Effect of long-term betaine supplementation on chemical and physical characteristics of three muscles from the Alentejano pig

BACKGROUND: The use of betaine as a dietary supplement to reduce fat deposition and increase lean muscle mass in pigs is increasing. However, there is no available information on its effect on the physical and chemical (i.e. fatty acid (FA) profile) characteristics of pork. RESULTS: The effects of long-term betaine intake (1 g betaine kg(-1) diet for 20 weeks) on the chemical and physical characteristics of Musculus longissimus dorsi, M. biceps femoris and M. semimembranosus from the Alentejano pig were investigated. The contents of total protein, intramuscular lipids (neutral and polar), myoglobin and total collagen as well as the water-holding capacity, physical colour characteristics and FA profile of the three muscles were determined. With the exception of a significant increase in the intramuscular lipid content of M. longissimus dorsi and M. biceps femoris, betaine supplementation did not influence muscle chemical and colour characteristics or body fat deposition. CONCLUSION: These data suggest that long-term betaine supplementation selectively increases intramuscular lipid deposition (a trait related to meat quality) while not affecting other chemical (including the atheroprotective FA profile) and physical characteristics of the Alentejano pig muscles studied.
Martins, J.M., et al., Effect of long-term betaine supplementation on chemical and physical characteristics of three muscles from the Alentejano pig. J Sci Food Agric, 2012. 92(10): p. 2122-7

Plasma free choline, betaine and cognitive performance: the Hordaland Health Study

Choline and betaine are nutrients involved in one-carbon metabolism. Choline is essential for neurodevelopment and brain function. We studied the associations between cognitive function and plasma concentrations of free choline and betaine. In a cross-sectional study, 2195 subjects (55 % women), aged 70-74 years, underwent extensive cognitive testing including the Kendrick Object Learning Test (KOLT), Trail Making Test (part A, TMT-A), modified versions of the Digit Symbol Test (m-DST), Block Design (m-BD), Mini-Mental State Examination (m-MMSE) and Controlled Oral Word Association Test (COWAT). Compared with low concentrations, high choline (>8.4 mumol/l) was associated with better test scores in the TMT-A (56.0 v. 61.5, P = 0.004), m-DST (10.5 v. 9.8, P = 0.005) and m-MMSE (11.5 v. 11.4, P = 0.01). A generalised additive regression model showed a positive dose-response relationship between the m-MMSE and choline (P = 0.012 from a corresponding linear regression model). Betaine was associated with the KOLT, TMT-A and COWAT, but after adjustments for potential confounders, the associations lost significance. Risk ratios (RR) for poor test performance roughly tripled when low choline was combined with either low plasma vitamin B12 ( </= 257 pmol/l) concentrations (RRKOLT = 2.6, 95 % CI 1.1, 6.1; RRm-MMSE = 2.7, 95 % CI 1.1, 6.6; RRCOWAT = 3.1, 95 % CI 1.4, 7.2) or high methylmalonic acid (MMA) ( >/= 3.95 mumol/l) concentrations (RRm-BD = 2.8, 95 % CI 1.3, 6.1). Low betaine ( </= 31.1 mumol/l) combined with high MMA was associated with elevated RR on KOLT (RRKOLT = 2.5, 95 % CI 1.0, 6.2). Low plasma free choline concentrations are associated with poor cognitive performance. There were significant interactions between low choline or betaine and low vitamin B12 or high MMA on cognitive performance.

Nurk, E., et al., Plasma free choline, betaine and cognitive performance: the Hordaland Health Study. Br J Nutr, 2012 (May 1): p. 1-9

Deletion of murine betaine-homocysteine S-methyltransferase in mice perturbs choline and 1-carbon metabolism, resulting in fatty liver and hepatocellular carcinoma

Betaine-homocysteine S-methyltransferase (BHMT) uses betaine to catalyze the degradation of homocysteine (Hcy). There are common genetic polymorphisms in the BHMT gene in humans. To model the phenotype of mice with a loss of BHMT function, we generated the first Bhmt-/- mouse. Deletion of the gene resulted in a 6-fold increase in hepatic and an 8-fold increase in plasma Hcy concentrations, suggesting the importance of BHMT in Hcy removal. Deletion of the gene resulted in a 43% reduction in hepatic S-adenosylmethionine (AdoMet) and a 3-fold increase in hepatic S-adenosylhomocysteine (AdoHcy) concentrations, resulting in a 75% reduction in methylation potential (AdoMet:AdoHcy). Bhmt-/- mice accumulated betaine in most tissues, including a 21-fold increase in the liver concentration compared to wildtype (WT). These mice had lower concentrations of choline, phosphocholine, glycerophosphocholine, phosphatidylcholine and sphingomyelin in several tissues. At 5 weeks of age, Bhmt-/- mice had 36% lower total hepatic phospholipid concentrations and a 6-fold increase in hepatic triacyglycerol concentrations compared to WT, which was due to a decrease in the secretion of very low density lipoproteins. At 1 year of age, 64% of Bhmt-/- mice had visible hepatic tumors. Histopathological analysis revealed that Bhmt-/- mice developed hepatocellular carcinoma (HCC) or carcinoma precursors. These results indicate that BHMT has an important role in Hcy, choline and one-carbon homeostasis. A lack of BHMT also affects susceptibility to fatty liver and HCC. We suggest that functional polymorphisms in BHMT that significantly reduce activity may have similar effects in humans.

Teng, Y.W., et al., Deletion of murine betaine-homocysteine S-methyltransferase in mice perturbs choline and 1-carbon metabolism, resulting in fatty liver and hepatocellular carcinoma. J Biol Chem, 2011. 286: p. 36258-67

Thermal and refining processes, not fermentation, tend to reduce lipotropic capacity of plant-based foods

Plant-based foods (PBF) are relevant and diversified sources of lipotropes, which are compounds preventing excess hepatic fat deposits. In a first study, we defined the lipotropic capacity (LC, %) of raw PBF as the means of 8 lipotrope densities (LD, mg/100 kcal), each expressed relative to that of a reference food ranking the highest considering its mean 8 LD ranks (LCraw asparagus = 100%) (A. Fardet, J.-F. Martin and J. M. Chardigny, J. Food Comp. Anal., 2011, DOI: 10.1016/j.jfca.2011.1003.1013). We showed that vegetables appeared as the best source of lipotropes on a 100 kcal-basis compared to legumes, cereals, fruits and nuts. The main objective of this second study was to quantify the effect of processing on LD and LC of raw PBF based on lipotrope contents collected in a USDA (United State Department of Agriculture) database and the literature, i.e. betaine, choline, myo-inositol, methionine, magnesium, niacin, pantothenic acid and folate contents. Choline and betaine densities were not significantly affected by processing while methionine and lipotropic micronutrient densities were significantly decreased, especially for magnesium, pantothenate and folates. Myo-inositol density decreases were insignificant due to lower product number resulting from limited literature data. Lipotropic micronutrient densities were more affected by processing than other densities. Fermentations increased betaine (median change of +32%) and choline (+34%) densities. Canning and boiling vegetables increased choline densities (+26%). Globally, processing significantly reduced LC by 20%, fermentations being less drastic (median change of −5%) than refining (−33%) and thermal treatments (−16%). More specifically, canning increased LC of beetroot (536 vs 390%) and common bean (40 vs 36%) as fermentation towards LC grape (14 vs 7% for wine). Results were then mainly discussed based on percentages of lipotrope content changes on a dry-weight basis. Results of this study also showed that the LC is quite a relevant index to estimate effect of processing on lipotropic potential of PBF.

Fardet, A., J.F. Martin, and J.M. Chardigny, Thermal and refining processes, not fermentation, tend to reduce lipotropic capacity of plant-based foods. Food Funct, 2011. 2(8): p. 483-504.

The regulation of non-coding RNA expression in the liver of mice fed DDC

Mallory–Denk bodies (MDBs) are found in the liver of patients with alcoholic and chronic nonalcoholic liver disease, and hepatocellular carcinoma (HCC). Diethyl 1,4-dihydro-2,4,6,-trimethyl-3,5-pyridinedicarboxylate (DDC) is used as a model to induce the formation of MDBs in mouse liver. Previous studies in this laboratory showed that DDC induced epigenetic modifications in DNA and histones. The combination of these modifications changes the phenotype of the MDB forming hepatocytes, as indicated by the marker FAT10. These epigenetic modifications are partially prevented by adding to the diet S-adenosylmethionine (SAMe) or betaine, both methyl donors. The expression of three imprinted ncRNA genes was found to change in MDB forming hepatocytes, which is the subject of this report. NcRNA expression was quantitated by real-time PCR and RNA FISH in liver sections. Microarray analysis showed that the expression of three ncRNAs was regulated by DDC: up regulation of H19, antisense Igf2r (AIR), and down regulation of GTL2 (also called MEG3). S-adenosylmethionine (SAMe) feeding prevented these changes. Betaine, another methyl group donor, prevented only H19 and AIR up regulation induced by DDC, on microarrays. The results of the SAMe and betaine groups were confirmed by real-time PCR, except for AIR expression. After 1 month of drug withdrawal, the expression of the three ncRNAs tended toward control levels of expression. Liver tumors that developed also showed up regulation of H19 and AIR. The RNA FISH approach showed that the MDB forming cells' phenotype changed the level of expression of AIR, H19 and GTL2, compared to the surrounding cells. Furthermore, over expression of H19 and AIR was demonstrated in tumors formed in mice withdrawn for 9 months. The dysregulation of ncRNA in MDB forming liver cells has been observed for the first time in drug-primed mice associated with liver preneoplastic foci and tumors.

Oliva, J., et al., The regulation of non-coding RNA expression in the liver of mice fed DDC. Experimental and Molecular Pathology, 2009. 87(1): p. 12-19.

Maternal intake of methyl-donor nutrients and child cognition

Methyl-donor nutrients are substrates for methylation reactions involved in neurodevelopment processes. The role of maternal intake of these nutrients on cognitive performance of the offspring is poorly understood. We examined the associations of maternal intake of folate, vitamin B12, choline, betaine and methionine during the first and second trimesters of pregnancy, with tests of cognitive performance in the offspring at 3 years of age using data from 1210 participants in Project Viva, a prospective pre-birth cohort study in Massachusetts. We assessed nutrient intake with the use of food frequency questionnaires. Children's cognition at age 3 years was evaluated with the Peabody Picture Vocabulary Test III (PPVT-III) and visual-motor skills with the Wide Range Assessment of Visual Motor Abilities test. In multivariable models adjusting for potential sociobehavioural and nutritional confounders, for each 600 microg/day increment in total folate intake during the first trimester, PPVT-III score at age 3 years was 1.6 points [95% confidence interval (CI) 0.1, 3.1; P = 0.04] higher. There was a weak inverse association between vitamin B12 intake during the second trimester and PPVT-III scores [-0.4 points per 2.6 microg/day; 95% CI -0.8, -0.1; P = 0.01]. We did not find associations between choline, betaine or methionine and cognitive outcomes at this age. Results of this study suggest that higher intake of folate in early pregnancy is associated with higher scores on the PPVT-III, a test of receptive language that predicts overall intelligence, at age 3 years.

In unadjusted analyses, intake of folate and betaine during the first or second trimester was directly related to PPVT-III scores. After adjustment for potential socio-economic, educational and dietary confounders, however, the associations with both nutrients were attenuated and only the relationship between folate intake during the first trimester and PPVT-III scores remained statistically significant.

Villamor, E., et al., Maternal intake of methyl-donor nutrients and child cognition at 3 years of age. Paediatr Perinat Epidemiol, 2012. 26(4): p. 328-35.

Dietary choline and betaine intakes and risk of cardiovascular diseases: review of epidemiological evidence

BACKGROUND: Cardiovascular diseases (CVD) are the most important causes of human mortality in the world. Higher intakes of choline and betaine have been shown to be associated with lower plasma homocysteine levels (the putative CVD risk factor). This study aimed to review the evidence on the association between dietary intakes of choline and betaine and traditional/novel CVD risk factors.

METHODS: We searched in PubMed website from 1990 to 2009, with the use of following keywords: "dietary choline and betaine, cardiovascular diseases, metabolic syndrome, inflammation". The cross-sectional and prospective studies as well as the clinical trials were recruited in this investigation.

RESULTS: Dietary intakes of “choline”/“choline and betaine” were not significantly associated with CVD risk; however, the higher intakes of choline and betaine were associated with higher serum concentrations of CRP, IL-6 and TNF-α. Individuals with high plasma choline levels were obese and had elevated plasma triglycerides, HDL and non-HDL cholesterol levels; whereas high plasma betaine levels were inversely associated with these biochemical markers. Both choline and betaine supplementation resulted in increased blood lipid profiles.

CONCLUSION: Although dietary intakes of choline and betaine were not significantly associated with CVD incidence, the long-term consumption of these nutrients have been shown to prevent CVD mortality by decreasing inflammation and other risk factors.

Rajaie, S. and A. Esmaillzadeh, Dietary choline and betaine intakes and risk of cardiovascular diseases: review of epidemiological evidence. ARYA Atheroscler, 2011. 7(2): p. 78-86