Betaine is atheroprotective

Paraoxonase (PON1) is an antioxidant enzyme that prevents LDL oxidation as well as detoxifies homocysteine thiolactone (HCTL), both of which can cause atherosclerosis. Chronic alcohol (ETOH) and high ω-3 polyunsaturated fatty acids (ω-3 PUFA) consumption may affect PON1 status presumably via reactive oxygen species by depleting liver glutathione (GSH), whereas betaine may counter their effects.

Experimental rats belonging to various dietary groups were pair-fed with Lieber-DeCarli low (2.8% the dietary calories as ω3-fatty acids) and high (13.8% the dietary calories as ω3-fatty acids) menhaden fish alcohol-liquid diets with and without betaine (10 g/l diet) for 8 weeks after which liver PON1 mRNA, GSH, lipid score, and serum PON1, HCTLase, and ALT activities were measured.

Betaine restored liver PON1 mRNA expressions in low and high ω-3 PUFA ETOH groups with parallel restorations of PON1 activity and liver GSH. Concomitantly, betaine reduced hepatosteatosis accompanied by alleviation of liver injury caused by chronic alcohol and high ω-3 PUFA.

Dietary betaine was atheroprotective by restoring liver glutathione (GSH) that quenches free radicals, but also may alleviate liver injury by reducing hepatosteatosis.

Varatharajalu et al (2009). "Betaine Protects Chronic Alcohol and Omega-3 PUFA-Mediated Down-Regulations of PON1 Gene, Serum PON1 and Homocysteine Thiolactonase Activities With Restoration of Liver GSH." Alcoholism: Clinical and Experimental Research. Epub Dec 17.

Metabonomics in ulcerative colitis

Nuclear magnetic resonance (NMR) spectroscopy and appropriate multivariate statistical analyses have been employed on mucosal colonic biopsies, colonocytes, lymphocytes, and urine from patients with ulcerative colitis (UC) and controls in order to explore the diagnostic possibilities, define new potential biomarkers, and generate a better understanding of the pathophysiology.

Samples were collected from patients with active UC, quiescent UC, and from controls and analyzed by NMR spectroscopy. Significant differences between controls and active UC were discovered in the metabolic profiles of biopsies and colonocytes. In the biopsies from patients with active UC higher levels of antioxidants and of a range of amino acids, but lower levels of lipid, glycerophosphocholine (GPC), myo-inositol, and betaine were found, whereas the colonocytes only displayed low levels of GPC, myo-inositol and choline. Interestingly, 20% of inactive UC patients had similar profiles to those who were in an active state.

This study demonstrates the possibilities of metabonomics as a diagnostic tool in active and quiescent UC and provides new insight into pathophysiologic mechanisms.

Bjerrum et al (2009). "Metabonomics in ulcerative colitis: diagnostics, biomarker identification, and insight into the pathophysiology." J Proteome Res: Epub Oct.

Betaine treatment of NAFLD

This was a randomized placebo-control study of 55 patients with biopsy-proven NASH who received either oral betaine (20 g daily) or placebo for 12 months. Treatment groups were comparable at baseline. Of the 35 patients (17 betaine, 18 placebo) who completed the study, 34 patients (16 betaine, 18 placebo) underwent posttreatment liver biopsy.

Compared to placebo, betaine improved hepatic steatosis and may protect against worsening steatosis. High-dose betaine supplementation failed to reduce S-adenosylhomocysteine and did not positively affect any of the second hit mechanisms postulated to contribute to NASH.

Abdelmalek et al (2009). "Betaine for nonalcoholic fatty liver disease: Results of a randomized placebo-controlled trial." Hepatology 50(6): 1818-26.

Amniotic fluid metabolites

This study aimed to establish and compare normative metabolite concentrations in 2nd and 3rd trimester human amniotic fluid samples in an effort to reveal metabolic biomarkers of fetal health and development.

21 metabolite concentrations were compared between 2nd and 3rd trimester amniotic fluid samples using (1)H high resolution magic angle spinning (HR-MAS) spectroscopy. 16 of 21 metabolite concentrations differed significantly between 2nd and 3rd trimester groups. Betaine and creatinine concentrations increased significantly, while glucose, citrate, pyruvate, and numerous amino acid concentrations decreased significantly with advancing gestation.

These results provide key normative data for 2nd and 3rd trimester amniotic fluid metabolite concentrations and provide the foundation for future development of magnetic resonance spectroscopy (MRS) biomarkers to evaluate fetal health and development.

Cohn et al (2009). "Quantitative metabolic profiles of 2nd and 3rd trimester human amniotic fluid using (1)H HR-MAS spectroscopy." Magma. Epub Sept.

Obesity, high fat diet, and urine metabolites

To investigate metabolic changes in the urine of a rat model of obesity induced by a high-fat diet (HFD), rats were divided into the following four groups based on the diet type and degree of weight gain: normal-diet (ND) low gainers, ND high gainers, HFD low gainers, and HFD high gainers. Biochemical analyses of visceral fat-pad weight, plasma, and liver tissues were performed.

It was observed that the metabolic profile of urine obtained by 1H-NMR-spectroscopy-based metabolomic analysis differed between ND low gainers and ND high gainers even though these animals consumed the same normal diet. Several key metabolites in urine, such as betaine, taurine, acetone/acetoacetate, phenylacetylglycine, pyruvate, lactate, and citrate contributed to the classification of these two groups. The metabolic profile of urine also differed between ND low gainers and HFD high gainers, which consumed the different diet and showed a different weight gain.

In short, these key metabolites, including betaine, were at higher concentrations in urine for high gainers vs low gainers.

This study has identified features of urine metabolites in various groups and demonstrated the reliability of an NMR-based metabolomics approach to investigate the effects of the diet and the physical constitution on obesity.

Kim et al (2009). "(1)H-nuclear magnetic resonance spectroscopy-based metabolic assessment in a rat model of obesity induced by a high-fat diet." Anal Bioanal Chem. Epub August

Ovarian cancer risk

The authors prospectively examined the relationship between the intake of choline/betaine and ovarian cancer risk among participants from the Nurses' Health Study (NHS) and NHSII. There were no associations between total choline, betaine, as well as choline plus betaine intake and ovarian cancer risk. Results did not vary by alcohol consumption, folate intake or after the exclusion of cases diagnosed during the 4-year period after dietary assessment.

Kotsopoulos et al (2010). "Dietary betaine and choline intake are not associated with risk of epithelial ovarian cancer." Eur J Clin Nutr 64(1): 111-114.

Fibrates may cause abnormal urinary betaine loss

Abnormal urinary loss of betaine is common in patients with the metabolic syndrome or diabetes mellitus. These patients are often treated with fibrates which alter renal function and raise plasma homocysteine concentrations.

- patients taking bezafibrate had higher betaine excretion than patients not taking fibrates
- of 32 patients taking bezafibrate, 20 had abnormal (>97.5 %-ile) betaine excretion
- plasma homocysteine correlated positively with betaine excretion in male patients with lipid disorders who were not taking fibrate, but the relationship was stronger if patients taking bezafibrate were included
- in elderly (>65 years) subjects with hypertension there was a similar correlation, which was stronger when a subject taking bezafibrate was included

Abnormal betaine excretion is common in patients treated with bezafibrate. Bezafibrate appears to exacerbate betaine loss, which will cause a rise in plasma homocysteine. Betaine supplementation could be considered in conjunction with fibrate therapy.

Lever et al (2009). "Fibrates may Cause an Abnormal Urinary Betaine Loss Which is Associated with Elevations in Plasma Homocysteine." Cardiovasc Drugs Ther. Epub Aug 4

Prevention of amyloid β-peptide formation

S-Adenosylhomocysteine (SAH) has been implicated as a risk factor for neurodegenerative diseases such as Alzheimer's disease. SAH is a potent inhibitor of all cellular methyltransferases and this study showed that SAH increased amyloid β-peptide (Aβ) formation in a concentration-dependent manner (10–500 nM), and this effect of SAH was accompanied by significantly increased expression of APP and PS1 proteins.

Pre-incubation of cells with betaine (1.0 mM), 30 min followed by incubation with SAH (500 nM) or 5′-azc (20 μM) for 24 h markedly prevented the expression of Aβ protein (by 50%, P < 0.05) and the gene promoter hypomethylation of APP and PS1.

Lin et al (2009). "S-Adenosylhomocysteine increases beta-amyloid formation in BV-2 microglial cells by increased expressions of beta-amyloid precursor protein and presenilin 1 and by hypomethylation of these gene promoters." Neurotoxicology 30(4): 622-7.

Protective effect in liver cancer

Four groups of rats were given diethylinitrosamine (DEN) and fed with AIN-93G diets supplemented with 0, 10, 20 or 40 g betaine/kg (model, 1%, 2%, and 4% betaine, respectively), while the control group, received no DEN, fed with AIN-93G diet. Eight or 15 weeks later, the expression of p16 and c-myc mRNA was examined by Real-time PCR (Q-PCR). The DNA methylation status within the p16 and c-myc promoter was analyzed using methylation-specific PCR.

Compared with the model group, numbers and areas of glutathione S-transferase placental form (GST-p)-positive foci were decreased in the livers of the rats treated with betaine (P < 0.05). Although the frequency of p16 promoter methylation in livers of the four DEN-fed groups appeared to increase, there is no difference among these groups after 8 or 15 weeks (P > 0.05). Betaine supplementation attenuated the down-regulation of p16 and inhibited the up-regulation of c-myc induced by DEN in a dose-dependent manner (P < 0.01). Meanwhile, increases in levels of malondialdehyde (MDA) and glutathione S-transferase (GST) in model, 2% and 4% betaine groups were observed (P < 0.05). Finally, enhanced antioxidative capacity (T-AOC) was observed in both the 2% and 4% betaine groups.

The data suggest that betaine attenuates DEN-induced damage in rat liver and reverses DEN-induced changes in mRNA levels.

Du et al (2009). "Assessment of the effect of betaine on p16 and c-myc DNA methylation and mRNA expression in a chemical induced rat liver cancer model." BMC Cancer 9(1): 261.

Reduced breast cancer risk and mortality

The authors investigated the associations of dietary intakes of choline and betaine and breast cancer risk and mortality in the population-based Long Island Breast Cancer Study Project.

- there was an indication that a higher intake of free choline was associated with reduced risk of breast cancer
- higher intakes of betaine, phosphocholine, and free choline were associated with reduced all-cause as well as breast cancer-specific mortality in a dose-dependent fashion
- the betaine-homocysteine methyltransferase gene (BHMT) rs3733890 polymorphism was associated with reduced breast cancer-specific mortality

The study suggests that high intake of betaine and choline may be a promising strategy to prevent the development of breast cancer and to reduce its mortality.

Xu et al (2009). "High intakes of choline and betaine reduce breast cancer mortality in a population-based study". FASEB J., epub July 27.

Betaine content of soy sauce

Soy sauce consists of many metabolites that are produced during fermentation or aging and that have various health benefits. However, their comprehensive assessment has been limited. This paper presents a metabolic characterization of soy sauce, especially that aged up to 12 years using 1H NMR spectroscopy coupled with multivariate pattern recognition techniques.

Elevated amino acids and organic acids and the consumption of carbohydrate were associated with continuous involvement of microflora in aging for 12 years. In particular, continuous increases in the levels of betaine were found during aging for up to 12 years, demonstrating that microbial- or enzyme-related metabolites were also coupled with osmotolerant or halophilic bacteria present during aging.

Ko et al (2009). "Metabolomic Insight into Soy Sauce through (1)H NMR Spectroscopy." J Agric Food Chem. Epub Jul 10

One carbon metabolism and NTD

Folic acid is known to reduce risk of neural tube defects (NTDs). Even so, NTDs continue to occur despite individual supplementation or population fortification with folic acid. This study investigated other nutrients related to one-carbon metabolism that may affect NTD risk.

This prospective study included data from more than 180,000 pregnant women in California from 2003 through 2005. Midpregnancy serum specimens were linked with delivery information regarding the presence of a NTD, another structural malformation, or no malformation in the fetus. They identified 80 NTD-affected pregnancies (cases) and we randomly selected 409 pregnancy controls. Serum specimens were tested for methylmalonic acid, homocysteine, cysteine, methionine, total choline, betaine, cystathionine, vitamin B6, folate, vitamin B12, riboflavin, and creatinine.

Elevated NTD risks associated with lower levels of total choline, and reduced risks with higher levels of choline. There were no differences between cases and controls for any other analytes.

This is the first study to investigate total choline in NTD-affected pregnancies. The findings for choline, for which low levels were a risk factor and higher levels were a protective factor for NTDs, may offer a useful clue toward understanding the complex etiologies of NTDs in an era of folic acid fortification of the food supply.

Shaw et al (2009). "Choline and Risk of Neural Tube Defects in a Folate-Fortified Population." Epidemiology. epub July 10 2009

Plasma betaine increases upon intake of high-fiber rye buns

An NMR-based metabonomic study explored the biochemical effects of a rye based fiber-rich diet in hypercholesterolemic pigs.

The pigs were fed high-fat, high-cholesterol rye- (n = 9) or wheat- (n = 8) based buns with similar levels of dietary fiber for 9-10 wk. Fasting plasma samples were collected 2 days before and after 8 and 12 days on the experimental diets, while postprandial samples taken after 58-67 days, and 1H NMR spectra were acquired on these.

The NMR spectra demonstrated a high intensity for the spectral region at 3.29 ppm in the rye diet. The 3.29 ppm signal is ascribed to N(CH3)3 protons in betaine, which may be an important contributor to the health promoting effects of rye.

Bertram et al (2009). "NMR-based metabonomics reveals that plasma betaine increases upon intake of high-fiber rye buns in hypercholesterolemic pigs." Mol Nutr Food Res. epub 14 July 2009

Creatine synthesis and BHMT

This study investigated the:

- rate of creatine accretion by the neonatal piglet
- sources of this creatine
- activities of the enzymes of creatine synthesis
- burden that endogenous creatine synthesis places on the metabolism of the 3 amino acids required for this synthesis: glycine, arginine, and methionine.

They found that:

- piglets acquire 12.5 mmol of total creatine (creatine plus creatine phosphate) between 4 and 11 d of age
- 1/4 of creatine accretion in neonatal piglets may be provided by sow milk and 3/4 by de novo synthesis by piglets
- several enzyme activities related to creatine synthesis increased, including betaine:homocysteine methyltransferase in the liver.

They concluded that creatine synthesis is a quantitatively major metabolic process in piglets.

Brosnan et al (2009). "Creatine synthesis is a major metabolic process in neonatal piglets and has important implications for amino acid metabolism and methyl balance." J Nutr 139(7): 1292-7.

Developing fatty liver

The authors evaluated the effects of betaine supplementation on goose liver weight, liver/body weight, serum parameters and morphological changes.

Compared with the control and overfed groups, the geese that were fed the betaine diet showed increased liver weight and decreased abdominal adipose tissue weight compared with the overfeeding groups. Betaine treatment also significantly increased ChE, HDL, LDH and ALT levels. Decreased macrovesicular steatosis and increased microvesicular steatosis were observed in the betaine-treated group, and the lipid was well-distributed in the betaine supplement group. The expression of S14α mRNA in the livers of the betaine-treated geese was higher than that in the control or the overfed geese. The DNA methylation pattern in the S14α gene transcription start site may not be related to the expression of S14α transcript in response to betaine supplementation.

Su et al (2009). "The effects of dietary betaine supplementation on fatty liver performance, serum parameters, histological changes, methylation status and the mRNA expression level of Spot14[alpha] in Landes goose fatty liver." Comparative Biochemistry and Physiology - Part A: Molecular & Integrative Physiology. Epub May

Improved growth performance

120 female pigs were fed either a control commercial diet or the control diet supplemented with 2, 4 and 6% betaine for 31 days. Pigs fed betaine had:

- lower average daily feed intake (ADFI) (for 2% diet)
- higher average daily gain (ADG)
- lower feed conversion ratio (FCR)
- increased loin CIE a* (redness)
- higher loin shear force value
- decreased total blood cholesterol concentrations
- increased saturated fatty acid and decreased unsaturated fatty acid levels in muscle
- increased betaine concentrations in the loin muscle

It was concluded that dietary betaine supplementation of finishing pigs can improve growth performance, reduce blood cholesterol concentrations, and produce detectable betaine concentrations in the lion muscle.

Yang et al (2009). "Effects of dietary glycine betaine on growth and pork quality of finishing pigs." Asian-Australasion J Animal Sci 22(5): 706.

5,10-Methylenetetrahydrofolate reductase deficiency

Betaine therapy was given for 2 years to a 2-year-old boy with 5,10-methylenetetrahydrofolate reductase deficiency. Used as a methyl donor to lower homocysteine levels through methylation of methionine, betaine has been reported to be effective in treating homocystinuria. Satisfactory biochemical and clinical responses were obtained with the following regimen: betaine started in the newborn period at increasing doses to reach 1 g given six times a day. It is suggested that frequent administration of a moderate dose may provide clinical and biochemical benefit.

Ucar et al (2009). "Titration of betaine therapy to optimize therapy in an infant with 5,10-methylenetetrahydrofolate reductase deficiency." Eur J Pediatr. Epub May 17

One Carbon Metabolites and Prostate Cancer Risk

This study investigated components of one−carbon metabolism in relation to prostate cancer risk in a large prospective epidemiological study. Circulating plasma concentrations of betaine, cysteine, methionine, and vitamin B6 were not associated with prostate cancer risk, whereas elevated concentrations of choline and vitamin B2 may be associated with an increased risk.

Johansson et al (2009). "One-Carbon Metabolism and Prostate Cancer Risk: Prospective Investigation of Seven Circulating B Vitamins and Metabolites." Cancer Epidemiol Biomarkers Prev 18(5): 1538-1543.

Characterization of industrial rye and wheat brans

Six different rye brans from Sweden, Denmark and Finland were analysed and compared with two wheat brans regarding colour, particle size distribution, microscopic structures and chemical composition including proximal components, vitamins, minerals and bioactive compounds.

There were many variations within rye brans, and differences from wheat bran. Rye bran had less betaine (194-278 mg/100 g) than wheat brans (431-441 mg/100 g).

Kamal-Eldin et al (2009). "Physical, microscopic and chemical characterisation of industrial rye and wheat brans from the Nordic countries." Food Nutr Res 53 DOI: 10.3402/fnr.v53i0.1912.

Alcoholic Liver Disease and Methionine Metabolism

This review states that, "Of all the therapeutic modalities that are presently being used to attenuate ethanol-induced liver injury, betaine has been shown to be the most effective in a variety of experimental models of liver disease. Betaine, by virtue of aiding in the remethylation of homocysteine, removes both toxic metabolites (homocysteine and S-adenosylhomocysteine), restores S-adenosylmethionine level, reverses steatosis, prevents apoptosis and reduces both damaged protein accumulation and oxidative stress. Thus, betaine is a promising therapeutic agent in relieving the methylation and other defects associated with alcoholic abuse."

Kharbanda, K. K. (2009). Alcoholic liver disease and methionine metabolism. Semin Liver Dis, 29(2), 155-165.

Betaine prevents Mallory-Denk body formation

Drug-primed mice were fed betaine, together with DDC, which was refed for 7 days. Betaine:

- reduced Mallory-Denk body (MDB) formation (markers for liver disease)
- decreased the liver/body weight ratio
- decreased the number of FAT10 positive liver cells
- prevented the decreased expression of BHMT, AHCY, MAT1a and GNMT and the increased expression of MTHFR, caused by DDC refeeding
- prevented the rise in S-Adenosylhomocysteine (SAH) levels caused by DDC refeeding

Oliva et al (2009). "Betaine prevents Mallory-Denk body formation in drug-primed mice by epigenetic mechanisms." Exp Mol Pathol 86(2): 77-86.

Reducing IR and NAFLD

This is an abstract from the 2008 AASLD meeting. Rats were fed one of 3 diets for 8 months:

- standard chow (SF) (9% cal for fat)
- high fat (HF) (20% cal from fat)
- HF plus betaine (HF+B) (1.5% betaine in drinking water)

In this high fat diet-induced model of insulin resistance and NAFLD, oral betaine:

- reduced fasting insulin and glucose
- reversed hepatic steatosis
- increased activation of the insulin signaling pathway
- reversed hepatic insulin resistance, as demonstrated by reduced gluconeogenesis and increased glycogen content

Kathirvel et al (2008). "Betaine reduces insulin resistance and hepatic steatosis, and augments hepatic signaling, in an animal model of NAFLD." Hepatology 48(S1): 408A-505A.

Betaine and homocysteine

In a randomized crossover study, 8 healthy males (22–36 y) consumed either a betaine-rich diet (800 mg/day) or a betaine supplement (0.5 g twice daily) for 14 days. Fasting blood samples were collected on day −5, −1 (pre-treatment) 0, 2, 6, 9, 13 (treatment), 14 and 18 (post-treatment). Post-methionine load blood samples were collected on day −5, 0, 6 and 13, while 24 h urine samples were collected on day −5, 0, 6, 13 and 14. Plasma betaine, dimethylglycine, homocysteine and urine betaine, dimethylglycine and creatinine concentrations were measured.

Plasma betaine concentrations significantly increased for both treatments compared to pre-treatment values (P < 0.001). Fasting homocysteine levels were minimally affected. Both treatments reduced post-methionine load homocysteine and this effect tended to be greater following a betaine-rich diet (P = 0.108). Small increases in urinary betaine excretion were observed following both treatments (≈1.5% of supplement; ≈1.3% of dietary betaine). Most was attributable to increased excretion of betaine as dimethylglycine.

Therefore, supplemental or dietary betaine similarly increase circulating betaine concentrations and attenuate the post-methionine load rise in homocysteine concentrations.

Atkinson et al (2009). "Dietary and supplementary betaine: Effects on betaine and homocysteine concentrations in males." Nutr Metab Cardiovasc Dis. Apr 3 Epub.

One carbon metabolism and birth defects

The objective was to investigate whether intake of nutrients involved in one-carbon metabolism (folate, vitamin B6, vitamin B12, riboflavin, choline, betaine, zinc, and methionine) through diet alone or in combination with a supplement containing folic acid influenced the risk for transverse limb deficiency (TLD) and longitudinal limb deficiency (LLD).

They analyzed 1997-2003 data from the National Birth Defects Prevention Study and included 324 case infants with TLD, 158 case infants with LLD, and 4982 nonmalformed control infants.

TLD and LLD were not associated with supplement use, but TLD was associated with low intakes of riboflavin from diet.

Robitaille et al (2009). "Maternal nutrient intake and risks for transverse and longitudinal limb deficiencies: Data from the National Birth Defects Prevention Study, 1997-2003." Birth Defects Research Part A: Clinical and Molecular Teratology. March 1 Epub.

Effect of casein diet on hepatic BHMT and betaine

Study 1 - male Wistar rats were fed diets differing in casein level from 5 to 50% for 14 d. Plasma total homocysteine concentration was positively correlated with dietary casein level in the range of 5 to 10% but inversely correlated with dietary casein level in the range of 10 to 50%. Hepatic cystathionine β-synthase (CBS) and betaine-homocysteine S-methyltransferase (BHMT) activities and renal CBS activity increased in response to dietary casein level in the range of 10 to 50%, whereas hepatic serine and betaine concentrations decreased with increasing dietary casein levels.

Study 2 - rats were fed a 10% casein diet or 10% casein+17.2% amino acid mixture diet for 14 d. Plasma homocysteine concentration was significantly lower in rats fed the amino acid mixture-added diet than in rats fed the 10% casein diet, indicating that the hypohomocysteinemic effect of high casein diets was elicited by amino acids, not by casein contaminants.

Study 3 - the degree of increase in plasma homocysteine concentration caused by dietary supplementation with 0.75% L-methionine was significantly lower in rats fed the 40% casein diet than in rats fed the 10% casein diet.

These results indicate that high casein diets do not increase but rather decrease plasma homocysteine concentration and cause resistance to hyperhomocysteinemic treatment, and suggest that such effects of high casein diets are mediated at least by increased activities of CBS and BHMT.

Ohuchi et al (2009). "High casein diet decreases plasma homocysteine concentration in rats." J Nutr Sci Vitaminol (Tokyo) 55(1): 22-30.

Protection against myocardial infarction

This study investigated the protective effect of betaine on changes in the levels of membrane-bound ATPase activities, lipid peroxidation, sulfhydryl activities, and mineral status in isoprenaline-induced myocardial infarction in Wistar rats, an animal model of myocardial infarction in man. Oral administration of betaine (250 mg/kg body weight/day for a period of 30 days) significantly (p < 0.05) improved the following isoprenaline-induced abnormalities:

- maintained levels of sodium, potassium, and calcium in plasma and heart tissue
- protected membrane-bound ATPase in the heart tissue
- preserved myocardial sulfhydryl activities
- counteracted lipid peroxidation

Ganesan et al (2009). "Protective effect of betaine on changes in the levels of membrane-bound ATPase activity and mineral status in experimentally induced myocardial infarction in Wistar rats." Biological Trace Element Research. April 8 Epub

Osmolyte protection against urea denaturation of chymotrypsin

The destabilizing effect of urea on proteins can be counteracted by methylamines (osmolytes), such as trimethylamine N-oxide (TMAO), betaine, and sarcosine. This study compared the counteracting effects of these methylamines on urea-induced denaturation of α-chymotrypsin (CT). They measured the hydrodynamic diameter (dH) and the thermodynamic properties (Tm, ΔH, ΔGU, and ΔCp) with dynamic light scattering (DLS) and differential scanning calorimeter (DSC), respectively. The osmolytes strongly counteracted the urea actions on α-chymotrypsin, with TMAO having the strongest effect.

Venkatesu et al (2009). "Osmolyte Counteracts Urea-Induced Denaturation of alpha-Chymotrypsin." The Journal of Physical Chemistry B 113(15): 5327-5338.

Moderate changes in dietary choline/betaine intake and blood indicators of status

Healthy premenopausal women (n=45, 18–46 years) with the MTHFR 677CC (n=28) or TT (n=17) genotype consumed a folate-restricted diet for 2 weeks followed by randomization to one of four dietary treatments (n=6–9/group) differing in total choline (344–486 mg/day), betaine (122–349 mg/day) and/or folate (400–800 μg dietary folate equivalents/day) content for 12 weeks.

No significant changes were detected in the measured variables (plasma levels of choline moieties (i.e., betaine, choline, phosphatidylcholine and sphingomyelin) and/or leukocyte global DNA methylation) in response to dietary increases in choline (i.e., 41% increase) or betaine (i.e., 286% increase) intake between pretreatment (Week 2) and posttreatment (Week 14) values. However, the MTHFR C677T genotype, alone or together with a diet, influenced betaine (P=.03) and phosphatidylcholine (P=.03).

These data suggest that choline/betaine status is not a reliable indicator of moderate changes in dietary choline/betaine intake possibly due to the engagement of compensatory mechanisms. In addition, the MTHFR C677T genotype appears to influence the direction and use of choline moieties in this group of women.

Abratte et al (2009). "Choline status is not a reliable indicator of moderate changes in dietary choline consumption in premenopausal women." J Nutr Biochem 20(1): 62-9.

Protection against myocardial infarction

This study investigated the protective effect of betaine on changes in the levels of lysosomal enzymes and lipid peroxidation in isoprenaline-induced myocardial infarction in Wistar rats, an animal model of myocardial infarction in man. Male albino Wistar rats were pretreated with betaine (250 mg/kg body weight) daily for a period of 30 days. After the treatment period, isoprenaline (11 mg/100 g body weight) was intraperitoneally administered to rats at intervals of 24 h for 2 days.

In isoprenaline-injected rats, the activities of plasma lysosomal enzymes (beta-glucuronidase, beta-galactosidase, beta-glucosidase, and acid phosphatase) increased significantly (p < 0.05), but activities decreased in heart tissue. Also, the level of lipid peroxidation was higher in heart lysosomes of isoprenaline-injected rats. Pretreatment with betaine prevented the changes in the activities of these lysosomal enzymes. Thus, the results show that betaine protects the lysosomal membrane against isoprenaline-induced myocardial infarction. The observed effects might be due to the free radical-scavenging and membrane-stabilizing properties of betaine.

Ganesan and Anandan (2009). "Protective effect of betaine on changes in the levels of lysosomal enzyme activities in heart tissue in isoprenaline-induced myocardial infarction in Wistar rats." Cell Stress Chaperones. March 18 Epub.

Greater daily gain and lean deposition in pigs

This study investigated the interactions between dietary ractopamine and betaine on growth and carcass characteristics in restrictively fed pigs. Pigs fed betaine had:

- greater daily gain (+8%)
- greater lean deposition (+5%)
- no effect on fat deposition

Dunshea et al (2009). "Dietary betaine and ractopamine combine to increase lean tissue deposition in finisher pigs, particularly gilts." Animal Production Science 49(1): 65-70.

Effect of pH on protein stabilizing property of betaine

This study investigated the pH dependence of the stabilizing effect of betaine on three different proteins - α-lactalbumin (α-LA), lysozyme and ribonuclease-A (RNase-A). They measured Tm (midpoint of denaturation), ΔHm (denaturational enthalpy change at Tm), ΔCp (constant-pressure heat capacity change) and ΔGDo (denaturational Gibbs energy change at 25 °C) of proteins in the presence of different betaine concentrations.

They found that betaine:

- stabilizes RNase-A at all pH values
- has opposite effects on α-LA and lysozyme at high pH and low pH values
- did not significantly change ΔHm and ΔCp

Singh et al (2009). "Glycine betaine may have opposite effects on protein stability at high and low pH values." Biochim Biophys Acta. Epub

Betaine reduces athersclerosis and inflammatory response

This study investigated the effect of betaine supplementation on atherosclerotic lesion in apolipoprotein (apo) E-deficient mice. Four groups of these mice were fed AIN-93G diets supplemented with 0, 1, 2, or 4 g betaine/100 g diet (no, 1, 2, and 4% betaine, respectively). Wild-type C57BL/6 J mice were fed AIN-93G diet (wild-type). Mice were sacrificed after 0, 7, or 14 weeks of the experimental diets. Atherosclerotic lesion area in the aortic sinus, levels of tumor necrosis factor (TNF)-α and monocyte chemoattractant protein (MCP)-1 in aorta and serum, serum lipids, and methylation status of TNF-α promoter in aorta were determined.

The results showed:

- compared with no-betaine mice after 14 weeks, mice receiving 1%, 2%, or 4% betaine had 10.8, 41, and 37% smaller lesion area, respectively.
- betaine supplementation reduced aortic expression of TNF-α in a dose-dependent way.
- betaine supplementation for 14 weeks led to higher concentrations of serum total cholesterol (P < 0.01), LDL cholesterol (P < 0.05), and lower body weight (P < 0.05).

The authors concluded that despite exacerbating hyperlipidemia in apoE-deficient mice, betaine may exert its anti-atherogenic effect by inhibiting aortic inflammatory response mediated by TNF-α.

Lv et al (2009). "Betaine supplementation attenuates atherosclerotic lesion in apolipoprotein E-deficient mice." Eur J Nutr 48(4): 205-12.

Muscle endurance and quality of repetitions

This study examined the efficacy of 15 days of betaine supplementation on muscle endurance, power performance and rate of fatigue in active college-aged men. Male subjects (24) were randomly assigned to one of two groups; the first consumed betaine daily (2.5 g/d), and the second consumed a placebo. Subjects were tested prior to the onset of supplementation (T1) and 7 (T2) and 14 days (T3) following supplementation. Subjects were tested over a 2-day period for:

- vertical jump power (VJP)
- bench press throw (BPT) power
- number of squat repetitions at 75% 1-RM (peak and mean power)
- number of bench press repetitions at 75% 1-RM (peak and mean power)
- two 30-sec Wingate anaerobic power tests (WAnT) (Day 2)

There were no differences in power assessment (VJP, BPT, WAnT) between the groups, or in the repetitions performed to exhaustion or in the number of repetitions performed at 90% of both peak and mean power between the groups in the bench press exercise.

However, the number of repetitions performed in the squat exercise for BET was significantly greater (p < 0.05) than that seen for PL at T2. The number of repetitions performed at 90% or greater of peak power in the squat exercise was significantly greater for BET at both T2 and T3 than PL. It was concluded that either 1 or 2 weeks of betaine supplementation in active, college males improved muscle endurance of the squat exercise, and increased the quality of repetitions performed.

Hoffman et al (2009). "Effect of Betaine Supplementation on Power Performance and Fatigue." Journal of the International Society of Sports Nutrition 6(1): 7.

Liver proteomics

A proteomic profiling strategy examined the effects of ethanol and betaine diet supplementation on major liver protein level changes. Male Wistar rats were fed control, ethanol or betaine supplemented diets for 4 weeks. Livers were removed and liver cytosolic proteins resolved by one-dimensional and two-dimensional separation techniques.

In rats fed the betaine supplemented ethanol diet, there was significant upregulation of:

- betaine homocysteine methyltransferase-1
- methionine adenosyl transferase-1
- glycine N-methyltransferase

The authors hypothesise that this concerted upregulation of these methionine metabolic pathway enzymes is the protective mechanism by which betaine restores a normal metabolic ratio of liver S-adenosylmethionine to S-adenosylhomocysteine. Ethanol also induced significant downregulation of carbonic anhydrase-III protein levels which was not restored by betaine supplementation. Carbonic anhydrase-III can function to resist oxidative stress, and the authors hypothesise that carbonic anhydrase-III protein levels compromised by ethanol consumption, contribute to ethanol-induced redox stress.

Kharbanda et al (2009). "Proteomics reveal a concerted upregulation of methionine metabolic pathway enzymes, and downregulation of carbonic anhydrase-III, in betaine supplemented ethanol-fed rats." Biochem Biophys Res Commun 381(4): 523-7.

Betaine intake measurement

The three-year reliability of reported dietary intake was similar for betaine and related nutrients, in the range as that published in the literature for other micronutrients. The mean intake of betaine was estimated at 106 mg/d.

Bidulescu et al (2009). "Repeatability and measurement error in the assessment of choline and betaine dietary intake: the Atherosclerosis Risk in Communities (ARIC) Study." Nutrition Journal 8(1): 14.

Betaine antioxidant effect protects against liver damage

This study showed that betaine may protect liver from fibrogenesis by maintaining cellular antioxidant capacity.

Male rats were supplemented with betaine (1%, w/v) in drinking water from 2 weeks prior to the initiation of dimethylnitrosamine (DMN) treatment (10mg/(kg day), i.p., 3 days/week, for 1, 2, or 4 weeks). Induction of liver injury was determined by quantifying serum alanine aminotransferase, aspartate aminotransferase activities, bilirubin levels, hepatic xenobiotic-metabolizing capacity, histopathological changes and 4-hydroxyproline levels. Development of oxidative injury was estimated by malondialdehyde (MDA) levels and total oxyradical scavenging capacity (TOSC) of liver and serum toward hydroxyl, peroxyl radicals, and peroxynitrite.

Betaine supplementation markedly attenuated the DMN-induced hepatotoxicity, fibrosis and oxidative injury (e.g. elevation of MDA and the reduction of TOSC were depressed significantly).

Kim et al (2009). "Alleviation of dimethylnitrosamine-induced liver injury and fibrosis by betaine supplementation in rats." Chem Biol Interact 177(3): 204-11.

Individuality of plasma and urine betaine and DMG

The individuality (within subject consistency) of plasma and urine betaine and N,N-dimethylglycine (DMG) was compared in two groups of 8 males (ages 19 to 40) either over a single day or over an 8 week period. The study found that plasma betaine and urinary betaine excretions are more individual than DMG, and that plasma and urine betaine are highly individual in the general population.

Lever et al (2009). "Plasma and urine betaine and dimethylglycine variation in healthy young male subjects." Clin Biochem 42: 706-12.

Papers 2004-7

Lever et al (2004). "Short and long-term variation of plasma glycine betaine concentrations in humans." Clin Biochem 37(3): 184-90.

Slow et al (2004). "Betaine analogues alter homocysteine metabolism in rats." Int J Biochem Cell Biol 36(5): 870-80.

Lever et al (2005). "Homocysteine, glycine betaine, and N,N-dimethylglycine in patients attending a lipid clinic." Metabolism - Clinical and Experimental 54(1): 1-14.

Slow et al (2005). "The betaine content of New Zealand foods and estimated intake in the New Zealand diet." J Food Comp Anal 18: 473-85.

Lever et al (2007). "An abnormal urinary excretion of glycine betaine may persist for years." Clin Biochem 40(11): 798-801.

Lever et al (2007). "Inter- and intra-individual variations in normal urinary glycine betaine excretion." Clin Biochem 40(7): 447-53.

Lever et al (2007). "Sex differences in the control of plasma concentrations and urinary excretion of glycine betaine in patients attending a lipid disorders clinic." Clin Biochem 40(16-17): 1225-31.

Atkinson et al (2008). "Dietary and supplementary betaine: acute effects on plasma betaine and homocysteine concentrations under standard and postmethionine load conditions in healthy male subjects." Am J Clin Nutr 87(3): 577-585.

Betaine and choline content of wheat fractions

This present study suggests that the wheat aleurone layer contains the greatest concentration of both betaine and choline (1553 and 210 mg/100 g of sample, respectively). The bran fraction contained 867 and 102 mg/100 g of sample of betaine and choline, respectively, while the flour fraction contained 23 mg/100 g of sample (betaine) and 28 mg/100 g of sample (choline). The betaine content for the bran was lower, and the choline content was higher compared to previous studies, although it is known that there is large variation in betaine and choline contents between wheat cultivars. The study further emphasizes the superior phytonutrient composition of the aleurone layer.

Graham et al (2009). "Analysis of Betaine and Choline Contents of Aleurone, Bran, and Flour Fractions of Wheat (Triticum aestivum L.) Using 1H Nuclear Magnetic Resonance (NMR) Spectroscopy." J Ag Food Chem 57(5): 1948-1951.

VLDL secretion and hepatic fat export

Chronic ethanol consumption impairs phosphatidyl choline (PC) generation via the phosphatidylethanolamine methyltransferase (PEMT) pathway. This results in diminished very low-density lipoprotein (VLDL) secretion which contributes to the development of hepatic steatosis.

The VLDL secretion in rats fed with control, ethanol and the betaine supplemented diets was determined using Triton WR-1339 to inhibit plasma VLDL metabolism.

Betaine feeding resulted in increased VLDL production rates and fat export from the liver by increasing PEMT-mediated PC generation. This attenuated the development of alcoholic fatty liver.

Kharbanda et al (2009). "Betaine administration corrects ethanol-induced defective VLDL secretion." Mol Cell Biochem. Feb 19 epub.

Rett Syndrome

A 12-month, double-blind, placebo-controlled folate–betaine trial studied 68 females with Rett syndrome. Participants were randomized as young (< age 5 years) or old ( age 5 years). Structured clinical assessments occurred at baseline, 3, 6, and 12 months. Primary outcome measures included quantitative evaluation of breathing and hand movements during wakefulness, growth, anthropometry, motor/behavioral function, and qualitative evaluations from electroencephalograms and parent questionnaires.

Objective evidence of improvement was not found, but subjective improvement from parent questionnaires was noted for the <5 years group.

Glaze et al (2009). "A Study of the Treatment of Rett Syndrome With Folate and Betaine." J Child Neurol. Feb 18 epub.

Betaine inhibits atherosclerosis via anti-inflammation

Five groups of mice were studied: ApoE-deficient (model group and three betaine groups) and wild-type mice as control. The control group and model group were fed AIN-93G diet. Three betaine groups were fed AIN-93G diet supplemented with 1, 2, 4 g betaine/100 g diet, respectively.

The study found:

- The percentage of aorta sinus plaque to lumen area of 1% and 2% betaine groups were 41% and 33% smaller than that of the model group.

- Serum TNF-alpha level of three betaine groups were lower than that of the model group, but there was no significant difference in the methylation status of TNF-alpha promotor among all five groups.

They concluded that betaine could inhibit the development of atherosclerosis via anti-inflammation.

Fan et al (2008). "Anti-atherosclerotic effect of betaine in apolipoprotein E-deficient mice." Zhonghua Yu Fang Yi Xue Za Zhi 42(10): 742-7.

Betaine overcomes perturbations caused by choline deficiency

Rats were fed either a standard (25%) casein or soybean protein diet, however both diets were choline-deprived. The study found that:

- Hyperhomocysteinemia induced by choline deprivation was effectively suppressed by choline, betaine or methionine supplementation.

- Hepatic SAM:SAH ratio was improved.

They concluded that this might be a useful model for investigating the role of betaine in the regulation of plasma homocysteine concentration.

Setoue et al (2008). "Choline deprivation induces hyperhomocysteinemia in rats fed low methionine diets." J Nutr Sci Vitaminol (Tokyo) 54(6): 483-90.

Betaine reduces oxidative stress and inflammation

This study investigated the effects of betaine on the inflammatory process and the oxidative stress in rats with nonalcoholic steatohepatitis (NASH) using control, model, low and high dose betaine groups. Except control group, all rats were fed with high fat diet containing propylthioracil (PTU), a fatty liver-inducing drug, to construct NASH model, and those in low (200 mg/kg) and high (400 mg/kg) dose group received betaine solution intragastric gavage.

Compared with control group, model group rats had:

- increased liver damage
- enhanced expression of tumor necrosis factor-α (TNF-α), inflammatory factor interferon-γ (IEN-γ), and cytochrome-related factor CYP2E1 mRNA in liver
- decreased expression of IL-10 mRNA
- increased levels of malonaldehyde (MDA) and nitrogen monoxide (NO) in liver
- no significant change in cytochrome-related factor CYP3A2

Compared with that in the model group, high and low dose betaine groups had:

- less liver damage
- attenuated expression of TNF-α, TEN-γ, CYP2E1 and CYP3A2 mRNA in liver
- enhanced expression of IL-10 mRNA
- decreased levels of MDA and NO in liver

There was a significant difference in the above parameters between the two betaine treatment groups. Betaine has protective effects on liver injury in rats induced by a high fat diet. The mechanisms may involve improvement in oxidative stress and suppressed expression of inflammatory factors.

Zhang et al (2008). "Effects of betaine on oxidative stress and inflammatory factors in rats with nonalcoholic steatohepatitis." Wuhan Daxue Xuebao (Yixue Ban) 29(5): 587-91.

Betaine alleviates perturbations of a high fat diet

Male rats were provided with a standard liquid diet, a high-fat liquid diet (HF), or a HF diet supplemented with betaine (1%) for 3 wk.

HF diet intake:

- elevated hepatic triglyceride and serum tumor necrosis factor (TNF)
- reduced antioxidant capacity of liver cytosol against hydroxyl and peroxyl radicals
- decreased hepatic S-adenosylmethionine (SAM) and glutathione (GSH)
- increased hypotaurine and taurine concentrations
- depressed methionine adenosyltransferase (MAT) activity
- elevated activity and concentration of cysteine dioxygenase and GSH S-transferase

Betaine supplementation of the HF diet:

- inhibited hepatic fat accumulation and serum TNF elevation
- prevented the decrease in cytosolic antioxidant capacity
- induced MAT activity and concentration
- increased hepatic SAM and GSH
- depressed elevation of hypotaurine and taurine

The results indicate that the metabolism of S-containing substances is significantly disturbed by the HF diet, suggesting a causal role of impairment of hepatic transsulfuration reactions in NAFLD. Betaine supplementation protects the liver from nonalcoholic steatosis and oxidative stress most probably via its effects on the transsulfuration reactions.

Kwon et al (2009). "Impaired Sulfur-Amino Acid Metabolism and Oxidative Stress in Nonalcoholic Fatty Liver Are Alleviated by Betaine Supplementation in Rats." J. Nutr. 139(1): 63-68.

Review: Current status of therapy in NAFLD

This article discusses some of the basic understanding of non alcoholic fatty liver disease (NAFLD), reviews the currently tested therapies (including betaine), some novel therapies, and potential future therapeutic options.

McNear and Harrison (2009). "Review: Current status of therapy in nonalcoholic fatty liver disease." Therapeutic Advances in Gastroenterology 2(1): 29-43.