Choline Status Research Articles

Assessing the Benefit of Dietary Choline Supplementation Throughout Adulthood in the Ts65Dn Mouse Model of Down Syndrome

Background/Objectives: Down syndrome (DS) is the most common cause of early-onset Alzheimer’s disease (AD). Dietary choline has been proposed as a modifiable factor to improve the cognitive and pathological outcomes of AD and DS, especially as many do not reach adequate daily intake levels of choline. While lower circulating choline levels correlate with worse pathological measures in AD patients, choline status and intake in DS is widely understudied. Perinatal choline supplementation (Ch+) in the Ts65Dn mouse model of DS protects offspring against AD-relevant pathology and improves cognition. Further, dietary Ch+ in adult AD models also ameliorates pathology and improves cognition. However, dietary Ch+ in adult Ts65Dn mice has not yet been explored; thus, this study aimed to supply Ch+ throughout adulthood to determine the effects on cognition and DS co-morbidities. Methods: We fed trisomic Ts65Dn mice and disomic littermate controls either a choline normal (ChN; 1.1 g/kg) or a Ch+ (5 g/kg) diet from 4.5 to 14 months of age. Results: We found that Ch+ in adulthood failed to improve genotype-specific deficits in spatial learning. However, in both genotypes of female mice, Ch+ significantly improved cognitive flexibility in a reverse place preference task in the IntelliCage behavioral phenotyping system. Further, Ch+ significantly reduced weight gain and peripheral inflammation in female mice of both genotypes, and significantly improved glucose metabolism in male mice of both genotypes. Conclusions: Our findings suggest that adulthood choline supplementation benefits behavioral and biological factors important for general well-being in DS and related to AD risk.

A New, Accurate LC-MS/MS Method for Assessment of the Choline Status of the US Population

A New, Accurate LC-MS/MS Method for Assessment of the Choline Status of the US Population

Is choline deficiency an unrecognized factor in necrotizing enterocolitis of preterm infants?

We undertook this review to determine if it is plausible that choline or phosphatidylcholine (PC) deficiency is a factor in necrotizing enterocolitis (NEC) after two clinical trials found a dramatic and unexpected reduction in NEC in an experimental group provided higher PC compared to a control group. Sources and amounts of choline/PC for preterm infants are compared to the choline status of preterm infants at birth and following conventional nutritional management. The roles of choline/PC in intestinal structure, mucus, mesenteric blood flow, and the cholinergic anti-inflammatory system are summarized. Low choline/PC status is linked to prematurity/immaturity, parenteral and enteral feeding, microbial dysbiosis and hypoxia/ischemia, factors long associated with the risk of developing NEC. We conclude that low choline status exists in preterm infants provided conventional parenteral and enteral nutritional management, and that it is plausible low choline/PC status adversely affects intestinal function to set up the vicious cycle of inflammation, loss of intestinal barrier function and worsening tissue hypoxia that occurs with NEC. In conclusion, this review supports the need for randomized clinical trials to test the hypothesis that additional choline or PC provided parenterally or enterally can reduce the incidence of NEC in preterm infants. IMPACT STATEMENT: Low choline status in preterm infants who are managed by conventional nutrition is plausibly linked to the risk of developing necrotizing enterocolitis.

Polymorphisms in the choline transporter SLC44A1 are associated with reduced cognitive performance in normotypic but not prenatal alcohol-exposed children

BackgroundCholine is essential for healthy cognitive development. Single nucleotide polymorphisms (SNPs; rs3199966(G), rs2771040(G)) within the choline transporter SLC44A1 increase risk for choline deficiency. In a choline intervention trial of children who experienced prenatal alcohol exposure (PAE), these alleles are associated with improved cognition. ObjectiveThis study aimed to determine if SNPs within SLC44A1 are differentially associated with cognition in children with PAE compared with normotypic controls (genotype × exposure). A secondary objective tested for an association of these SNPs and cognition in controls (genotype-only). DesignThis is a secondary analysis of data from the Collaborative Initiative on Fetal Alcohol Spectrum Disorders. Participants (163 normotypic controls, 162 PAE) underwent psychological assessments and were genotyped within SLC44A1. Choline status was not assessed. Association analysis between genotype × exposure was performed using an additive genetic model and linear regression to identify the allelic effect. The primary outcome was the interaction between SLC44A1 genotype × exposure status with respect to cognition. The secondary outcome was the cognitive–genotype association in normotypic controls. ResultsGenotype × exposure analysis identified 7 SNPs in SLC44A1, including rs3199966(G) and rs2771040(G), and in strong linkage (D′ ≥ 0.87), that were associated (adjusted P ≤ 0.05) with reduced performance in measures of general cognition, nonverbal and quantitative reasoning, memory, and executive function (β, 1.92–3.91). In controls, carriers of rs3199966(GT or GG) had worsened cognitive performance than rs3199966(TT) carriers (β, 0.46–0.83; P < 0.0001), whereas cognitive performance did not differ by rs3199966 genotype in those with PAE. ConclusionsTwo functional alleles that increase vulnerability to choline deficiency, rs3199966(G) (Ser644Ala) and rs2771040(G) (3′ untranslated region), are associated with worsened cognition in otherwise normotypic children. These alleles were previously associated with greater cognitive improvement in children with PAE who received supplemental choline. The findings endorse that choline benefits cognitive development in normotypic children and those with PAE.

P16-041-23 Combined Oral Contraceptive Use Alters the Effects of Egg Intake on Lipoprotein Profiles,Without Impacting Effects on Insulin Resistance, Inflammation, or Choline Status

P16-041-23 Combined Oral Contraceptive Use Alters the Effects of Egg Intake on Lipoprotein Profiles,Without Impacting Effects on Insulin Resistance, Inflammation, or Choline Status

The Role of Choline in Neurodevelopmental Disorders-A Narrative Review Focusing on ASC, ADHD and Dyslexia.

Neurodevelopmental disorders appear to be rising in prevalence, according to the recent Global Burden of Disease Study. This rise is likely to be multi-factorial, but the role of certain nutrients known to facilitate neurodevelopment should be considered. One possible contributing factor could be attributed to deficits in choline intake, particularly during key stages of neurodevelopment, which includes the first 1000 days of life and childhood. Choline, a key micronutrient, is crucial for optimal neurodevelopment and brain functioning of offspring. The present narrative review discusses the main research, describing the effect of choline in neurodevelopmental disorders, to better understand its role in the etiology and management of these disorders. In terms of findings, low choline intakes and reduced or altered choline status have been reported in relevant population subgroups: pregnancy (in utero), children with autism spectrum disorders, people with attention deficit hyperactivity disorder and those with dyslexia. In conclusion, an optimal choline provision may offer some neuronal protection in early life and help to mitigate some cognitive effects in later life attributed to neurodevelopmental conditions. Research indicates that choline may act as a modifiable risk factor for certain neurodevelopmental conditions. Ongoing research is needed to unravel the mechanisms and explanations.

Choline, DHA, and Diarrheal Disease Associated with Growth Faltering in a Case-Control Study

ABSTRACTBackgroundChildren with recurrent infectious diarrhea are susceptible to growth faltering. DHA and choline may play a role in this relationship due to their involvement in lipid metabolism, gut immunity, and inflammatory pathways.ObjectivesThis study aimed to characterize the contributions made by DHA and choline status and enteric damage in young children in the association between diarrheal illness and child growth.MethodsA longitudinal case-control study was conducted among children aged 6–36 mo (N = 195) in Cap-Haitien, Haiti. Mother-child dyads were recruited from community health posts and outpatient clinics. Cases were defined as children experiencing acute diarrhea within the last 3 d and matched to healthy controls. Child anthropometry, dietary intake, and blood and stool samples were collected at baseline and follow-up. Plasma DHA, choline, and betaine were determined by LC-MS/MS methods (n = 49) and intestinal fatty acid–binding protein (I-FABP) by ELISA (n = 183). Multivariate regression models were applied with mediation analyses to examine associations and adjust for confounding factors.ResultsAt baseline, mean plasma DHA concentrations (1.03 µg/mL; 95% CI: 0.91, 1.15) were not significantly different between cases and controls, nor was there a difference in mean plasma choline concentrations (4.5 µg/mL; 95% CI: 3.8, 5.1). Mean plasma I-FABP concentrations were significantly higher at follow-up in cases (3.34; 95% CI: 3.28, 3.40) than controls (3.20; 95% CI: 3.13, 3.27; P = 0.002). In adjusted multilinear regression models, higher plasma DHA concentrations at follow-up were associated with a negative change in weight-age z score (P = 0.016), and follow-up I-FABP was inversely associated with height-age z score (P = 0.035). No interaction or mediation effects were found.ConclusionsI-FABP concentrations were significantly higher in cases as compared with controls at follow-up, suggesting ongoing enteric damage and increased risk for malnutrition. Plasma DHA and I-FABP may have a role in childhood growth outcomes.

No Association Between Riboflavin and Choline Status in a Sample of Canadian and Malaysian Women of Reproductive Age

ObjectivesRiboflavin and choline are essential nutrients that are metabolically inter-related and play major roles in one-carbon metabolism. Betaine can donate a methyl group to homocysteine, forming dimethylglycine (DMG) and methionine in the liver and kidney. Betaine is synthesized from choline in two irreversible reactions, the first of which is catalyzed by flavin adenine dinucleotide (FAD), a cofactor formed from riboflavin. In animal models, riboflavin status has been shown to influence choline and its metabolites betaine and DMG but there are no human studies. Here we examine whether riboflavin status modifies plasma choline, betaine, and DMG concentrations in healthy women (19–45 y).MethodsFasting blood was collected from 206 Canadian and 210 Malaysian women between 2015 and 2016. Riboflavin status was assessed using a functional biomarker, erythrocyte glutathione reductase activity coefficient (EGRac). Plasma choline, betaine, and DMG were determined by LCLC MS/MS. Plasma folate and vitamin B12 were determined using the microbiological method and an immune assay, respectively. General linear models were used to assess the independent relationship between EGRac and each of the choline metabolites with adjustment for potential confounders.Results71% of Malaysian women had EGRac ≥1.40 compared to 40% of Canadian women. Betaine, DMG, and vitamin B12 concentrations were significantly higher among Malaysian women compared to Canadian women (40.6 vs. 37.0 mmol/L, 2.7 vs. 2.4 mmol/L and 360 vs. 307 pmol/L, respectively). There were no significant associations between EGRac or riboflavin deficiency (defined as EGRac ≥1.40) and choline or its’ metabolites after adjustments for age, ethnicity, body mass index, plasma folate, and vitamin B12. In the adjusted models, plasma choline was positively associated with vitamin B12 concentrations (B = 0.002, 95% CI: 0.000, 0.003) and plasma betaine was positively associated with plasma folate (B = 0.18, 95%CI: 0.07, 0.29) and vitamin B12 (B = 0.011, 95%CI: 0.002, 0.020).ConclusionsOverall, riboflavin status was not associated with choline and its metabolites in Canadian and Malaysian women.Funding SourcesDairy Farmers of Canada, Saudi Arabian Cultural Bureau in Canada, and BC Children's Hospital Research Institute.

Relationship Between PEMT Gene rs7946 Polymorphism and Nutritional Choline Status in Association With Nonalcoholic Fatty Liver Risk

ObjectivesPhosphatidylethanolamine N-methyltransferase (PEMT) rs7946 genetic polymorphism and nutritional choline status on nonalcoholic fatty liver (NAFL) risk have not been well elucidated. The aims of the study were to investigate the choline-polymorphic relationship on NAFL risk.MethodsThe study recruited 253 patients with metabolic disorders from one regional hospital in 2020. By abdominal ultrasound diagnosis, 99 patients with hepatic steatosis (HS) and 99 age- and sex-matched controls without HS were included for the study. Habitual dietary choline intakes were assessed using validated quantitative FFQ by a face-to-face interview. PEMT rs7946 (V175M, G to A substitution) polymorphism, blood choline concentrations and metabolic disorders factors were analyzed for their interaction with HS risk.ResultsGenetic distribution of PEMT rs7946 polymorphism significantly differed in controls (GG: 48%, GA, 32%, AA, 20%) and in the HS cases (GG: 69%, GA: 26%, AA: 5%) (p = 0.003). Higher values of body mass index (BMI), serum triglycerides (TG), insulin resistance (HOMO-IR), alanine aminotransferase and lower levels of HDL were associated with HS cases. Plasma free choline levels were significantly higher in HS cases (median:12.4 μM, quartile range (9.9–14.8)) than in controls (median: 10.6 μM, quartile range (9.3–12.8)) (p = 0.001). The daily dietary intake of total choline and the choline derivatives did not differ between the two groups. After adjustment of age, gender, BMI, energy consumption, fiber intake, and blood metabolic markers (TG, LDL, HDL and HOMA-IR), odds ratios of HS were 7-fold increase (OR: 7.6, 95% CI: 2.3–24.4) for patients with PEMT rs7946 GG genotype of high plasma choline level (> median levels: 11.6 μM) as compared with GA/AA genotype of plasma choline level lower than 11.6 μM.ConclusionsThe PEMT rs7946 GG genotype in combination with high plasma choline levels was associated with increased risks of hepatic fat accumulation in patients with metabolic disorders.Funding SourcesThe Ministry of Health and Welfare-affiliated Taipei Hospital. The Ministration of Science and Technology, Taiwan, ROC.

Plasma Choline Concentration Was Not Increased After a 6-Month Egg Intervention in 6–9-Month-Old Malawian Children: Results from a Randomized Controlled Trial

ABSTRACTBackgroundEggs are a rich source of choline, an essential nutrient important for child growth and development. In a randomized trial of 1 egg/d in young children in Ecuador, an egg intervention led to significant improvements in growth, which were partially mediated by increased plasma choline concentration. A similar trial in Malawi (clinicaltrials.gov: NCT03385252) found little improvement in child growth or development.ObjectivesWe aimed to evaluate the effect of 1 egg/d for 6 mo on plasma choline concentrations in Malawian children enrolled in a randomized trial.MethodsInfants aged 6–9 mo in rural Malawi were randomly assigned to receive 1 egg/d (n = 331) or serve as a nonintervention control (n = 329) for 6 mo. Anthropometric, developmental, and dietary data were collected at baseline and 6-mo follow-up, along with a blood draw. Plasma choline, betaine, dimethylglycine, trimethylamine N-oxide (TMAO), and DHA were measured at both time points using ultrahigh performance liquid chromatography–tandem MS (n = 200 per group). Linear regression analysis was used to determine the difference in plasma choline and related metabolites between groups after 6 mo of intervention.ResultsPlasma choline, betaine, dimethylglycine, and DHA concentrations did not differ between groups at 6-mo follow-up. Plasma TMAO was significantly (26%; 95% CI: 7%, 48%) higher in the egg intervention group in a fully adjusted model.ConclusionsProvision of 1 egg/d for 6 mo did not result in increases in plasma choline or related metabolites, except TMAO. This could partially explain the lack of effect on growth and development. Additional interventions are needed to improve choline status, growth, and development in this population.

Two methods for assessment of choline status in a randomized crossover study with varying dietary choline intake in people: isotope dilution MS of plasma and in vivo single-voxel magnetic resonance spectroscopy of liver

Choline deficiency has numerous negative health consequences; although the preponderance of the US population consumes less than the recommended Adequate Intake (AI), clinical assessment of choline status is difficult. Further, several pathways involved in primary metabolism of choline are estrogen-sensitive and the AI for premenopausal women is lower than that for men. We sought to determine whether in vivo magnetic resonance spectroscopy (MRS) of liver and/or isotope-dilution MS of plasma could identify biomarkers reflective of choline intake (preregistered primary outcomes 1 and 2, secondary outcome 1). Determination of whether biomarker concentrations showed sex dependence was a post hoc outcome. This substudy is a component of a larger project to identify a clinically useful biomarker panel for assessment of choline status. In a double-blind, randomized, crossover trial, people consumed 3 diets, representative of ∼100%, ∼50%, and ∼25% of the choline AI, for 2-wk periods. We measured the concentrations of choline and several metabolites using 1H single-voxel MRS of liver in vivo and using 2H-labeled isotope dilution MS of several choline metabolites in extracted plasma. Plasma concentrations of 2H9-choline, unlabeled betaine, and 2H9-betaine, and the isotopic enrichment ratio (IER) of betaine showed highly significant between-diet effects (q<0.0001), with unlabeled betaine concentration decreasing 32% from highest to lowest choline intake. Phosphatidylcholine IER was marginally significant (q=0.03). Unlabeled phosphatidylcholine plasma concentrations did not show between-diet effects (q=0.34). 2H9 (trimethyl)-phosphatidylcholine plasma concentrations (q=0.07) and MRS-measured total soluble choline species liver concentrations (q=0.07) showed evidence of between-diet effects but this was not statistically significant. Although MRS is a more direct measure of choline status, variable spectral quality limited interpretation. MS analysis of plasma showed clear correlation of plasma betaine concentration, but not plasma phosphatidylcholine concentration, with dietary choline intake. Plasma betaine concentrations also correlate with sex status (premenopausal women, postmenopausal women, men).This trial was registered at clinicaltrials.gov as NCT03726671.

Effects of Plasma Choline Concentrations on Placental Development and Fetal Growth, With Potential Mechanistic Roles of Imprinted Genes

ObjectivesImprinted genes are epigenetically regulated and play critical roles in placental development and fetal growth. We aimed to examine (1) the impact of maternal one-carbon (methyl donor) nutrition on placental imprinted gene expression, placental development, and fetal growth; (2) whether imprinted gene expression alterations mediate effects of one-carbon nutrition on placental development and fetal growth; (3) interaction effects between one-carbon nutrients and imprinted genes in placental development and fetal growth. MethodsHistopathology and expression of 109 imprinted genes (Nanostring) were assessed in placentas from 101 women recruited at initiation of antenatal care in a prospective cohort study examining developmental effects of prenatal alcohol exposure in South Africa. Women were interviewed prenatally about demographics, alcohol, smoking, and drug use, and erythrocyte folate, serum vitamin B12, and plasma choline concentrations were assayed at recruitment. Infant weight and height were assessed at age 2 wk. ResultsIn limma tests, women with plasma choline concentrations below the median had lower placental expression of EPS15, IGF2R, LINC00657, SGCE, ZC3H12C, and ZNF264 than women above the median (p < .05, FDR < .10). In regression models adjusted for potential confounders (maternal age, gravidity, education, alcohol and drug use), plasma choline (μM) was associated with larger placental weight (g) (B = 14.0(1.9, 26.2)) and reduced maternal vascular underperfusion (MVU) prevalence (B = –.07(–.12, –.02). In causal inference analyses, there were trends for mediation of the relation between choline and MVU by decreased LINC00657, ZC3H12C, and ZNF264 expression. In regression models examining plasma choline X imprinted gene expression interaction effects, choline modified relations of EPS15, ZC3H12C, and ZNF264 to placental weight and fetal growth. ConclusionsMaternal plasma choline was associated with decreased placental expression of 6 imprinted genes, 3 of which may mediate effects of choline on placental development. Choline modified effects of 3 genes on placental and fetal growth. These findings suggest maternal choline status may impact placental and fetal development, with imprinted genes playing mechanistic roles. Funding SourcesNIH/NIAAA; Lycaki-Young Fund.

High-Throughput Analysis of Water-Soluble Forms of Choline and Related Metabolites in Human Milk by UPLC-MS/MS and Its Application.

Choline and related metabolites are key factors in many metabolic processes, and insufficient supply can adversely affect reproduction and fetal development. Choline status is mainly regulated by intake, and human milk is the only choline source for exclusively breastfed infants. Further, maternal status, genotype, and phenotype, as well as infant outcomes, have been related to milk choline concentrations. In order to enable the rapid assessment of choline intake for exclusively breastfed infants and to further investigate the associations between milk choline and maternal and infant status and other outcomes, we have developed a simplified method for the simultaneous analysis of human milk choline, glycerophosphocholine, phosphocholine, and the less abundant related metabolites betaine, carnitine, creatinine, dimethylglycine (DMG), methionine, and trimethylamine N-oxide (TMAO) using ultraperformance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). These analytes have milk concentrations ranging over 3 orders of magnitude. Unlike other recently described LC-based methods, our approach does not require an ion-pairing reagent or high concentrations of solvent modifiers for successful analyte separation and thus avoid signal loss and potential permanent contamination. Milk samples (10 μl) were diluted (1:80) in water : methanol (1:4, v:v) and filtered prior to analysis with an optimized gradient of 0.1% propionic acidaq and acetonitrile, allowing efficient separation and removal of contaminants. Recovery rates ranged from 108.0 to 130.9% (inter-day variation: 3.3–9.6%), and matrix effects (MEs) from 54.1 to 114.3%. MEs were greater for carnitine, creatinine, and TMAO at lower dilution (1:40, p < 0.035 for all), indicating concentration-dependent ion suppression. Milk from Brazilian women (2–8, 28–50, and 88–119 days postpartum, ntotal = 53) revealed increasing concentration throughout lactation for glycerophosphocholine, DMG, and methionine, while carnitine decreased. Choline and phosphocholine were negatively correlated consistently at all three collection time intervals. The method is suitable for rapid analysis of human milk water-soluble forms of choline as well as previously not captured related metabolites with minimal sample volumes and preparation.

Two methods for assessment of choline status in a randomized crossover study with varying dietary choline intake in people: Isotope dilution MS of plasma and in vivo single-voxel magnetic resonance spectroscopy of liver

Two methods for assessment of choline status in a randomized crossover study with varying dietary choline intake in people: Isotope dilution MS of plasma and in vivo single-voxel magnetic resonance spectroscopy of liver

Effects of maternal and fetal choline concentrations on the fetal growth and placental DNA methylation of 12 target genes related to fetal growth, adipogenesis, and energy metabolism.

We performed a birth cohort study involving 124 mother-infant pairs to investigate whether placental DNA methylation is associated with maternal choline status and fetal development. Plasma choline concentration was assayed longitudinally in the 1st and 3rd trimesters and at term-pregnancy in mothers and cord blood. Placental DNA methylation was measured for 12 target candidate genes that are related to fetal growth, adipogenesis, lipid and energy metabolism, or long interspersed nuclear elements. Higher maternal plasma and cord blood choline levels at term tended to associate with lower birthweight (r = -0.246, P < 0.013; r = -0.290, P < 0.002) and body mass index (BMI) at birth (r = 0.344, P < 1E-3; r = -0.360, P < 1E-3). The correlation between maternal plasma choline level and cord blood choline level was relatively modest (r = 0.049, P = 0.639). There was an inverse correlation between placental DNA methylation at the retinoid X receptor alpha (RXRA) gene and maternal plasma choline level (r = -0.188 to r = -0.452, P = 0.043 to P < 1E-3 at three points). RXRA methylation level was positively associated with birthweight and BMI at birth (r = 0.306, P = 0.001; r = 0.390, P < 1E-3). Further, RXRA methylation was inversely correlated with RXRA gene expression level (r = 0.333, P < 1E-3). Our results suggest that the association between maternal choline status and placental RXRA methylation represents a potential fetal programing mechanism contributing to fetal growth.

Nutrition and neurodevelopment: the search for candidate nutrients in the Seychelles Child Development Nutrition Study

This review examines the role of nutrients in child development and outlines the key nutrients identified as potentially important to neurodevelopment among high fish consumers in the Seychelles Child Development Nutrition Study (SCDNS). It describes the clinical assessment of these nutrients in the blood and breast milk samples collected from the cohort of 300 pregnant women who were recruited, at their first antenatal visit, on the SCDNS. These key nutrients include the long chain polyunsaturated fatty acids (LCPUFA), docosohexaenoic acid (DHA) and arachidonic acid (AA), both of which may affect neurodevelopment in the later stages of fetal growth. Only DHA, however, is strongly associated with fish consumption, the predominant source of the neurotoxicant methyl mercury (MeHg). Any benefits of increased selenium status on neurodevelopment are likely to accrue via detoxification of MeHg during fetal growth, while benefits of optimal iodine or thyroid status are likely to be directly related to neurodevelopment during late fetal growth. Unlike LCPUFA, Se, and I, the status of the B vitamins, folate, vitamin B12, vitamin B6, and riboflavin are unlikely to be closely related to fish consumption but the status of each of these B vitamins is likely to impinge on overall status of choline, which is expected to have direct effects on neurodevelopment both prenatally and postnatally and may also impact on MeHg toxicity. Choline status, together with the status of two other candidate nutrients, zinc and copper, which are also likely to have effects on neurodevelopment prenatally and postnatally, are expected to have some correlation with fish consumption.

Betaine and choline status modify the effects of folic acid and creatine supplementation on arsenic methylation in a randomized controlled trial of Bangladeshi adults.

Methylation of ingested inorganic arsenic (InAs) to monomethyl- (MMAs) and dimethyl-arsenical species (DMAs) facilitates urinary arsenic elimination. Folate and creatine supplementation influenced arsenic methylation in a randomized controlled trial. Here, we examine if baseline status of one-carbon metabolism nutrients (folate, choline, betaine, and vitamin B12) modified the effects of FA and creatine supplementation on changes in homocysteine, guanidinoacetate (GAA), total blood arsenic, and urinary arsenic metabolite proportions and indices. Study participants (N = 622) received 400 or 800μg FA, 3g creatine, 400μg FA + 3g creatine, or placebo daily for 12weeks. Relative to placebo, FA supplementation was associated with greater mean increases in %DMAs among participants with betaine concentrations below the median than those with levels above the median (FDR < 0.05). 400μg FA/day was associated with a greater decrease in homocysteine among participants with plasma folate concentrations below, compared with those above, the median (FDR < 0.03). Creatine treatment was associated with a significant decrease in %MMAs among participants with choline concentrations below the median (P = 0.04), but not among participants above the median (P = 0.94); this effect did not significantly differ between strata (P = 0.10). Effects of FA and creatine supplementation on arsenic methylation capacity were greater among individuals with low betaine and choline status, respectively. The efficacy of FA and creatine interventions to facilitate arsenic methylation may be modified by choline and betaine nutritional status. Clinical Trial Registry Identifier: NCT01050556, U.S. National Library of Medicine, https://clinicaltrials.gov ; registered January 15, 2010.

Plasma Free Choline Concentration Did Not Reflect Dietary Choline Intake in Early and Late Pregnancy: Findings from the APrON Study

Abstract Objectives Choline is a critical nutrient for fetal development. Pregnancy studies showed that most women have choline intakes below the adequate intake (AI) level of 450 mg/d. Research on plasma free choline as an indicator of dietary choline intake showed conflicting results to date. We sought to compare plasma free choline concentration between women with different choline intakes and to explore the association between plasma free choline and dietary choline intake in early (EP) and late pregnancy (LP). Methods This study included data and non-fasting plasma samples from pregnant women enrolled in the Alberta Pregnancy Outcomes and Nutrition (APrON) cohort study. EP (&amp;lt;20 weeks of gestation) and LP (&amp;gt;20 weeks of gestation) dietary choline intake was estimated using a 24-hr recall. Two categories of dietary choline intake were created: 1) low choline (LCI), i.e., choline intake in 1st quartile (Q) in EP, with these women having choline intake in 1st or 2nd Q in LP (n = 61); 2) high choline intake (HCI), i.e., choline intake in 4th Q in EP and in 3rd or 4th Q in LP (n = 46). Linear mixed-effects models were used to explore the association between plasma free choline and dietary choline intake across EP and LP, after adjustment for maternal age, ethnicity and weeks of gestation. Results Median (IQR) maternal age was 32 (30–35) y, and 80% were Caucasian. LCI was 101 (86–109) and 109 (93–127) mg/day in EP and LP, respectively, and HCI was 251 (223–286) and 212 (177–274) mg/day. Plasma free choline (μmol/L) did not differ between LCI and HCI at EP [LCI: 10.6 (9.03, 12.9); HCI: 11.7 (10.2, 13.8)] and LP [LCI: 11.7 (10.6, 12.7); HCI: 12.7 (10.7, 15.8)] (P &amp;gt; 0.05, Wilcoxon rank-sum test). Per 10 mg of choline intake, plasma free choline increased by 0.34 (95%CI 0.12, 0.56) in those with LCI, and 0.18 (95%CI 0.050, 0.31) in women with HCI, across EP and LP after adjustment. Conclusions In this subgroup of pregnant women, plasma free choline concentration did not reflect differences in dietary choline intake in EP or LP. This may be explained by an overall low choline intake (&amp;lt;AI) which would promote rapid tissue uptake of choline. The identification of a sensitive and dynamic biomarker for choline status is required. Funding Sources UBC Four Year Doctoral Fellowship, Canada Research Chair Program, CIHR NTE Grant FRN 160,942, Alberta Innovates for the APrON cohort.

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.

Effect of Maternal Nonalcoholic Fatty Liver Disease and Dietary Choline Status on Body Mass and Lipid Profile in Rat Offspring

AbstractBoth maternal metabolic status and nutrition during pregnancy and lactation may have a programming effect on offspring metabolism. The aim of this study was to examine the role of the dietary choline supply during pregnancy and lactation in rat dams suffering from nonalcoholic fatty liver disease (NAFLD) on body weight and plasma lipid profile of the progeny.The research protocol was approved by the local ethics committee. The study groups included the offspring of 1. healthy dams receiving choline during pregnancy and lactation (the control group); 2. NAFLD dams receiving choline during pregnancy and lactation (NN); 3. NAFLD dams receiving choline during pregnancy and a choline-deficient diet during lactation (ND); 4. NAFLD dams receiving a choline-deficient diet during pregnancy and a supply of choline during lactation (DN); and 5. NAFLD dams receiving a choline-deficient diet during both pregnancy and lactation (DD). Body mass and plasma lipid profile were assessed in male and female rats from each group on day 3 (3d), day 24 (24d), and day 90 (90d).Body mass was significantly lower in the male offspring of the DD and DN groups than in the control group. Differences were observed at all times (3d: p = 0.0023; 24d: p &lt; 0.0001; 90d: p &lt; 0.0001). Moreover, body mass was significantly higher in the male offspring of the control group than in any other group. In the female progeny, body mass was higher in the control group than in the ND (24d: p &lt; 0.0001; 90d: p = 0.0067) or NN (24d: p = 0.0058) groups.Total plasma cholesterol concentration was higher in the 90d males of the control group than in the DD group (p = 0.0163) and in the 24d females of the NN group than in the ND group (p = 0.0495). In the 3d animals, LDL was higher (p = 0.0083) but HDL was lower (p = 0.0196) in male rats of the DD and DN groups than in the NN and ND groups. Neither age nor sex affected LDL levels. The plasma levels of triglycerides were not affected by the dietary regimen, sex, or age of the animals.Maternal NAFLD and dietary choline status during pregnancy and lactation affect body mass and lipid profile in rat offspring, and the effects of maternal programming are more pronounced in male offspring than in female.The project was financed by the National Science Centre, Poland (2016/21/D/NZ9/00360).