping cases) without T2D at baseline (median follow-up: 3.8 y). We used liquid chromatography-tandem mass spectrometry to measure plasma TMAO, l-carnitine, betaine, lyso-phosphatidylcholine (LPC) and lyso-phosphatidylethanolamine (LPE) species, phosphocholine, α-glycerophosphocholine, and choline at baseline and after 1 y. We examined associations with the use of weighted Cox proportional hazard models, accounting for the weighted case-cohort design by the Barlow method. After adjustment for recognized T2D risk factors and multiple testing, individuals in the highest quartile of baseline TMAO and α-glycerophosphocholine had a lower risk of T2D [HR (95% CI): 0.52 (0.29, 0.89) and 0.46 (0.24, 0.89), respectively]. The HR (95% CI) comparing the extreme quartiles of betaine was 0.41 (0.23, 0.74). Similar trends were observed for C16:0 LPC, C18:1 LPC, C18:0 LPC, C20:4 LPC, C22:6 LPC, C18:1 LPC plasmalogen, and C16:0 LPE. After correcting for multiple comparisons, participants in the highest quartile of 1-y changes in oleic acid LPC plasmalogen concentrations had a lower T2D risk than the reference quartile. Whether the associations between plasma TMAO and certain metabolite concentrations with T2D risk reflect its pathophysiology or represent an epiphenomenon needs to be elucidated. This trial is registered at http://www.controlled-trials.com as ISRCTN35739639.

Betaine or folate can equally furnish remethylation to methionine and increase transmethylation in methionine-restricted neonates

Methionine partitioning between protein turnover and a considerable pool of transmethylation precursors is a critical process in the neonate. Transmethylation yields homocysteine, which is either oxidized to cysteine (i.e., transsulfuration), or is remethylated to methionine by folate- or betaine- (from choline) mediated remethylation pathways. The present investigation quantifies the individual and synergistic importance of folate and betaine for methionine partitioning in neonates. To minimize whole body remethylation, 4–8-d-old piglets were orally fed an otherwise complete diet without remethylation precursors folate, betaine and choline (i.e. methyl-deplete, MD-) (n=18). Dietary methionine was reduced from 0.3 to 0.2 g/(kg∙d) on day-5 to limit methionine availability, and methionine kinetics were assessed during a gastric infusion of [13C1]methionine and [2H3-methyl]methionine. Methionine kinetics were reevaluated 2 d after pigs were rescued with either dietary folate (38 μg/(kg∙d)) (MD + F) (n=6), betaine (235 mg/(kg∙d)) (MD + B) (n=6) or folate and betaine (MD + FB) (n=6). Plasma choline, betaine, dimethylglycine (DMG), folate and cysteine were all diminished or undetectable after 7 d of methyl restriction (P<.05). Post-rescue, plasma betaine and folate concentrations responded to their provision, and homocysteine and glycine concentrations were lower (P<.05). Post-rescue, remethylation and transmethylation rates were~70–80% higher (P<.05), and protein breakdown was spared by 27% (P<.05). However, rescue did not affect transsulfuration (oxidation), plasma methionine, protein synthesis or protein deposition (P>.05). There were no differences among rescue treatments; thus betaine was as effective as folate at furnishing remethylation. Supplemental betaine or folate can furnish the transmethylation requirement during acute protein restriction in the neonate.

L-5-Methyltetrahydrofolate Supplementation Increases Blood Folate Concentrations to a Greater Extent than Folic Acid Supplementation in Malaysian Women

Folic acid fortification of grains is mandated in many countries to prevent neural tube defects. Concerns regarding excessive intakes of folic acid have been raised. A synthetic analog of the circulating form of folate, l-5-methyltetrahydrofolate (l-5-MTHF), may be a potential alternative. The objective of this study was to determine the effects of folic acid or l-5-MTHF supplementation on blood folate concentrations, methyl nutrient metabolites, and DNA methylation in women living in Malaysia, where there is no mandatory fortification policy. In a 12-wk, randomized, placebo-controlled intervention trial, healthy Malaysian women (n=142, aged 20-45 y) were randomly assigned to receive 1 of the following supplements daily: 1 mg (2.27 μmol) folic acid, 1.13 mg (2.27 μmol) l-5-MTHF, or a placebo. The primary outcomes were plasma and RBC folate and vitamin B-12 concentrations. Secondary outcomes included plasma total homocysteine, total cysteine, methionine, betaine, and choline concentrations and monocyte long interspersed nuclear element-1 (LINE-1) methylation. The folic acid and l-5-MTHF groups had higher (P<0.001) RBC folate (mean±SD: 1498±580 and 1951±496 nmol/L, respectively) and plasma folate [median (25th, 75th percentiles): 40.1 nmol/L (24.9, 52.7 nmol/L) and 52.0 nmol/L (42.7, 73.1 nmol/L), respectively] concentrations compared with RBC folate (958±345 nmol/L) and plasma folate [12.6 nmol/L (8.80, 17.0 nmol/L)] concentrations in the placebo group at 12 wk. The l-5-MTHF group had higher RBC folate (1951±496 nmol/L; P=0.003) and plasma folate [52.0 nmol/L (42.7, 73.1 nmol/L); P=0.023] at 12 wk than did the folic acid group [RBC folate, 1498±580 nmol/L; plasma folate, 40.1 nmol/L (24.9, 52.7 nmol/L)]. The folic acid and l-5-MTHF groups had 17% and 15%, respectively, lower (P<0.001) plasma total homocysteine concentrations than did the placebo group at 12 wk; there were no differences between the folic acid and l-5-MTHF groups. No differences in plasma vitamin B-12, total cysteine, methionine, betaine, and choline and monocyte LINE-1 methylation were observed. These findings suggest differential effects of l-5-MTHF compared with folic acid supplementation on blood folate concentrations but no differences on plasma total homocysteine lowering in Malaysian women. This trial was registered at clinicaltrials.gov as NCT01584050.

Fetal one-carbon nutrient concentrations may be affected by gestational diabetes

Both insufficiency and excess of one-carbon nutrients (folate, choline, vitamins B6 and B12) during pregnancy have been associated with gestational diabetes mellitus (GDM). However, the precise nature of this association has not been clearly established. We hypothesized that GDM may affect one-carbon nutrients concentrations in the fetus, thus possibly participating in epigenetic programing of the offspring. Maternal blood was collected at recruitment (12–16 weeks). At delivery (28–42 weeks), both maternal and cord blood were collected. Blood concentrations of one-carbon nutrients and their metabolites were compared between the two groups. A total of 368 women were included in the study, of whom 19 (5.6%) were later diagnosed with GDM. No significant differences were found in maternal blood concentrations of one-carbon nutrients and their metabolites between the GDM and control groups at recruitment or at delivery. In cord blood, however, serum folate (87.7 [IQR 70.4–103.9] vs 66.6 [IQR 45.5–80.3] nmol/L, P = .025) and plasma TMAO (2.82 [IQR 1.3–3.2] vs 1.35 [IQR 1.0–2.0] μmol/L, P = .017) concentrations were higher, while plasma betaine concentrations were lower (17.5 [IQR 16.3–19.4] vs 21.1 [IQR 18.0–24.1] μmol/L, P = .019) in infants born to mothers with GDM compared with control. Our data suggest that while maternal blood concentrations of one-carbon nutrients and their metabolites may not affect the risk of GDM, GDM may alter concentrations of serum folate, plasma betaine and TMAO in cord blood. These alterations in one-carbon nutrient concentrations in fetal circulation may impact epigenetic programing, thereby contributing to physiologic changes and disease susceptibility in adulthood associated with GDM offspring.

Effects of Probiotic Supplementation on Trimethylamine-N-Oxide Plasma Levels in Hemodialysis Patients: a Pilot Study.

Components present in the diet, L-carnitine, choline, and betaine are metabolized by gut microbiota to produce metabolites such as trimethylamine-N-oxide (TMAO) that appear to promote cardiovascular disease in chronic kidney disease (CKD) patients. The objective of this pilot study was to evaluate the effects of probiotic supplementation for 3months on plasma TMAO levels in CKD patients on hemodialysis (HD). A randomized, double-blind trial was performed in 21 patients [54.8 ± 10.4years, nine men, BMI 26.1 ± 4.8kg/m2, dialysis vintage 68.5 (34.2-120.7) months]. Ten patients were randomly allocated to the placebo group and 11 to the probiotic group [three capsules, totaling 9 × 1013 colony-forming units per day of Streptococcus thermophilus (KB19), Lactobacillus acidophilus (KB27), and Bifidobacteria longum (KB31). Plasma TMAO, choline, and betaine levels were measured by LC-MS/MS at baseline and after 3months. While TMAO did not change after probiotic supplementation, there was a significant increase in betaine plasma levels. In contrast, the placebo group showed a significant decrease in plasma choline levels. Short-term probiotic supplementation does not appear to influence plasma TMAO levels in HD patients. Long-term studies are needed to determine whether probiotics may affect TMAO production in CKD patients.

Association of betaine with blood pressure in dialysis patients.

Mechanisms underlying elevated blood pressure in dialysis patients are complex as a variety of non-traditional factors are involved. We sought to explore the association of circulating betaine, a compound widely distributed in food, with blood pressure in dialysis patients. We used baseline data of an ongoing cohort study involving patients on hemodialysis. Plasma betaine was measured by high performance liquid chromatography in 327 subjects. Blood pressure level was determined by intradialytic ambulatory blood pressure monitoring. The mean age of the patients was 52.6±11.9years, and 58.4% were male. Average interdialytic ambulatory systolic and diastolic blood pressure were 138.4±22.7mm Hg and 84.4±12.5mm Hg, respectively. Mean plasma betaine level was 37.6μmol/L. Multiple linear regression analysis revealed significant associations of betaine with both systolic blood pressure (β=-3.66, P=.003) and diastolic blood pressure (β=-2.00, P=.004). The associations persisted even after extensive adjustment for cardiovascular covariates. Subgroup analysis revealed that the association between betaine and blood pressure was mainly limited to female patients. Our data suggest that alteration of circulating betaine possibly contributes to blood pressure regulation in these patients.

Choline Protects Against Intestinal Failure-Associated Liver Disease in Parenteral Nutrition-Fed Immature Rats.

Deficiency of choline, a required nutrient, is related to intestinal failure-associated liver disease (IFALD). Therefore, we aimed to investigate the effects of choline supplementation on IFALD and the underlying mechanisms. Male Sprague-Dawley rats (4 weeks old) were fed AIN-93G chow and administered intravenous 0.9% saline (control), parenteral nutrition (PN), or PN plus intravenous choline (600 mg/kg) for 7 days. We evaluated body weight, hepatic histology, biochemical indicators, triglycerides, oxidative status, methylation levels of peroxisomal proliferator-activated receptor alpha (PPARα) gene promoter, expression of PPARα and carnitine palmitoyltransferase 1 (CPT1), and levels of choline metabolites. The PN + choline group exhibited improved body weight compared with the PN group. PN impaired hepatic function, increased hepatic triglycerides, induced dyslipidemia, enhanced reactive oxygen species and malondialdehyde, and reduced total antioxidant capacity. The PN group had higher pathologic scores than the control group. These results were prevented by choline administration. Compared with the control group, PN increased PPARα promoter methylation and hepatic betaine concentration, reduced hepatic choline and phosphatidylcholine (PC) levels, decreased plasma choline and betaine concentrations, and downregulated PPARα and CPT1 mRNA and protein expression. Choline supplementation elevated hepatic choline and PC levels and enhanced plasma choline, betaine, and PC concentrations but reduced hepatic betaine level, reversed PPARα promoter hypermethylation, and upregulated PPARα and CPT1 mRNA and protein expression in PN-fed rats, compared with rats receiving PN alone. Choline addition to PN may prevent IFALD by reducing oxidative stress, enhancing hepatic fat export, and promoting fatty acid catabolism in immature rats receiving PN.

Choline and its metabolites are differently associated with cardiometabolic risk factors, history of cardiovascular disease, and MRI-documented cerebrovascular disease in older adults ,

Background: There is a potential role of choline in cardiovascular and cerebrovascular disease through its involvement in lipid and one-carbon metabolism.Objective: We evaluated the associations of plasma choline and choline-related compounds with cardiometabolic risk factors, history of cardiovascular disease, and cerebrovascular pathology.Design: A cross-sectional subset of the Nutrition, Aging, and Memory in Elders cohort who had undergone MRI of the brain (n = 296; mean ± SD age: 73 ± 8.1 y) was assessed. Plasma concentrations of free choline, betaine, and phosphatidylcholine were measured with the use of liquid-chromatography-stable-isotope dilution-multiple-reaction monitoring-mass spectrometry. A volumetric analysis of MRI was used to determine the cerebrovascular pathology (white-matter hyperintensities and small- and large-vessel infarcts). Multiple linear and logistic regression models were used to examine relations of plasma measures with cardiometabolic risk factors, history of cardiovascular disease, and radiologic evidence of cerebrovascular pathology.Results: Higher concentrations of plasma choline were associated with an unfavorable cardiometabolic risk-factor profile [lower high-density lipoprotein (HDL) cholesterol, higher total homocysteine, and higher body mass index (BMI)] and greater odds of large-vessel cerebral vascular disease or history of cardiovascular disease but lower odds of small-vessel cerebral vascular disease. Conversely, higher concentrations of plasma betaine were associated with a favorable cardiometabolic risk-factor profile [lower low-density lipoprotein (LDL) cholesterol and triglycerides] and lower odds of diabetes. Higher concentrations of plasma phosphatidylcholine were associated with characteristics of both a favorable cardiometabolic risk-factor profile (higher HDL cholesterol, lower BMI, lower C-reactive protein, lower waist circumference, and lower odds of hypertension and diabetes) and an unfavorable profile (higher LDL cholesterol and triglycerides).Conclusion: Choline and its metabolites have differential associations with cardiometabolic risk factors and subtypes of vascular disease, thereby suggesting differing roles in the pathogenesis of cardiovascular and cerebral large-vessel disease compared with that of small-vessel disease.

Maternal choline status during pregnancy, but not that of betaine, is related to antenatal mental well-being: The growing up in Singapore toward healthy outcomes cohort.

Choline and betaine status have previously been associated with symptoms of depression. However, the relation of maternal plasma choline and betaine concentrations in pregnancy to peripartum maternal mood is unknown. Maternal plasma choline and betaine concentrations (μmol/L) were measured at 26-28 weeks gestation in the Growing Up in Singapore Toward healthy Outcomes (GUSTO) mother-offspring cohort. Participants completed the State-Trait Anxiety Inventory (STAI) and Edinburgh Postnatal Depression Scale (EDPS) at 26-28 weeks gestation (n = 949) and at 3 months postnatal (n = 689): higher scores are indicative of more symptoms of anxiety and depression. Multivariate linear regression models were used to estimate the association of choline and betaine with ante- and postnatal mental well-being adjusting for covariates. Mean (SD) antenatal plasma choline and betaine concentrations were 9.2 μmol/L (1.6) and 13.1 μmol/L (2.7), respectively. Plasma choline concentrations were positively associated with antenatal depressive (β = .24 EPDS score [95% CI: 0.05-0.43] per μmol/L] and anxiety symptoms (β = .46 STAI-state score [95% CI: 0.03-0.88] per μmol/L) adjusting for covariates. Plasma betaine concentrations were not associated with antenatal depression or anxiety symptoms. No associations were observed between pregnancy choline or betaine and postnatal mental well-being. This study suggests that higher maternal plasma choline status during pregnancy is associated with more symptoms of antenatal depression and anxiety, whereas plasma betaine concentrations showed no associations. No associations were observed for postnatal mental well-being. Prospective studies are required to replicate these findings and further examine the direction of causality and possible biological mechanisms.

Dietary Choline Intake Is Directly Associated with Bone Mineral Density in the Hordaland Health Study

BackgroundCholine is an important nutrient either obtained from a variety of foods or synthesized endogenously, and it is the precursor of betaine. We previously reported positive associations between plasma free choline and bone mineral density (BMD). Animal studies suggest an impact of dietary choline on bone metabolism, but the role of dietary intake of choline and betaine for human bone health is unknown. ObjectiveThe main aims were to examine the associations of dietary choline, choline species, and betaine with BMD and to study the relations between dietary and plasma free choline and betaine. MethodsStudy subjects were participants in the Hordaland Health Study, including 2649 women and 1983 men (aged 46–49 or 71–74 y). BMD was measured by dual-energy X-ray absorptiometry, and dietary intake was obtained by using a validated 169-item food-frequency questionnaire. Risk associations were assessed by logistic regression and correlations by ρ (Spearman's bivariate rank order correlation). ResultsSubjects in the lowest compared with the highest tertile of dietary total choline, free choline, glycerophosphocholine, phosphocholine, phosphatidylcholine, and sphingomyelin had a higher risk of low–femoral neck BMD, defined as the lowest BMD quintile. Particularly strong associations were found among middle-aged men for intake of free choline (OR: 1.83; 95% CI: 1.24, 2.69; P = 0.002) and glycerophosphocholine (OR: 2.13; 95% CI: 1.43, 3.16; P < 0.001) and among elderly women for total choline (OR: 1.96; 95% CI: 1.33, 2.88; P = 0.001) and phosphatidylcholine (OR: 1.94; 95% CI: 1.33, 2.84: P = 0.001) intake. No significant associations were observed between dietary betaine and BMD. Dietary total choline, free choline, glycerophosphocholine, phosphatidylcholine, and sphingomyelin correlated weakly with plasma free choline (ρ: 0.07, 0.05, 0.07, 0.07, and 0.05, respectively; P < 0.01). Dietary betaine correlated with plasma betaine (ρ: 0.23; P < 0.001). ConclusionsDietary choline was positively associated with BMD in middle-aged and elderly participants.

Plasma Metabolites Predict Severity of Depression and Suicidal Ideation in Psychiatric Patients-A Multicenter Pilot Analysis.

Evaluating the severity of depression (SOD), especially suicidal ideation (SI), is crucial in the treatment of not only patients with mood disorders but also psychiatric patients in general. SOD has been assessed on interviews such as the Hamilton Rating Scale for Depression (HAMD)-17, and/or self-administered questionnaires such as the Patient Health Questionnaire (PHQ)-9. However, these evaluation systems have relied on a person’s subjective information, which sometimes lead to difficulties in clinical settings. To resolve this limitation, a more objective SOD evaluation system is needed. Herein, we collected clinical data including HAMD-17/PHQ-9 and blood plasma of psychiatric patients from three independent clinical centers. We performed metabolome analysis of blood plasma using liquid chromatography mass spectrometry (LC-MS), and 123 metabolites were detected. Interestingly, five plasma metabolites (3-hydroxybutyrate (3HB), betaine, citrate, creatinine, and gamma-aminobutyric acid (GABA)) are commonly associated with SOD in all three independent cohort sets regardless of the presence or absence of medication and diagnostic difference. In addition, we have shown several metabolites are independently associated with sub-symptoms of depression including SI. We successfully created a classification model to discriminate depressive patients with or without SI by machine learning technique. Finally, we produced a pilot algorithm to predict a grade of SI with citrate and kynurenine. The above metabolites may have strongly been associated with the underlying novel biological pathophysiology of SOD. We should explore the biological impact of these metabolites on depressive symptoms by utilizing a cross species study model with human and rodents. The present multicenter pilot study offers a potential utility for measuring blood metabolites as a novel objective tool for not only assessing SOD but also evaluating therapeutic efficacy in clinical practice. In addition, modification of these metabolites by diet and/or medications may be a novel therapeutic target for depression. To clarify these aspects, clinical trials measuring metabolites before/after interventions should be conducted. Larger cohort studies including non-clinical subjects are also warranted to clarify our pilot findings.

Prospective associations of maternal betaine status with offspring weight and body composition at birth: the Growing Up in Singapore Towards healthy Outcomes (GUSTO) cohort study

Betaine supplementation results in lower body weight and fat mass and higher lean mass in animals and adult humans. However, the relation between maternal betaine status and offspring birth weight and body composition is less known. The aim was to examine the association between maternal betaine status and neonatal birth size and adiposity in an Asian mother-offspring cohort. We included 955 pregnant women whose plasma betaine concentrations were measured at 26-28 wk of gestation. Neonatal anthropometric values were measured at birth, and abdominal adipose tissue compartments were assessed by MRI in a subset of infants (n = 307) in the first 14 d after birth. Multivariate general linear models were used to adjust for gestational age; fetal sex; and maternal age, height, education, ethnicity, prepregnancy body mass index, and plasma folate, vitamin B-12, and choline concentrations. The mean ± SD plasma concentration of betaine was 13.2 ± 2.7 μmol/L (range: 5.3-25.0 μmol/L). After adjustment for covariates, higher maternal plasma betaine was associated with lower birth weight (β: -57.6 g; 95% CI: -109.9, -5.3 g), shorter birth length (β: -0.29 cm per 5-μmol/L increment; 95% CI: -0.55, -0.03 cm), smaller head circumference (-0.20 cm; 95% CI: -0.38, -0.02 cm), smaller midupper arm circumference (-0.16 cm; 95% CI: -0.30, -0.03 cm), lower volumes of abdominal superficial subcutaneous adipose tissue (-4.53 mL; 95% CI: -8.70, -0.36 mL), and a higher risk of small-for-gestational-age birth (OR: 1.57; 95% CI: 1.05, 2.35). Higher maternal betaine status was generally associated with smaller infant birth size and less abdominal fat mass. Further studies are needed to replicate these findings and to understand their biological mechanisms. This study was registered at clinicaltrials.gov as NCT01174875.