PSVI-29 Late-Breaking: Exposure to 1.8 mM choline chloride does not impact development to the blastocyst stage or pregnancy rate after transfer for embryos produced in vitro

Choline is a nutrient that plays a role as a precursor for the neurotransmitter acetylcholine, the membrane phospholipid phosphatidylcholine, and the methyl donor betaine. Embryos produced in vitro are usually cultured without an exogenous choline source. We hypothesized that exposure to 1.8 mM choline chloride would increase percent of embryos becoming blastocysts in culture and pregnancy rate after transfer of embryos into recipients. A total of 39 Brahman and Senepol donors were used to produce embryos for transfer into recipient crossbred females. Donors were assigned to have their embryos cultured in either 1.8 mM choline chloride or, as a control, 1.8 mM extra NaCl. The percent of oocytes cleaved were measured 3 days after insemination and percent blastocyst at day 7.5. Embryos were transferred into recipient cows and pregnancy was diagnosed at 28–31 days of gestation and then confirmed at 50–56 days. Data were analyzed using PROC GLIMMIX in SAS. Treatment did not affect cleavage rate (67.3 + 1.6 vs 68.6 + 1.6% for choline vs control; P = .0.5632) or percent of cleaved embryos becoming blastocysts (17.6 + 1.3 vs 18.1 + 1.3%; P = 0.5355). Similarly, there was no effect of treatment on pregnancy days 28–31 [42.5% (48/113 cows) vs 47.3% (54/114 cows) for choline vs control; P = 0.4339] or at days 50–56 [39.1% (36/92) vs 38.5% (32/83); P = 0.5348]. In summary, 1.8 mM choline chloride does not impact embryo development to the blastocyst stage or pregnancy establishment. Further investigation is needed to evaluate the phenotype of the subsequent calves to determine whether early exposure to choline has consequences for postnatal function. Support: USDA-NIFA 2020-67015-30821.

Pharmacokinetic properties of a novel formulation of S-adenosyl-l-methionine phytate

S-adenosyl-l-methionine (SAM), the main endogenous methyl donor, is the adenosyl derivative of the amino acid methionine, which displays many important roles in cellular metabolism. It is widely used as a food supplement and in some countries is also marketed as a drug. Its interesting nutraceutical and pharmacological properties prompted us to evaluate the pharmacokinetics of a new form of SAM, the phytate salt. The product was administered orally to rats and pharmacokinetic parameters were evaluated by comparing the results with that obtained by administering the SAM tosylated form (SAM PTS). It was found that phytate anion protects SAM from degradation, probably because of steric hindrance exerted by the counterion, and that the SAM phytate displayed significant better pharmacokinetic parameters compared to SAM PTS. These results open to the perspective of the use of new salts of SAM endowed with better pharmacokinetic properties.

Fatty acid metabolism and colon cancer protection by dietary methyl donor restriction

Introduction A methyl donor depleted (MDD) diet dramatically suppresses intestinal tumor development in Apc-mutant mice, but the mechanism of this prevention is not entirely clear. Objectives We sought to gain insight into the mechanisms of cancer suppression by the MDD diet and to identify biomarkers of cancer risk reduction. Methods A plasma metabolomic analysis was performed on ApcΔ14/+ mice maintained on either a methyl donor sufficient (MDS) diet or the protective MDD diet. A group of MDS animals was also pair-fed with the MDD mice to normalize caloric intake, and another group was shifted from an MDD to MDS diet to determine the durability of the metabolic changes. Results In addition to the anticipated changes in folate one-carbon metabolites, plasma metabolites related to fatty acid metabolism were generally decreased by the MDD diet, including carnitine, acylcarnitines, and fatty acids. Some fatty acid selectivity was observed; the levels of cancer-promoting arachidonic acid and 2-hydroxyglutarate were decreased by the MDD diet, whereas eicosapentaenoic acid (EPA) levels were increased. Machine-learning elastic net analysis revealed a positive association between the fatty acid-related compounds azelate and 7-hydroxycholesterol and tumor development, and a negative correlation with succinate and β-sitosterol. Conclusion Methyl donor restriction causes dramatic changes in systemic fatty acid metabolism. Regulating fatty acid metabolism through methyl donor restriction favorably effects fatty acid profiles to achieve cancer protection.

Dietary methyl donor micronutrients intake in relation to psychological disorders in adults

Previous investigations have mostly studied an individual methyl donor nutrient in relation to psychological disorders and the findings were inconsistent. We investigated the association of methyl donor micronutrients (folate, B6, B12, choline, betaine, and methionine) with psychological disorders (depression, anxiety, psychological distress) in Iranian adults. In this cross-sectional study, dietary intakes of 3299 adults were collected using a validated food frequency questionnaire. Methyl donor micronutrient score (MDMS) was calculated based on energy-adjusted deciles of each nutrient. Hospital Anxiety and Depression Scale (HADS) and General Health Questionnaire (GHQ), validated for Iranians, have been applied to assess depression, anxiety, and psychological distress. Participants had a mean age of 36.3±7.9 years, of whom 58.5% were women. After considering potential confounders, adults in the top quartile of MDMS, compared to the bottom one, had decreased odds of anxiety (OR: 0.53, 95%CI: 0.37–0.75), depression (OR: 0.75, 95%CI: 0.58–0.97) and psychological distress (OR: 0.61, 95%CI: 0.46–0.80). Stratified analysis revealed that the highest category of MDMS among men was related to a 68% lower odds of anxiety (95%CI: 0.15–0.68). Among women, the top quartile of MDMS was protectively associated with anxiety (OR: 0.60, 95%CI: 0.40–0.90), depression (OR: 0.68, 95%CI: 0.50–0.93) and psychological distress (OR: 0.53, 95%CI: 0.38–0.74). Overweight and obese subjects in the highest quartile of MDMS had a 67%, 35%, and 53% lower odds of anxiety (95%CI: 0.20–0.56), depression (95%CI: 0.44–0.94), and psychological distress (95%CI: 0.31–0.70), respectively. We found that high consumption of methyl donor micronutrients was related to a reduced odds of psychological disorders, especially in women and overweight or obese individuals. Further prospective studies are needed to affirm these findings.

Methyl donor micronutrients, CD40-ligand methylation and disease activity in systemic lupus erythematosus: A cross-sectional association study

Objective Hypomethylation of CD40-ligand (CD40L) in T-cells is associated with increased disease activity in systemic lupus erythematosus (SLE). We therefore investigated possible associations of dietary methyl donors and products with CD40L methylation status in SLE. Methods Food frequency questionnaires were employed to calculate methyl donor micronutrients in 61 female SLE patients (age 45.7 ± 12.0 years, disease duration 16.2 ± 8.4 years) and compared to methylation levels of previously identified key DNA methylation sites (CpG17 and CpG22) within CD40L promotor of T-cells using quantitative DNA methylation analysis on the EpiTYPER mass spectrometry platform. Disease activity was assessed by SLE Disease Activity Index (SLEDAI). Linear regression modelling was used. P values were adjusted according to Benjamini & Hochberg. Results Amongst the micronutrients assessed (g per day), methionine and cysteine were associated with methylation of CpG17 (β = 5.0 (95%CI: 0.6-9.4), p = 0.04; and β = 2.4 (0.6-4.1), p = 0.02, respectively). Methionine, choline, and cysteine were additionally associated with the mean methylation of the entire CD40L (β = 9.5 (1.0-18.0), p = 0.04; β = 1.6 (0.4-3.0), p = 0.04; and β = 4.3 (0.9-7.7), p = 0.02, respectively). Associations of the SLEDAI with hypomethylation were confirmed for CpG17 (β=-32.6 (-60.6 to -4.6), p = 0.04) and CpG22 (β=-38.3 (-61.2 to -15.4), p = 0.004), but not the mean methylation of CD40L. Dietary products with the highest impact on methylation included meat, ice cream, white bread, and cooked potatoes. Conclusions Dietary methyl donors may influence DNA methylation levels and thereby disease activity in SLE.

Methyl-Donor Micronutrient for Gestating Sows: Effects on Gut Microbiota and Metabolome in Offspring Piglets

This study aimed to investigate the effects of maternal methyl-donor micronutrient supplementation during gestation on gut microbiota and the fecal metabolic profile in offspring piglets. Forty-three Duroc × Erhualian gilts were assigned to two dietary groups during gestation: control diet (CON) and CON diet supplemented with MET (folic acid, methionine, choline, vitamin B6, and vitamin B12). The body weights of offspring piglets were recorded at birth and weaning. Besides this, fresh fecal samples of offspring piglets were collected at 7, 14, and 21 days. The gut microbiota composition, metabolic profile, and short-chain fatty acid (SCFA) profiles in the fecal samples were determined using 16S rDNA sequencing, liquid chromatography-mass spectrometry metabolomics, and gas chromatography methods, respectively. The results showed that maternal methyl-donor micronutrient supplementation increased the microbiota diversity and uniformity in feces of offspring piglets as indicated by increased Shannon and Simpson indices at 7 days, and greater Simpson, ACE, Chao1 and observed species indices at 21 days. Specifically, at the phylum level, the relative abundance of Firmicutes and the Firmicutes to Bacteroidetes ratio were elevated by maternal treatment. At the genus level, the relative abundance of SCFA-producing Dialister, Megasphaera, and Turicibacter, and lactate-producing Sharpea as well as Akkermansia, Weissella, and Pediococcus were increased in the MET group. The metabolic analyses show that maternal methyl-donor micronutrient addition increased the concentrations of individual and total SCFAs of 21-day piglets and increased metabolism mainly involving amino acids, pyrimidine, and purine biosynthesis. Collectively, maternal methyl-donor micronutrient addition altered gut microbiota and the fecal metabolic profile, resulting in an improved weaning weight of offspring piglets.

The BHMT-betaine methylation pathway epigenetically modulates oligodendrocyte maturation

Research into the epigenome is of growing importance as a loss of epigenetic control has been implicated in the development of neurodegenerative diseases. Previous studies have implicated aberrant DNA and histone methylation in multiple sclerosis (MS) disease pathogenesis. We have previously reported that the methyl donor betaine is depleted in MS and is linked to changes in histone H3 trimethylation (H3K4me3) in neurons. We have also shown that betaine increases histone methyltransferase activity by activating chromatin bound betaine homocysteine S-methyltransferase (BHMT). Here, we investigated the role of the BHMT-betaine methylation pathway in oligodendrocytes. Immunocytochemistry in the human MO3.13 cell line, primary rat oligodendrocytes, and tissue from MS postmortem brain confirmed the presence of the BHMT enzyme in the nucleus in oligodendrocytes. BHMT expression is increased 2-fold following oxidative insult, and qRT-PCR demonstrated that betaine can promote an increase in expression of oligodendrocyte maturation genes SOX10 and NKX-2.2 under oxidative conditions. Chromatin fractionation provided evidence of a direct interaction of BHMT on chromatin and co-IP analysis indicates an interaction between BHMT and DNMT3a. Our data show that both histone and DNA methyltransferase activity are increased following betaine administration. Betaine effects were shown to be dependent on BHMT expression following siRNA knockdown of BHMT. This is the first report of BHMT expression in oligodendrocytes and suggests that betaine acts through BHMT to modulate histone and DNA methyltransferase activity on chromatin. These data suggest that methyl donor availability can impact epigenetic changes and maturation in oligodendrocytes.