Plasma Choline Research Articles

Using Differential Threshold Effects of Individual and Combined Periconceptional Methyl Donor Status on Maternal Genomic LINE-1 and Imprinted H19 DNA Methylation to Predict Birth Weight Variance in the Taiwan Pregnancy-Newborn Epigenetics (TPNE) Cohort Study

Few studies have comprehensively examined the effect of methyl donor status on maternal DNA methylation and birth outcomes. This study examined associations between periconceptional methyl donor status and genome-wide and specific imprinted gene methylation and fetal growth indices in the Taiwan Pregnancy-Newborn Epigenetics cohort. Plasma folate, choline (free form), and betaine concentrations of the participants enrolled at 7-10 weeks of gestation were analyzed. DNA methylation at regulatory sequences of the imprinted H19 gene and genomic long interspersed nuclear element 1 (LINE-1) were measured in maternal lymphocytes using bisulfite/high-resolution melt polymerase chain reaction. Associations with birth weight (BW) were estimated through multiple regressions from 112 mother-newborn pairs. A nonlinear "L-shaped" relation and an inverse association between maternal plasma folate in T1 (mean ± SE: 17.6±5.1 nmol/L) and lymphocytic LINE-1 methylation (β:-0.49, P=0.027) were characterized. After adjusting for LINE-1 methylation, individual maternal folate concentrations were positively associated with BW variance (β=0.24, P=0.035), and the association was more pronounced in mothers with choline in T1 (mean ± SE: 5.4±0.6 μmol/L; β: 0.40, P=0.039). Choline status of the mothers in T2 (mean ± SE: 7.2±0.6 μmol/L) was inversely associated with LINE-1 methylation (β: -0.43, P=0.035), and a positive association was evident between T1 choline and H19 methylation (β: 0.48, P=0.011). After adjusting for epigenetic modification, maternal choline status predicted a positive association with BW (β: 0.56, P=0.005), but the effect was limited to mothers with high betaine concentrations in T3 (mean ± SE: 36.4±8.8 μmol/L), depending on folate status. Our data highlight the differential threshold effects of periconceptional folate, choline, and betaine status on genomic LINE-1 and H19 DNA methylation and how their interplay has a long-term effect on BW variance.

Quantitative metabolic biomarker analysis of mild cognitive impairment in eastern U.P. and Bihar population

Mild cognitive impairment (MCI) is a transition phase between healthy individuals and Alzheimer’s disease (AD). Therefore, diagnosis of MCI at early stage will help to delay or prevent its progression to disease. In the present study, we aim to identify the metabolic biomarkers, which can help in the diagnosis of MCI. We have screened 2000 elderly individuals from north India, out of which 200 were identified as MCI. We continued our study on 10 MCI individuals who regularly participated in the follow-up. The age and gender matched 10 healthy individuals were taken as control. These control and MCI individuals were subjected to neuropsychological examination such as Hindi mental state examination (HMSE) and Montreal cognitive assessment (MOCA) followed by 1H Nuclear Magnetic Resonance (NMR) analysis. Remarkable changes were noted between control and MCI individuals at metabolic level. In silico study showed the involvement of eight metabolites in MCI. We found higher level of lactate, N-acetyl aspartate, histidine and lower level of formate, choline, alanine, creatinine and glucose in blood plasma of MCI individuals compared to control. Further, In silico study showed that choline might be directly associated with MCI or AD. Such In silico study with quantitative metabolite analysis of plasma could be used as diagnostic biomarkers for the identification of MCI.

54 Choline alters the pattern of DNA methylation and lipid content of pre-implantation bovine embryos

Choline is a methyl donor for DNA methylation and a precursor for phosphatidylcholine, which is the major phospholipid of cell membranes. Early embryonic development involves processes of DNA demethylation and remethylation as well as synthesis of new cell membranes. Addition of choline chloride (ChCl) to culture medium of embryos produced invitro increased birthweight of Brahman calves after embryo transfer. The objective was to determine whether the addition of ChCl to culture medium alters the pattern of DNA methylation and lipid content of pre-implantation embryos produced invitro. Embryos were incubated after fertilisation in BBH7 culture medium containing 0.0, 0.004, 1.3, or 1.8mM ChCl (adjusted with NaCl to maintain isotonicity). Concentrations were chosen to approximate free choline (0.004mM) and total choline (1.30mM) in plasma of cows at week 1 postpartum and concentration of total choline in plasma of cows fed rumen-protected choline (1.8mM). Cleavage and blastocyst rate (n=8 replicates) were evaluated at Days 3 and 7.5 post-insemination, respectively. Embryos ≥8 cells (range 8 to 24 cells; stages near embryonic genome activation) and expanded blastocysts were harvested at Day 3.75 (n=232) and 7.5 (n=204) to estimate global DNA methylation by immunostaining for 5-methyl-cytosine. Methylation of DNA was estimated by calculating the ratio of fluorescence for 5-methyl-cytosine to that of propidium iodide (DNA). Another group of expanded blastocysts was harvested (n=99) to estimate lipid content using Nile Red. Embryo development was analysed by GLIMMIX procedure and fluorescence data by GLM procedure of SAS (SAS Institute Inc.). The proportion of zygotes that cleaved after fertilisation (77.5±2.3, 78.1±2.3, 74.5±2.4, and 80.1±2.2% for 0.0, 0.004, 1.3, and 1.8mM ChCl; P=0.2736) and cleaved embryos that became blastocysts (37.8±4.4, 41.5±4.6, 42.8±4.6, and 39.6±4.4%; P=0.5764) was similar between treatments. The DNA methylation at both days was affected by treatment (P<0.001). At Day 3.75, 1.3mM choline reduced methylation and there were no effects of other concentrations (1.13a±0.03, 1.04a±0.03, 0.92b±0.03, and 1.13a±0.04; means with different superscripts differ at P<0.05). For blastocysts, in contrast, DNA methylation was increased for embryos treated with 1.3 and 1.8mM choline (0.98a±0.04, 1.04ab±0.03, 1.25c±0.03, and 1.11b±0.03). Lipid content in blastocysts was also affected by treatment (P=0.0139). In particular, lipid content was higher for embryos treated with 1.3 and 1.8mM choline (409.1a±54.3, 542.3ab±62.3, 651.3b±54.3, and 583.9b±55.0). In conclusion, addition of ChCl to culture medium altered DNA methylation in bovine pre-implantation embryos produced invitro in a manner dependent on developmental stage and choline concentration. Likewise, ChCl increased lipid content in the resultant blastocysts. Support was provided by the Red Larson Endowment.

Differences in response to egg-derived dietary cholesterol result in distinct lipoprotein profiles while plasma concentrations of carotenoids and choline are not affected in a young healthy population

Abstract Background It has been demonstrated that some individuals (approximately 1/3 of the population) can be classified as hyper-responders to dietary cholesterol. Objective The objective of this study was to evaluate whether the response to dietary cholesterol (provided by eggs) would mediate lipoprotein metabolism and the bioavailability of lutein, zeaxanthin, and choline, nutritional components present in eggs. Methods We recruited 36 healthy subjects who underwent a two-week washout period in which they consumed zero eggs. Participants were then consecutively fed 1, 2, and 3 eggs/day for 4 weeks each. In this analysis, we compared intake of 0 eggs/day to 3 eggs/day (540 additional mg of dietary cholesterol/day). Plasma lipids, lipoprotein particle subfractions, and plasma lutein, zeaxanthin, and choline were measured. We classified the normal plasma response to dietary cholesterol as an increase of 2.2 mg/dL for each 100 mg of dietary cholesterol/day. Therefore, those individuals with >12 mg/dL increase in plasma cholesterol after the intervention (n = 12) were considered hyper-responders while the remaining 24 subjects presented changes in plasma cholesterol ranging from −16 to 9 mg/dL and were considered normal responders. Results Compared to 0 eggs, hyper-responders had significant increases in plasma total, LDL and HDL cholesterol (p Conclusions These results suggest that egg intake increases the bioavailability of carotenoids and choline present in eggs independently of individual responses to dietary cholesterol. Further, the observed increases in large HDL in all individuals could be associated with HDL being a major transporter of lutein and zeaxanthin in plasma.

Natural Choline from Egg Yolk Phospholipids Is More Efficiently Absorbed Compared with Choline Bitartrate; Outcomes of A Randomized Trial in Healthy Adults.

Choline is a vitamin-like essential nutrient, important throughout one’s lifespan. Therefore, choline salts are added to infant formula, supplements and functional foods. However, if choline is present in a natural form, e.g. bound to phospholipids, it may be more efficiently absorbed. The study’s aim was to evaluate if choline uptake is improved after consumption of an egg yolk phospholipid drink, containing 3 g of phospholipid bound choline, compared to a control drink with 3 g of choline bitartrate. We performed a randomized, double blind, cross-over trial with 18 participants. Plasma choline, betaine and dimethylglycine concentrations were determined before and up to six hours after consumption of the drinks. The plasma choline response, as determined by the incremental area under the curve, was four times higher after consumption of the egg yolk phospholipid drink compared with the control drink (p < 0.01). Similar outcomes were also observed for choline’s main metabolites, betaine (p < 0.01) and dimethylglycine (p = 0.01). Consumption of natural choline from egg yolk phospholipids improved choline absorption compared to consumption of chemically produced choline bitartrate. This information is of relevance for the food industry, instead of adding choline-salts, adding choline from egg yolk phospholipids can improve choline uptake and positively impact health.

Dietary choline is related to increased risk of acute myocardial infarction in patients with stable angina pectoris

High plasma choline has been associated with the metabolic syndrome and risk of chronic diseases, including cardiovascular disease. However, dietary choline is not correlated with choline plasma concentrations, and there are few studies and contradictory evidence regarding dietary choline and cardiovascular events. In addition, a recommended dietary allowance for choline has not been established and remains a point of contention.This study assessed the association between dietary choline, including choline forms, and risk of incident acute myocardial infarction (AMI) in patients with suspected stable angina pectoris (SAP).In total 1981 patients (80% men, median age 62) from the Western Norway B Vitamin Intervention Trial were included in this analysis. Information on dietary choline was obtained using a 169-item food frequency questionnaire. The Cardiovascular Disease in Norway project provided data on AMI. Risk associations were estimated using Cox-regression analysis using energy-adjusted choline intake.Median (25th, 75th percentile) total energy-adjusted choline intake was 288 (255, 326) mg/d. During a median (25th, 75th percentile) follow-up of 7.5 (6.3, 8.8) years, 312 (15.7%) patients experienced at least one AMI. Increased intakes of energy-adjusted choline (HR [95% CI] per 50 mg increase 1.11 [1.03, 1.20]), phosphatidylcholine (HR per 50 mg increase 1.24 [1.08, 1.42]) and sphingomyelin (HR per 5 mg increase 1.16 [1.02, 1.31]) were associated with higher AMI risk.In conclusion, higher dietary intakes of total choline, phosphatidylcholine and sphingomyelin were associated with increased risk of AMI in patients with SAP. Future studies are necessary to explore underlying mechanisms for this observation.

P1531The association between plasma choline and acute myocardial infarction is modified by potential markers of endogenous PPAR activation

Abstract Background Choline is related to lipid handling and higher plasma concentrations have been associated with an adverse cardiovascular risk profile. However, previous studies have suggested that the relationship between plasma free choline and later cardiovascular events may differ according to patient phenotypes. Purpose To explore the risk association between plasma choline and later acute myocardial infarction (AMI) according to plasma methylmalonic acid (MMA) or dimethylglycine (DMG). The latter two metabolites are suggested markers of endogenous activation of perixosome proliferator-activated receptors (PPARs), which are nuclear receptor proteins involved in lipid metabolism. Methods Risk relationships were explored by Cox regression among 2232 patients evaluated for suspected stable angina pectoris in the overall population and according to median plasma MMA and DMG. Results Baseline plasma choline was related to several cardiovascular risk factors (Table 1). After median follow-up of 7.3 years, 338 patients were reported with at least one incident AMI. In the overall population, the age and gender adjusted HR (95% CI) for each increment of 1 SD log-transformed plasma choline and AMI was 1.21 (1.08–1.35), P=0.001, and the association persisted in multivariate analyses. In patients with plasma MMA or DMG≥median, the HRs (95% CIs) were 1.33 (1.16–1.54) and 1.38 (1.20–1.58), respectively, both P&lt;0.0001; however no significant relationships were observed between plasma choline and later AMI among patients with either plasma MMA or DMG &lt; median (P interaction &lt;0.008) (Figure 1). &lt;MEDIAN (P&gt; Table 1. Baseline characteristics according to plasma choline quartiles Quartile 1 Quartile 4 P for trend Age, years 58 (52–66) 66 (58–73) &lt;0.0001 Smoking, n (%) 212 (37.9) 153 (27.9) &lt;0.0001 Diabetes, n (%) 61 (10.9) 85 (15.5) 0.12 Previous acute myocardial infarction, n (%) 200 (35.7) 238 (43.4) &lt;0.0001 Estimated glomerular filtration rate, mL/min/1.73m2 96 (87–104) 79 (63–92) &lt;0.0001 Serum hs-troponin T, ng/L 4 (3–8) 9 (4–17) 0.0002 Serum triglycerides, mmol/L 1.35 (1.00–2.03) 1.60 (1.16–2.25) &lt;0.0001 Serum apolipoprotein A1, mg/L 1.29 (1.12–1.51) 1.32 (1.17–1.53) 0.01 Statin therapy, n (%) 384 (68.6) 435 (79.4) 0.01 Figure 1 Conclusion Among patients with stable angina, plasma choline was related to increased long-term AMI risk among patients with higher plasma MMA or DMG only. This finding potentially reflects increased risk conferred by choline during concomitant endogenous PPAR activation. Acknowledgement/Funding None

Hepatic betaine-homocysteine methyltransferase and methionine synthase activity and intermediates of the methionine cycle are altered by choline supply during negative energy balance in Holstein cows

Although choline requirements are unknown, enhanced postruminal supply may decrease liver triacylglycerol (TAG) storage and increase flux through the methionine cycle, helping cows during a negative energy balance (NEB). The objective was to investigate effects of postruminal choline supply during NEB on hepatic activity of betaine-homocysteine methyltransferase (BHMT), methionine synthase (MTR), methionine adenosyltransferase, transcription of enzymes, and metabolite concentrations in the methionine cycle. Ten primiparous rumen-cannulated Holstein cows (158 ± 24 d postpartum) were used in a replicated 5 × 5 Latin square design with 4-d treatment periods and 10 d of recovery (14 d/period). Treatments were unrestricted intake with abomasal infusion of water (A0), restricted intake (R; 60% of net energy for lactation requirements to induce NEB) with abomasal infusion of water (R0) or R plus abomasal infusion of 6.25, 12.5, or 25 g/d of choline ion. Liver tissue was collected on d 5 after the infusions ended, blood on d 1 to 5, and milk on d 1 to 4. Statistical contrasts were A0 versus R0 (CONT1) and tests of linear (L), quadratic (Q), and cubic (C) effects of choline dose. Plasma choline increased with R (CONT1) and choline (L). Although R decreased milk yield (CONT1), choline increased milk yield and liver phosphatidylcholine (PC), but decreased TAG (L). No differences were observed in plasma PC or very-low-density lipoprotein concentrations with R or choline. Activity and mRNA abundance of BHMT were greater with R (CONT1) and increased with choline (L). Although activity of MTR was lower with R (CONT1), it tended to increase with choline (L). No effect of R was detected for activity of methionine adenosyltransferase, but it changed cubically across dose of choline. Those responses were associated with linear increases in the concentrations of liver tissue (+13%) and plasma methionine concentrations. The mRNA abundance of CPT1A, SLC22A5, APOA5, and APOB, genes associated with fatty acid oxidation and lipoprotein metabolism, was upregulated by choline (Q). Overall, enhanced supply of choline during NEB increases hepatic activity of BHMT and MTR to regenerate methionine and PC, partly to help clear TAG. The relevance of these effects during the periparturient period merits further research.

Quantitative monitoring of a panel of stress-induced biomarkers in human plasma by liquid chromatography-tandem mass spectrometry: an application in a comparative study between depressive patients and healthy subjects.

Using a metabolomic approach, we have found that stress can induce oxidative damage by disturbing the creatine/phosphocreatine shuttle system and purinergic pathway, leading to an excessive membrane breakdown. To further validate our findings and to monitor the biological impact of stress in research of clinical psychiatry, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed to simultaneously determine a panel of biomarkers comprising choline, creatine, purinergic metabolites, neurosteroids, lysophosphatidylcholines, and phosphatidylethanolamines in human plasma. After optimization of the extraction protocol, all the 15 analytes plus 4 internal standards with distinct polarities were extracted into an organic phase using methyl tert-butyl ether/methanol (1:1, v/v). A reversed-phase C8 column under gradient elution consisting of aqueous phase A of 5mM ammonium acetate buffer solution containing 0.1% formic acid and organic phase B of acetonitrile/2-propanol (3:7, v/v) was utilized for separation. Four sequential periods under positive or negative ion mode were combined for the determination of analytes with specific multiple reaction monitoring transitions. For all analytes, this method exhibited good linearity with coefficients of determination (R2) higher than 0.99. The lower limit of quantification (LLOQ) values ranged from 0.05 to 80.0ng/mL. Recovery between 70.5 and 97.3% was obtained by spiking standards to plasma samples stripped by powdered activated carbon. The intra- and inter-assay relative standard deviations (RSDs) of the analyses varied between 2.0 and 13.3%. The mean accuracy ranged from 90.6 to 109.0%. The matrix effect ranged from 91.2 to 107.3% with variations less than 9.0%. Stability under different conditions was tested, with mean recoveries varying between 90.4 and 109.7%. Finally, the established method was successfully applied to analyze the plasma samples from a small cohort of 30 patients with major depressive disorder and 30 matched healthy controls. Graphical abstract.

Differences in Response to Egg Intake Result in Distinct Lipoprotein Profiles While Plasma Concentrations of Carotenoids and Choline Are Not Affected (P08-102-19)

ObjectivesIt has been demonstrated that some individuals (approximately 1/3 of the population) can be classified as hyper-responders to dietary cholesterol. The objective of this study was to evaluate whether the response to dietary cholesterol (provided by eggs) would affect lipoprotein metabolism and the bioavailability of lutein, zeaxanthin and choline, nutritional components present in eggs. MethodsWe recruited 36 healthy subjects who underwent a two-week washout period in which they consumed zero eggs. Participants were then fed 3 eggs (540 additional mg of dietary cholesterol) for 4 weeks. Plasma lipids were measured by use of an automated chemical clinical analyzer; the number of large and small LDL and HDL particles were measured by Nuclear Magnetic Resonance. Plasma lutein and zeaxanthin were determined by use of HPLC and plasma choline by use of LSM spectroscopy. We classified the normal response to dietary cholesterol as 2.2 mg/dL for each additional 100 mg of dietary cholesterol. Therefore, those individuals who had >12 mg/dL increase in plasma cholesterol after the intervention (n = 12) were considered hyper-responders while the other 24 subjects who presented changes in plasma cholesterol ranging from −16 to 9 mg/dL were considered hypo-responders. ResultsCompared to zero eggs, hyper-responders had significant increases in plasma total, LDL and HDL cholesterol (P < 0.001) with no changes in the LDL/HDL ratio. They also had higher concentrations of large LDL (P < 0.01) with no changes in small LDL. In contrast no changes in plasma total LDL or HDL cholesterol or large LDL were observed in hypo-responders after the consumption of 3 eggs/day. Interestingly, both hypo and hyper-responders had significant increases in large HDL particles (12%), plasma lutein (17%), zeaxanthin (30%) and choline (12%) (P < 0.001) compared to zero egg intake. ConclusionsThese results suggest that egg intake increases the bioavailability of carotenoids and choline present in eggs independently of plasma cholesterol response to dietary cholesterol. Further, the observed increases in large HDL in all individuals could be associated with HDL being a major transporter of lutein and zeaxanthin in plasma. Funding SourcesThe Esperance Family and the Egg Nutrition Center.

Periconceptional alcohol exposure causes female-specific perturbations to trophoblast differentiation and placental formation in the rat.

The development of pathologies during pregnancy, including pre-eclampsia, hypertension and fetal growth restriction (FGR), often originates from poor functioning of the placenta. In vivo models of maternal stressors, such as nutrient deficiency, and placental insufficiency often focus on inadequate growth of the fetus and placenta in late gestation. These studies rarely investigate the origins of poor placental formation in early gestation, including those affecting the pre-implantation embryo and/or the uterine environment. The current study characterises the impact on blastocyst, uterine and placental outcomes in a rat model of periconceptional alcohol exposure, in which 12.5% ethanol is administered in a liquid diet from 4 days before until 4 days after conception. We show female-specific effects on trophoblast differentiation, embryo-uterine communication, and formation of the placental vasculature, resulting in markedly reduced placental volume at embryonic day 15. Both sexes exhibited reduced trophectoderm pluripotency and global hypermethylation, suggestive of inappropriate epigenetic reprogramming. Furthermore, evidence of reduced placental nutrient exchange and reduced pre-implantation maternal plasma choline levels offers significant mechanistic insight into the origins of FGR in this model.

Phosphatidylcholine from krill increases plasma choline and its metabolites in dogs.

Background and Aim:Choline and its metabolites have multiple physiological roles in the body, which are important for muscle function, memory, methylation reactions, and hepatic lipid transport. This study aimed to investigate, if inclusion of phosphatidylcholine (PC) from Antarctic krill (Euphausia superba) can increase the concentration of choline and its metabolites in plasma of sled dogs in comparison to a control group.Materials and Methods:Ten adult Alaskan Huskies of both genders were supplemented with PC from 8% dietary krill meal inclusion for 6 weeks, while another ten dogs received no krill meal supplementation. Blood measurements of the two groups were taken at baseline and end of the study and compared for choline and its metabolite concentrations.Results:The choline concentration of the krill meal-supplemented dogs was significantly higher after 6 weeks of krill meal feeding compared to the control group (mean increase = 4.53 µmol/L in the supplemented versus 1.21 µmol/L in the control group, p=0.014). Furthermore, krill meal-supplemented dogs showed significantly more pronounced increases in betaine (p<0.001), dimethylglycine (p<0.01), trimethylamine-N-oxide (p<0.001), and trimethyllysine (p<0.001) compared to the control group. Significant correlations between changes in choline and changes in its metabolites were observed.Conclusion:The results showed that krill meal supplementation was associated with significantly higher plasma choline concentrations, which correlated with changed concentrations of choline metabolites.

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.

Choline Supplementation in Cystic Fibrosis-The Metabolic and Clinical Impact.

Background: Choline is essential for the synthesis of liver phosphatidylcholine (PC), parenchymal maintenance, bile formation, and lipoprotein assembly to secrete triglycerides. In choline deficiency, the liver accretes choline/PC at the expense of lung tissue, thereby impairing pulmonary PC homoeostasis. In cystic fibrosis (CF), exocrine pancreas insufficiency results in impaired cleavage of bile PC and subsequent fecal choline loss. In these patients, the plasma choline concentration is low and correlates with lung function. We therefore investigated the effect of choline supplementation on plasma choline/PC concentration and metabolism, lung function, and liver fat. Methods: 10 adult male CF patients were recruited (11/2014–1/2016), and orally supplemented with 3 × 1 g choline chloride for 84 (84–91) days. Pre-/post-supplementation, patients were spiked with 3.6 mg/kg [methyl-D9]choline chloride to assess choline/PC metabolism. Mass spectrometry, spirometry, and hepatic nuclear resonance spectrometry served for analysis. Results: Supplementation increased plasma choline from 4.8 (4.1–6.2) µmol/L to 10.5 (8.5–15.5) µmol/L at d84 (p < 0.01). Whereas plasma PC concentration remained unchanged, D9-labeled PC was decreased (12.2 [10.5–18.3] µmol/L vs. 17.7 [15.5–22.4] µmol/L, p < 0.01), indicating D9-tracer dilution due to higher choline pools. Supplementation increased Forced Expiratory Volume in 1 second percent of predicted (ppFEV1) from 70.0 (50.9–74.8)% to 78.3 (60.1–83.9)% (p < 0.05), and decreased liver fat from 1.58 (0.37–8.82)% to 0.84 (0.56–1.17)% (p < 0.01). Plasma choline returned to baseline concentration within 60 h. Conclusions: Choline supplementation normalized plasma choline concentration and increased choline-containing PC precursor pools in adult CF patients. Improved lung function and decreased liver fat suggest that in CF correcting choline deficiency is clinically important. Choline supplementation of CF patients should be further investigated in randomized, placebo-controlled trials.

Plasma TMAO and Choline Levels in Individuals With Metabolic Syndrome - Comparison Between Eggs and Choline Supplement.

Plasma TMAO and Choline Levels in Individuals With Metabolic Syndrome - Comparison Between Eggs and Choline Supplement.

Combined choline and DHA supplementation: a randomized controlled trial.

Choline and docosahexaenoic acid (DHA) are essential nutrients for preterm infant development. They are metabolically linked via phosphatidylcholine (PC), a constitutive plasma membrane lipid and the major transport form of DHA in plasma. Plasma choline and DHA-PC concentrations rapidly decline after preterm birth. To improve preterm infant nutrition, we evaluated combined compared to exclusive choline and DHA supplementation, and standard feeding. Randomized partially blinded single-center trial. Neonatal tertiary referral center in Tübingen, Germany. 24 inborn preterm infants < 32 week postmenstrual age. Standard nutrition (control) or, additionally, enteral choline (30mg/kg/day), DHA (60mg/kg/day), or both for 10days. Single enteral administration of 3.6mg/kg [methyl-D9-] choline chloride as a tracer at 7.5days. Primary outcome variable was plasma choline following 7 days of supplementation. Deuterated and unlabeled choline metabolites, DHA-PC, and other PC species were secondary outcome variables. Choline supplementation increased plasma choline to near-fetal concentrations [35.4 (32.8-41.7)µmol/L vs. 17.8 (16.1-22.4) µmol/L, p < 0.01] and decreased D9-choline enrichment of PC. Single DHA treatment decreased DHA in PC relative to total lipid [66 (60-68)% vs. 78 (74-80)%; p < 0.01], which was prevented by choline. DHA alone increased DHA-PC only by 35 (26-45)%, but combined treatment by 63 (49-74)% (p < 0.001). D9-choline enrichment showed preferential synthesis of PC containing linoleic acid. PC synthesis via phosphatidylethanolamine methylation resulted in preferential synthesis of DHA-containing D3-PC, which was increased by choline supplementation. 30mg/kg/day additional choline supplementation increases plasma choline to near-fetal concentrations, dilutes the D9-choline tracer via increased precursor concentrations and improves DHA homeostasis in preterm infants. clinicaltrials.gov. Identifier: NCT02509728.

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.

Choline and choline-related nutrients in regular and preterm infant growth.

Choline is an essential nutrient, with increased requirements during development. It forms the headgroup of phosphatidylcholine and sphingomyelin in all membranes and many secretions. Phosphatidylcholine is linked to cell signaling as a phosphocholine donor to synthesize sphingomyelin from ceramide, a trigger of apoptosis, and is the major carrier of arachidonic and docosahexaenoic acid in plasma. Acetylcholine is important for neurodevelopment and the placental storage form for fetal choline supply. Betaine, a choline metabolite, functions as osmolyte and methyl donor. Their concentrations are all tightly regulated in tissues. During the fetal growth spurt at 24-34-week postmenstrual age, plasma choline is higher than beyond 34 weeks, and threefold higher than in pregnant women [45 (36-60)µmol/L vs. 14 (10-17)µmol/L]. The rapid decrease in plasma choline after premature birth suggests an untimely reduction in choline supply, as cellular uptake is proportional to plasma concentration. Supply via breast milk, with phosphocholine and α-glycerophosphocholine as its major choline components, does not prevent such postnatal decrease. Moreover, high amounts of liver PC are secreted via bile, causing rapid hepatic choline turnover via the enterohepatic cycle, and deficiency in case of pancreatic phospholipase A2 deficiency or intestinal resection. Choline deficiency causes hepatic damage and choline accretion at the expense of the lungs and other tissues. Choline deficiency may contribute to the impaired lean body mass growth and pulmonary and neurocognitive development of preterm infants despite adequate macronutrient supply and weight gain. In this context, a reconsideration of current recommendations for choline supply to preterm infants is required.

Hepatic Expression of PEMT, but Not Dietary Choline Supplementation, Reverses the Protection against Atherosclerosis in Pemt−/−/Ldlr−/− Mice

Phosphatidylethanolamine N-methyltransferase (PEMT) converts phosphatidylethanolamine to phosphatidylcholine. Pemt-/-/low density lipoprotein receptor (Ldlr)-/- mice have significantly reduced plasma lipids and are protected against atherosclerosis. Recent studies have shown that choline can be metabolized by the gut flora into trimethylamine-N-oxide (TMAO), which is an emerging risk factor for atherosclerosis. The objective of this study was to determine whether ectopic hepatic PEMT expression or choline supplementation would promote atherosclerosis in Pemt-/-/Ldlr-/- mice. Male 8- to 10-wk-old Pemt+/+/Ldlr-/- (SKO) and Pemt-/-/Ldlr-/- (DKO) mice were injected with an adeno-associated virus (AAV) expressing green fluorescent protein (GFP) or human PEMT and fed a Western diet (40% of calories from fat, 0.5% cholesterol) for 8 wk. In a separate experiment, 8- to 10-wk-old SKO and half of the DKO male mice were fed a Western diet with normal (3 g/kg) choline for 12 wk. The remaining DKO mice [choline-supplemented (CS) DKO] were fed a CS Western diet (10 g choline/kg). Plasma lipid concentrations, choline metabolites, and aortic atherosclerosis were measured. Plasma cholesterol, plasma TMAO, and aortic atherosclerosis were reduced by 60%, 40%, and 80%, respectively, in DKO mice compared with SKO mice. AAV-PEMT administration increased plasma cholesterol and TMAO by 30% and 40%, respectively, in DKO mice compared with AAV-GFP-treated DKO mice. Furthermore, AAV-PEMT-injected DKO mice developed atherosclerotic lesions similar to SKO mice. In the second study, there was no difference in atherosclerosis or plasma cholesterol between DKO and CS-DKO mice. However, plasma TMAO concentrations were increased 2.5-fold in CS-DKO mice compared with DKO mice. Reintroducing hepatic PEMT reversed the atheroprotective phenotype of DKO mice. Choline supplementation did not increase atherosclerosis or plasma cholesterol in DKO mice. Our data suggest that plasma TMAO does not induce atherosclerosis when plasma cholesterol is low. Furthermore, this is the first report to our knowledge that suggests that de novo choline synthesis alters TMAO status.

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.