Homocysteine

1. J Nutr. 2003 May;133(5):1291-5.

Betaine supplementation lowers plasma homocysteine in healthy men and women.

Steenge GR, Verhoef P, Katan MB.

Wageningen Centre for Food Sciences, Nutrition and Health Programme, Wageningen, The Netherlands.

Elevated levels of plasma total homocysteine are associated with a higher risk of cardiovascular disease. Betaine and 5-methyltetrahydrofolate can remethylate homocysteine into methionine via independent reactions. We determined the effect of daily betaine supplementation, compared with both folic acid and placebo, on plasma concentrations of total homocysteine after an overnight fast and after methionine loading in men and women with mildly elevated homocysteine. Groups of twelve subjects ingested 6 g betaine, 800 micro g folic acid with 6 g placebo or 6 g placebo each day for 6 wk. A methionine-loading test (i.e., ingestion of 100 mg L-methionine/kg body mass) was performed before and after 6 wk of supplementation. Fasting plasma homocysteine decreased by 1.8 micro mol/L (95% confidence interval [CI]: -3.6, 0.0, P < 0.05) in the betaine group and by 2.7 micro mol/L (95% CI: -4.5, -0.9, P < 0.05) in the folic acid group. These changes are relative to the change in the placebo group, in which fasting plasma homocysteine rose by 0.5 micro mol/L. Furthermore, betaine suppressed the total area under the plasma homocysteine-time curve after methionine loading by 221 micro mol. 24 h/L (95% CI: -425, -16, P < 0.05) compared with placebo, whereas folic acid had no effect. In conclusion, betaine appears to be highly effective in preventing a rise in plasma homocysteine concentration after methionine intake in subjects with mildly elevated homocysteine. It is not known whether this potential of betaine to "stabilize" circulating homocysteine concentrations lowers the risk of cardiovascular disease.

2. Am J Clin Nutr. 2002 Nov;76(5):961-7.

Betaine supplementation decreases plasma homocysteine concentrations but does not affect body weight, body composition, or resting energy expenditure in human subjects.

Schwab U, Torronen A, Toppinen L, Alfthan G, Saarinen M, Aro A, Uusitupa M.

Department of Clinical Nutrition, University of Kuopio, Kuopio, Finland. ursula.schwab@uku.fi

BACKGROUND: Betaine (trimethylglycine) is found in several tissues in humans. It is involved in homocysteine metabolism as an alternative methyl donor and is used in the treatment of homocystinuria in humans. In pigs, betaine decreases the amount of adipose tissue. OBJECTIVE: The aim of the study was to examine the effect of betaine supplementation on body weight, body composition, plasma homocysteine concentrations, blood pressure, and serum total and lipoprotein lipids. DESIGN: Forty-two obese, white subjects (14 men, 28 women) treated with a hypoenergetic diet were randomly assigned to a betaine-supplemented group (6 g/d) or a control group given placebo for 12 wk. The intervention period was preceded by a 4-wk run-in period with a euenergetic diet. RESULTS: Body weight, resting energy expenditure, and fat mass decreased significantly in both groups with no significant difference between the groups. Plasma homocysteine concentrations decreased in the betaine group ( +/- SD: 8.76 +/- 1.63 micro mol/L at 4 wk, 7.93 +/- 1.52 micro mol/L at 16 wk; P = 0.030 for the interaction of time and treatment). Diastolic blood pressure decreased without a significant difference between the groups. Serum total and LDL-cholesterol concentrations were higher in the betaine group than in the control group (P < 0.05). CONCLUSION: A hypoenergetic diet with betaine supplementation (6 g daily for 12 wk) decreased the plasma homocysteine concentration but did not affect body composition more than a hypoenergetic diet without betaine supplementation did.

3. An Esp Pediatr. 2002 Apr;56(4):337-41.

[Neonatal onset methylmalonic aciduria and homocystinuria:Biochemical and clinical improvement with betaine therapy]

[Article in Spanish]

Urbon Artero A, Aldana Gomez J, Reig Del Moral C, Nieto Conde C, Merinero Cortes B.

Servicio de Pediatria, Hospital General de Segovia, Spain.

Methylmalonic aciduria and homocystinuria is a very rare inborn error of cellular cobalamin (Cbl) metabolism. We describe the biochemical evolution and clinical course of a boy with neonatal onset CblC mutant defect.Born after a normal pregnancy, the patient developed general hypotonia and severe feeding difficulties at 5 days of life. Diagnosis of methylmalonic aciduria and homocystinuria was established by amino-acid and organic acid analysis and was confirmed by enzyme and genetic studies.The patient was initially treated with parenteral hydroxocobalamin (1 mg/day), oral carnitine (100 mg/kg/day) and a restricted protein diet. This treatment returned methylmalonic acid levels to normal. Despite the parenteral hydroxocobalamin therapy, the patient showed no improvement in neurological dysfunction, hypotonia or developmental delay. Oral betaine supplementation (3 g/day) from months 3-15 reduced plasma total homocysteine and homocystinuria. The patient showed clinical improvement in neurological and growth development.We conclude that early betaine therapy was safe and effective in our patient with neonatal onset methylmalonic aciduria and homocystinuria type CblC.

4. Thromb Res. 2000 Jun 1;98(5):375-81.

Tissue factor pathway inhibitor levels in patients with homocystinuria.

Cella G, Burlina A, Sbarai A, Motta G, Girolami A, Berrettini M, Strauss W.

II Department of Medicine, University of Padua Medical School, Italy.

Thrombotic events are a well-recognized complication of homocystinuria. However, the mechanisms involved in the atherogenic and thrombotic effects of homocyst(e)ine remain incompletely understood. The objective of this study was to determine the role of endothelial cell activation/damage as indicated by levels of thrombomodulin, tissue factor and tissue factor pathway inhibitor, and factor VII activity in patients with homocystinuria. Six patients with homocystinuria, nonresponsive to pyridoxine, treated only with trimethylglycine (betaine) were injected with a bolus of 20 IU/kg body weight of unfractionated commercial heparin to induce the release of tissue factor pathway inhibitor from the vascular endothelium. Tissue factor, thrombomodulin, and factor VII activity were measured by enzyme-linked immunosorbent assay and clotting assay before heparin administration. Tissue factor pathway inhibitor antigen and activity were measured before and 5 minutes after the bolus of heparin. Levels of homocyst(e)ine were elevated (patients: 144.2+/-19.2 micromol/L; controls: 10.2+/-0.9 micromol/L); however, levels of thrombomodulin, tissue factor, and tissue factor pathway inhibitor antigen were not statistically different from the control group. In contrast, tissue factor pathway inhibitor activity showed a significantly increased level (patients: 2.09+/-0.34 U/L; controls: 1.14+/-0.20 U/L; p<0.05) that was correlated with homocyst(e)ine. Factor VII activity was significantly decreased (patients: 64.7+/-5.1%; controls: 91.4+/-4.7%; p<0.05) and inversely correlated with homocyst(e)ine. After heparin the patients released higher amounts of tissue factor pathway inhibitor antigen and activity compared with the control group; however, the difference was not statistically significant. Although not treated with antithrombotic drugs, none of the patients had any thromboembolic complications after starting betaine. In addition to betaine treatment, the enhanced factor pathway inhibitor antigen activity observed in this small series of patients suggests that factor pathway inhibitor antigen may play an additional, as yet unexplained, role in this genetic disorder.

5. J Inherit Metab Dis. 1997 Jun;20(2):295-300.

The natural history of vascular disease in homocystinuria and the effects of treatment.

Wilcken DE, Wilcken B.

Department of Cardiovascular Medicine, University of New South Wales, Australia.

Among 40 patients with homocystinuria due to cystathionine beta-synthase deficiency diagnosed in the state of New South Wales, Australia (population 6 million) and followed long-term, there were 10 deaths at ages 2-30 years. Of these 8 were definite vascular deaths, one was a presumed vascular death, and the other was due to an accident and unrelated to homocystinuria. The vascular deaths were all early cases and only one patient, a pyridoxine-responsive 30-year-old woman, had been prescribed adequate treatment although it was uncertain that she was taking it. In 32 patients of mean age 30 years (range 9-66 years) there were 539 patient-years of treatment with pyridoxine, folic acid and hydroxocobalamin. There were 17 pyridoxine-responsive patients and all maintained plasma total free homocyst(e)ine levels < 20 mumol/L over an average treatment period of 16.6 years. The 15 nonresponsive patients received additionally 6-9 g of betaine daily. This resulted in a further 74% mean decline (+/-14% SD) in plasma total free homocyst(e)ine, persisting during an average (post-betaine) treatment period of 11 years; current mean +/- SD levels are 33 +/- 17 mumol/L (n = 15). There were two vascular events during treatment, one fatal pulmonary embolus (see above) and one myocardial infarction, whereas without treatment, 21 would have been expected, chi2 = 14.22, p = 0.0001, relative risk 0.09 (95% CI 0.02-0.38). There were no events during 258 patient-years of treatment in the 15 pyridoxine-nonresponsive patients (p < 0.005 versus expected untreated). Nineteen patients had a total of 19 major and 15 minor operations requiring anaesthetic, and three had successful pregnancies, one whilst receiving betaine. There were no thromboembolic complications. We conclude that treatment which effectively lowers circulating homocyst(e)ine, even to suboptimal levels, markedly reduces cardiovascular risk in patients with cystathionine beta-synthase deficiency, and that betaine therapy contributes importantly to this in pyridoxine-nonresponsive patients. Betaine as additional therapy is safe and effective for at least 16 years.

6. Clin Chim Acta. 1991 Dec 31;204(1-3):239-49.

Betaine:homocysteine methyltransferase--a new assay for the liver enzyme and its absence from human skin fibroblasts and peripheral blood lymphocytes.

Wang JA, Dudman NP, Lynch J, Wilcken DE.

Department of Cardiovascular Medicine, Prince Henry Hospital, University of New South Wales, Sydney, Australia.

Chronic elevation of plasma homocysteine is associated with increased atherogenesis and thrombosis, and can be lowered by betaine (N,N,N-trimethylglycine) treatment which is thought to stimulate activity of the enzyme betaine:homocysteine methyltransferase. We have developed a new assay for this enzyme, in which the products of the enzyme-catalysed reaction between betaine and homocysteine are oxidised by performic acid before being separated and quantified by amino acid analysis. This assay confirmed that human liver contains abundant betaine:homocysteine methyltransferase (33.4 nmol/h/mg protein at 37 degrees C, pH 7.4). Chicken and lamb livers also contain the enzyme, with respective activities of 50.4 and 6.2 nmol/h/mg protein. However, phytohaemagglutinin-stimulated human peripheral blood lymphocytes and cultured human skin fibroblasts contained no detectable betaine:homocysteine methyltransferase (less than 1.4 nmol/h/mg protein), even after cells were pre-cultured in media designed to stimulate production of the enzyme. The results emphasize the importance of the liver in mediating the lowering of elevated circulating homocysteine by betaine.

7. Arch Dis Child. 1989 Jul;64(7):1061-4.

Betaine for treatment of homocystinuria caused by methylenetetrahydrofolate reductase deficiency.

Holme E, Kjellman B, Ronge E.

Department of Clinical Chemistry, Gothenburg University, Sweden.

A 24 day old girl with homocystinuria and hypomethioninaemia caused by methylenetetrahydrofolate reductase deficiency presented with rapidly progressing encephalopathy and myopathy. An almost complete recovery was achieved by treatment with betaine.

8. J Inherit Metab Dis. 1988;11(3):291-8.

The effect of oral betaine on vertebral body bone density in pyridoxine-non-responsive homocystinuria.

Gahl WA, Bernardini I, Chen S, Kurtz D, Horvath K.

Section of Human Biochemical Genetics, National Institute of Child Health and Human Development, Bethesda, Maryland 20892.

Five pyridoxine-non-responsive homocystinuric patients aged 5 to 32 years were treated with oral betaine, 3 g b.i.d, in a double-blind, placebo-controlled, two-year crossover study of its effect on bone mineralization. Betaine therapy significantly reduced mean plasma homocystine (36 +/- 9 (SEM) mumol L-1 to 9 +/- 4 mumol L-1), with variable increases in plasma methionine and no adverse effects. Bone density, measured by computerized tomographic scanning of vertebral bodies, was below normal in all patients at the start of the study, and was not significantly altered by betaine therapy administered according to this protocol.

9. Metabolism. 1985 Dec;34(12):1115-21.

Homocystinuria due to cystathionine beta-synthase deficiency--the effects of betaine treatment in pyridoxine-responsive patients.

Wilcken DE, Dudman NP, Tyrrell PA.

Homocystinuria due to cystathionine beta-synthase deficiency may be responsive to pyridoxine, a precursor of the cofactor pyridoxal phosphate, and the amount of residual enzyme activity present is the probable determinant of this. In six treated pyridoxine-responsive patients whose biochemical control of fasting plasma amino acid levels appeared optimal, we assessed the effects on plasma amino acids of standard oral methionine loads (4g/m2 of body area) before and after adding betaine (trimethylglycine) 6 g/d, to the treatment regimen of pyridoxine and folic acid. Our aim was to define the capacity of these patients to metabolize methionine and to determine whether betaine would effect a reduction in postload homocysteine levels. During the 24 hours after the methionine challenge all patients had higher plasma methionine and homocysteine and lower cysteine than did 17 normal subjects. After betaine these homocysteine responses were reduced to near normal, and there was a trend toward increased methionine. There was a direct correlation between premethionine fasting homocysteine and mean homocysteine responses during the 24 hours following the methionine load, both before (r = 0.79) and after betaine (r = 0.71). Betaine also increased plasma cysteine levels in patients with the more severe biochemical abnormalities. After betaine there were modest increases in plasma serine (mean increase 25%; P less than 0.025). Since the vascular complications of homocystinuria are related to increased plasma homocysteine, betaine therapy may reduce this risk in patients receiving a standard pyridoxine and folic acid regimen in whom there are abnormal homocysteine responses after a standard methionine load.

10. Eur J Pediatr. 1984 Jun;142(2):147-50.

Betaine in the treatment of homocystinuria due to 5,10-methylenetetrahydrofolate reductase deficiency.

Wendel U, Bremer HJ.

In a 3-year-old mentally retarded girl with homocystinuria due to 5,10-methylenetetrahydrofolate reductase deficiency among different therapeutic approaches only treatment with betaine (15-20 g/day) resulted in a satisfactory biochemical response. Betaine improved homocysteine remethylation and thus lowered plasma homocystine to trace amounts and normalized the previously very low plasma methionine concentration. This biochemical response was associated with a clinical improvement although she remained mentally retarded.

11. N Engl J Med. 1983 Aug 25;309(8):448-53.

Homocystinuria--the effects of betaine in the treatment of patients not responsive to pyridoxine.

Wilcken DE, Wilcken B, Dudman NP, Tyrrell PA.

The treatment of homocystinuria that is not responsive to pyridoxine is not usually biochemically or clinically successful, and vascular, ocular, and skeletal complications commonly supervene. Persistent marked homocysteinemia appears to be the most important biochemical disturbance leading to these complications. Ten patients with cystathionine beta-synthase deficiency that was not responsive to pyridoxine and one patient with homocystinuria due to a defect in cobalamin metabolism were treated with 6 g daily of betaine added to conventional therapy, to improve homocysteine remethylation. All patients had a substantial decrease in plasma total homocysteine levels (P less than 0.001) and an increase in total cysteine levels (P less than 0.001). Changes in plasma methionine concentrations were variable. Fasting levels of plasma amino acids became normal in two patients, and in six there was immediate clinical improvement. There were no unwanted effects. We conclude that treatment of homocystinuria that is not responsive to pyridoxine and of disorders of homocysteine remethylation should include betaine in adequate doses to ensure maximum lowering of elevated plasma homocysteine levels.

Dry mouth

12. Acta Odontol Scand. 1998 Apr;56(2):65-9.

Betaine-containing toothpaste relieves subjective symptoms of dry mouth.

Soderling E, Le Bell A, Kirstila V, Tenovuo J.

Institute of Dentistry, University of Turku, Finland.

Subjects with dry mouth often experience irritation of the oral mucosa when using sodium lauryl sulfate containing products for oral hygiene. Betaine, or trimethylglycine, reduces skin-irritating effects of ingredients of cosmetics such as sodium lauryl sulfate. The aim of the present study was to compare the effects of a betaine-containing toothpaste with a regular toothpaste on the oral microbial flora, the condition of the oral mucosa, and subjective symptoms of dry mouth in subjects with chronic dry mouth symptoms. Thirteen subjects with chronic dry mouth symptoms and with a paraffin-stimulated salivary flow rate < or = 1 mL/min participated in the double-blind crossover study. Ten subjects had a very low salivary flow rate (< or = 0.6 mL/min). The subjects used both experimental toothpastes (with or without 4% betaine) twice a day for 2 weeks. Oral examinations and microbiologic sample collections were made at the base lines preceding the two experimental periods and at the end. Standardized questions on subjective symptoms of dry mouth were used when the subjects were interviewed at the end of the two experimental periods. No study-induced significant changes were observed in the microbiologic variables (plaque index, mutans streptococci, lactobacilli, Candida species) or in the appearance of the oral mucosa. The use of the betaine-containing toothpaste was, however, associated with a significant relief of several subjective symptoms of dry mouth. Betaine appears thus to be a promising ingredient of toothpastes in general and especially of toothpastes designed for patients with dry mouth.

13. J Contemp Dent Pract. 2003 May 15;4(2):11-23.

Effects of a betaine-containing toothpaste on subjective symptoms of dry mouth: a randomized clinical trial.

Rantanen I, Tenovuo J, Pienihakkinen K, Soderling E.

Institute of Dentistry, University of Turku, Finland. irma.rantanen@utu.fi

Our aim was to study the effects of mildly flavoured sodium lauryl sulphate (SLS)-containing and detergent-free toothpastes with and without betaine (BET) on subjective symptoms of dry mouth in a randomised clinical trial. BET is an osmoprotectant that reacts with molecules to supply the surface with a water coating that protects cells from surfactants. Twenty-seven xerostomic patients and 18 healthy controls took part in the randomised, double-blind clinical trial with a crossover design. Three mildly flavoured toothpastes: (1) 4% BET, (2) 1% SLS and 4% BET, and (3) 1% SLS were used for six weeks each. The reference or washout paste contained neither SLS nor BET. The subjects' dental appointments were at the beginning of the trial and before and after the use of each toothpaste. At each appointment, the subjects were interviewed about subjective sensations of dry mouth (Visual Assessment Scoring (VAS) Index). The subjects did not report any adverse effects in connection with the use of the toothpastes. The VAS scores for lip dryness and eating difficulties were significantly lower for the BET paste (lip dryness: BET<BET+SLS; p < 0.005 and eating difficulties: BET<BET+SLS; p = 0.02; BET<reference; p = 0.003). The BET paste relieved dry mouth symptoms in 44% of the xerostomic patients, the corresponding figures for the other pastes being BET+SLS 22% (p = 0.002 as compared with BET), SLS 18% (p = 0.022), and reference 7% (p = 0.000). In conclusion, all the mildly flavoured toothpastes used in this study were well accepted by the xerostomic subjects. Thus, other toothpaste components may be more mucosa-irritating than just SLS, or else they enhance the effect of SLS. The detergent-free, BET-containing toothpaste appeared to be associated with relief of some symptoms of dry mouth.

Methylation

14. Life Sci. 1983 Feb 14;32(7):771-4.

Betaine, metabolic by-product or vital methylating agent?

Barak AJ, Tuma DJ.

The possible physiological role of betaine, the oxidative product of choline, is considered. It is proposed that betaine, instead of merely being a metabolic by-product of choline oxidation, may serve as an important methylating agent when normal methylating pathways are impaired by ethanol ingestion, drugs or nutritional imbalances. Furthermore, betaine may prove to have therapeutic application in cases of altered folate, vitamin B12 or methionine metabolism.

Affect on SAMe

15. Alcohol. 1996 Sep-Oct;13(5):483-6.

Betaine effects on hepatic methionine metabolism elicited by short-term ethanol feeding.

Barak AJ, Beckenhauer HC, Tuma DJ.

Liver Study Unit, VA Alcohol Research Center, Omaha, NE 68105, USA.

Previous studies in this laboratory have shown that feeding of ethanol to rats produces prompt inhibition of methionine synthetase (MS) as well as a subsequent increase in activity of betaine homocysteine methyltransferase (BHMT). Further studies have shown that supplemental dietary betaine enhanced methionine metabolism and S-adenosylmethionine (SAM) generation in control and ethanol-fed rats. Because MS and BHMT are both involved in the formation of SAM, this study was conducted to determine early effects of ethanol on hepatic SAM levels and the influence of betaine supplementation on parameters of methionine metabolism during the early periods of MS inhibition and enhanced BHMT activity. Results showed that ethanol feeding produced a significant loss in SAM in the first week with a return to normal SAM levels in the second week. Betaine feeding enhanced hepatic betaine pools in control as well as ethanol-fed animals. This feeding attenuated the early loss of SAM in ethanol-fed animals, produced an early increase in BHMT activity, and generated increased levels of SAM in both control and ethanol-fed groups. Furthermore, betaine lowered significantly the accumulation of hepatic triglyceride produced by ethanol after 2 weeks of ingestion.

16. Alcohol Clin Exp Res. 1993 Jun;17(3):552-5. (Animal Study)

Dietary betaine promotes generation of hepatic S-adenosylmethionine and protects the liver from ethanol-induced fatty infiltration.

Barak AJ, Beckenhauer HC, Junnila M, Tuma DJ.

Department of Veterans Affairs Medical Center, Omaha, Nebraska 68105.

Previous studies have shown that ethanol feeding to rats alters methionine metabolism by decreasing the activity of methionine synthetase. This is the enzyme that converts homocysteine in the presence of vitamin B12 and N5-methyltetrahydrofolate to methionine. The action of the ethanol results in an increase in the hepatic level of the substrate N5-methyltetrahydrofolate but as an adaptive mechanism, betaine homocysteine methyltransferase, is induced in order to maintain hepatic S-adenosylmethionine at normal levels. Continued ethanol feeding, beyond 2 months, however, produces depressed levels of hepatic S-adenosylmethionine. Because betaine homocysteine methyltransferase is induced in the livers of ethanol-fed rats, this study was conducted to determine what effect the feeding of betaine, a substrate of betaine homocysteine methyltransferase, has on methionine metabolism in control and ethanol-fed animals. Control and ethanol-fed rats were given both betaine-lacking and betaine-containing liquid diets for 4 weeks, and parameters of methionine metabolism were measured. These measurements demonstrated that betaine administration doubled the hepatic levels of S-adenosylmethionine in control animals and increased by 4-fold the levels of hepatic S-adenosylmethionine in the ethanol-fed rats. The ethanol-induced infiltration of triglycerides in the liver was also reduced by the feeding of betaine to the ethanol-fed animals. These results indicate that betaine administration has the capacity to elevate hepatic S-adenosylmethionine and to prevent the ethanol-induced fatty liver.

17. J Inherit Metab Dis. 1994;17(5):560-5.

Effect of betaine on S-adenosylmethionine levels in the cerebrospinal fluid in a patient with methylenetetrahydrofolate reductase deficiency and peripheral neuropathy.

Kishi T, Kawamura I, Harada Y, Eguchi T, Sakura N, Ueda K, Narisawa K, Rosenblatt DS.

Department of Pediatrics, Hiroshima University School of Medicine, Japan.

A 16-year-old Japanese girl with 5,10-methylenetetrahydrofolate reductase deficiency showed peripheral neuropathy. There were no significant responses to vitamin B6, vitamin B12 or folate, given alone or in combination. With the addition of betaine monohydrate, she has been free from gait disturbance and muscle weakness. The concentration of S-adenosylmethionine in cerebrospinal fluid, which was undetectable before receiving betaine monohydrate, increased to about the normal level 24 months after treatment with betaine monohydrate.

Alcoholic steatosis

18. Alcohol Clin Exp Res. 1997 Sep;21(6):1100-2.

The effect of betaine in reversing alcoholic steatosis.

Barak AJ, Beckenhauer HC, Badakhsh S, Tuma DJ.

Veterans Affairs Alcohol Research Center, Department of Veterans Affairs Medical Center, Omaha, Nebraska 68105, USA.

The feeding of ethanol to experimental animals results in fatty infiltration of the liver. Recent findings have shown that ethanol-induced steatosis is accompanied by a lowering in hepatic S-adenosylmethionine (SAM) levels. It is known that SAM provides substrates for reduced glutathione formation and offers the cell protection from toxic metabolic oxidants. A recent study in this laboratory demonstrated that dietary supplementation with betaine generated increased SAM in the liver and protected against ethanol-induced steatosis. The present study not only showed that betaine supplementation to rats protects the liver from alcoholic steatosis, but also demonstrated that once steatosis is established, treatment with betaine partially reversed the steatosis after cessation of ethanol feeding. Furthermore, this study indicated that betaine supplementation to the diet had the capacity to attenuate steatosis despite the continued feeding of ethanol.

Concentrations

19. Clin Chim Acta. 1988 Dec 30;178(3):241-9. (Animal Study)

Betaine metabolism in human neonates and developing rats.

Davies SE, Chalmers RA, Randall EW, Iles RA.

Medical Unit (Section of Metabolism and Endocrinology), London Hospital Medical College, Whitechapel.

Proton nuclear magnetic resonance spectroscopy has been used to demonstrate the presence of high concentrations of betaine (up to 0.75 mol/mol creatinine) in the urine of normal healthy human neonates. Betaine is not normally excreted in adults. Excretion of betaine from birth to 7 days old was monitored. The excretion of betaine in rats from 21 days after birth to 40-45 days old was also monitored. A peak in excretion in the rats of 1.5-3 mol/mol creatinine occurred between days 30-35. The presence of a high concentration of betaine in the urine is unlikely to be caused by a relative lack of betaine homocysteine methyl transferase activity compared with adults but may relate to the disposal of dietary choline during development.

Liver support

20. Vet Pathol. 2000 May;37(3):231-8. (Animal Study)

Reduction of carbon tetrachloride-induced hepatotoxic effects by oral administration of betaine in male Han-Wistar rats: a morphometric histological study.

Junnila M, Rahko T, Sukura A, Lindberg LA.

Department of Basic Veterinary Sciences: Veterinary Pathology, University of Helsinki, Faculty of Veterinary Medicine, Finland. matti.junnila@pp.inet.fi

Eighty-five male Han-Wistar rats were arranged into three groups: CCl4-exposed rats, CCl4 + betaine-exposed rats, and control rats. To see the effect of betaine alone, five rats of the control and of the CCl4 + betaine groups were sacrificed after 7 days, before exposure to CCl4. After that, two of the groups (the CCl4 and CCl4 + betaine groups) were exposed to CCl4 (1 ml/kg per day subcutaneously [SC] for 4 consecutive days), and one of the groups (control group) was given olive oil (1 ml/kg per day SC for 4 consecutive days). At the start of the study (day 0), day 1, day 2, day 3, day 4, and 3 days after the last CCl4 and olive oil injections (day 7), samples of five rats per group were sacrificed, and the livers were taken for chemical analyses and histological examination. Oral betaine, after the acclimation period of a week, increased the number of mitochondria but not mitochondria size (day 0), compared with the case in control rats. Exposure to CCl4 resulted in centrilobular hepatic steatosis, and the administration of betaine significantly reduced this. Morphometric analyses also revealed that the addition of betaine increased the volume density of rough endoplasmic reticulum (RER) in the perinuclear areas of liver cell cytoplasm (day 7). Additionally, the administration of betaine prevented the reduction of Golgi complexes and mitochondrial figures in the cytoplasm observed after the exposures to CCl4. Also, the volume density of mitochondria was smallest in the CCl4-group, but the difference was not statistically significant. The results indicate that oral betaine either improves recovery or reduces the toxic effects of CCl4 on cell organelles in liver cells of male Han-Wistar rats.

21. Vet Hum Toxicol. 1998 Oct;40(5):263-6. (Animal Study)

Betaine reduces hepatic lipidosis induced by carbon tetrachloride in Sprague-Dawley rats.

Junnila M, Barak AJ, Beckenhauer HC, Rahko T.

University of Helsinki, Faculty of Veterinary Medicine, Department of Basic Veterinary Sciences-Veterinary Pathology, Finland.

Carbon tetrachloride-injected rats were given liquid diets with and without betaine for 7 d. Hepatic lipidosis was induced by 4 daily injections of carbon tetrachloride (CCl4). Animals were killed and their livers and blood taken for analysis of betaine, S-adenosylmethionine (SAM), betaine homocysteine methyltransferase (BHMT), triglyceride, alanine aminotransferase and aspartate aminotransferase. Liver samples were also processed and stained for histological examination. Supplemental betaine reduced triglyceride in the liver and centrilobular hepatic lipidosis induced by the CCl4 injections. In both the control and experimental groups receiving betaine, liver betaine, BHMT and SAM were significantly higher than in their respective groups not receiving betaine. This study provides evidence that betaine protects the liver against CCl4-induced lipidosis and may be a useful therapeutic and prophylactic agent in ameliorating the harmful effects of CCl4.

22. Hepatology. 1998 Mar;27(3):787-93. (Animal Study)

Betaine as an osmolyte in rat liver: metabolism and cell-to-cell interactions.

Wettstein M, Weik C, Holneicher C, Haussinger D.

Clinic for Gastroenterology, Hepatology, and Infectiology, Heinrich-Heine-University, Dusseldorf, Germany.

Betaine was recently identified as an osmolyte in rat liver macrophages (Kupffer cells [KCs]) and sinusoidal endothelial cells (SECs). Betaine interferes with KC functions, such as phagocytosis, cytokine, and prostaglandin syntheses. As betaine is derived from choline, the present study was undertaken to evaluate osmosensitivity and cell heterogeneity of choline metabolism in rat liver. In the perfused rat liver after in vivo prelabeling with [14C]-choline, hypoosmotic stress induced a radioactivity release into the perfusate which was identified as [14C]-betaine by high-performance liquid chromatography (HPLC) analysis and which was inhibited by the anion exchanger inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid. Choline metabolism was studied in cultured liver parenchymal cells, (PCs), KCs, and SECs. Choline was taken up by all but betaine formation from choline was only detectable in PCs and not in KCs and SECs. Betaine formation in PCs was not stimulated by hyperosmolarity; rather, betaine has a role as an osmolyte in KCs and SECs but is of minor importance in PCs, as evidenced by only minor hyperosmolarity-induced betaine uptake. Thus, liver PCs can produce and release betaine derived from choline, and, thereby, possibly supply the osmolyte important for KC and SEC cell function. This may be another example for cell-to-cell interaction in the liver.

23. Hepatology. 1997 Dec;26(6):1560-6. (Animal Study)

Cytoprotection by the osmolytes betaine and taurine in ischemia-reoxygenation injury in the perfused rat liver.

Wettstein M, Haussinger D.

Clinic for Gastroenterology, Hepatology, and Infectiology, Heinrich-Heine-University, Dusseldorf, Germany.

Medium osmolarity sensitively regulates Kupffer cell functions like phagocytosis and prostaglandin (PG) and cytokine production. Betaine and taurine, recently identified as osmolytes in liver cells, interfere with these effects. Because Kupffer cell activation is an important pathogenic mechanism in ischemia-reoxygenation injury, the influence of osmolarity and osmolytes was investigated in a rat liver perfusion model of warm ischemia. Livers were perfused with different medium osmolarities for 60 to 90 minutes in the absence of oxygen, followed by another 90 minutes of reoxygenation. Lactate dehydrogenase (LDH) leakage into the effluent perfusate during the hypoxic and the reoxygenation period was eight- to 10-fold higher with a medium osmolarity of 385 mosmol/L than in normo-osmolarity, and further decreased with hypo-osmolar perfusion buffer. Betaine and taurine addition to the perfusate in near physiological concentrations decreased hypoxia-reoxygenation-induced LDH leakage, aspartate transaminase (AST) leakage, and perfusion pressure increase in hyperosmolar and normo-osmolar perfusions. Stimulation of PGD2, PGE2, thromboxane B2 (TXB2), and tumor necrosis factor alpha (TNF-alpha) release, as well as induction of carbon uptake by the liver during reoxygenation, were suppressed by betaine and taurine, pointing to an interference of these osmolytes with Kupffer cell function. In contrast, endothelial cell function as assessed by hyaluronic acid (HA) uptake was not influenced. It is concluded that warm ischemia-reoxygenation injury in rat liver is aggravated by hyperosmolarity and attenuated by hypo-osmolarity. The osmolytes betaine and taurine have a protective effect, presumably by inhibition of Kupffer cell activation.

Cryoprotective agent

24. Zentralbl Veterinarmed A. 1989 Feb;36(2):110-4. (Animal Study)

A preliminary study on the use of betaine as a cryoprotective agent in deep freezing of stallion semen.

Koskinen E, Junnila M, Katila T, Soini H.

In a preliminary experiment, betaine was added in concentrations ranging from 0 to 3.0 percent to stallion semen diluted with 4% glycerol extender. Motility of frozen-thawed semen was better in the extenders with high betaine concentration than in those of low concentration or the control. In a subsequent experiment, betaine was added to extended semen from five stallions to make a 2.5% betaine concentration. Two different cooling rates were used. The effect of betaine on spermatozoal motility was positive at both cooling rates studied. Motility of frozen-thawed 2.5% betaine semen samples was significantly (p less than 0.001) higher than that of the control samples.

Irritating properties of detergents

25. Skin Res Technol. 2003 Feb;9(1):50-8.

The ability of betaine to reduce the irritating effects of detergents assessed visually, histologically and by bioengineering methods.

Nicander I, Rantanen I, Rozell BL, Soderling E, Ollmar S.

Department of Dermatology I 43, Huddinge University Hospital, SE-14186 Huddinge, Sweden. ingrid.nicander@cob.ke.se

BACKGROUND/AIMS: A novel approach for reducing the undesired irritating properties of detergents on skin might be offered by betaine, which is a natural product derived from the sugar beet. The aim of the study was to explore the ability of betaine to reduce the irritating effects of two surfactants, sodium lauryl sulphate (SLS) and cocoamidopropylbetaine (CAPB). For evaluation of changes in skin reactions visual scoring, electrical impedance, transepidermal water loss and histology were used. METHODS: Twenty-one healthy subjects were patch tested for 24 h with SLS and CAPB alone and together with betaine, betaine alone, and the two controls distilled water and an unoccluded test site on both volar forearms. Responses were evaluated by measuring electrical impedance and transepidermal water loss before exposure and 24 h after the removal of the test substances, and also by visual inspection and histology. The electrical impedance device enables measurements at 31 frequencies and relevant information was extracted from the spectra using four indices. RESULTS: CAPB was found to be less irritating than SLS. The used detergents gave rise to distinctive impedance patterns also reflected by different types of histopathological skin responses. After the adding of betaine, the irritant reaction decreased for both detergents. CONCLUSIONS: Betaine is a promising ingredient to reduce the side effects of detergents and electrical impedance is a suitable tool both to quantify the degree of irritation as well as to differentiate between various types of reactions. Copyright Blackwell Munksgaard 2003

Pharmacokinetics

26. Br J Clin Pharmacol. 2003 Jan;55(1):6-13.

Pharmacokinetics of oral betaine in healthy subjects and patients with homocystinuria.

Schwahn BC, Hafner D, Hohlfeld T, Balkenhol N, Laryea MD, Wendel U.

Department of Paediatrics, Medical Faculty, Heinrich-Heine-University, Moorenstrasse 5, D-40225 Dusseldorf, Germany. schwahn@med.uni-duesseldorf.de

AIMS: Large oral doses of betaine have proved effective in lowering plasma homocysteine in severe hyperhomocysteinaemia. The pharmacokinetic characteristics and metabolism of betaine in humans have not been assessed and drug monitoring for betaine therapy is not available. We studied the pharmacokinetics of betaine and its metabolite dimethylglycine (DMG) in healthy subjects and in three patients with homocystinuria. METHODS: Twelve male volunteers underwent an open-label study. After one single administration of 50 mg betaine kg-1 body weight and during continuous intake of twice daily 50 mg kg-1 body weight, serial blood samples and 24 h urines were collected to determine betaine and DMG plasma concentrations and urinary excretion, respectively. Patients were evaluated after one single dose of betaine. RESULTS: We found rapid absorption (t(1/2),abs 00.28 h, s.d. 0.17) and distribution (t(1/2), lambda1 00.59 h, s.d. 0.22) of betaine. A Cmax of 0.94 mmol l-1 (s.d. 0.19) was reached after tmax 00.90 h (s.d. 0.33). The elimination half life t(1/2), z was 14.38 h (s.d. 7.17). After repeated dosage, t(1/2), lambda1 (01.77 h, s.d. 0.75) and t(1/2), z (41.17 h, s.d. 13.50) increased significantly (95% CI 0.73, 01.64 h and 19.90, 33.70 h, respectively), whereas absorption remained unchanged. DMG concentrations increased significantly after betaine administration and accumulation occurred to the same extent as with betaine. Renal clearance was low and urinary excretion of betaine was equivalent to 4% of the ingested dose. Distribution and elimination kinetics in homocystinuric patients appeared to be accelerated. CONCLUSIONS: Betaine plasma concentrations change rapidly after ingestion. Elimination half-life increased during continuous dosing over 5 days. Betaine is mainly eliminated by metabolism. More pharmacokinetic and pharmacodynamic studies in hyperhomocysteinaemic patients are needed to refine the current treatment with betaine.

27. Br J Clin Pharmacol. 2002 Aug;54(2):140-6.

An indirect response model of homocysteine suppression by betaine: optimising the dosage regimen of betaine in homocystinuria.

Matthews A, Johnson TN, Rostami-Hodjegan A, Chakrapani A, Wraith JE, Moat SJ, Bonham JR, Tucker GT.

Department of Chemical Pathology & Neonatal Screening, Sheffield Children's Hospital, Manchester, UK.

AIMS: To investigate the pharmacokinetics (PK) and pharmacodynamics (PD) of betaine in the treatment of classical homocystinuria due to cystathionine beta-synthase (CbetaS) deficiency with a view to optimizing the dosage regimen. METHODS: Betaine was given as a single oral dose of 100 mg kg(-1) to six patients (age range 6-17 years) who normally received betaine but whose treatment had been suspended for 1 week prior to the study. Plasma betaine and total homocysteine concentrations were measured by high performance liquid chromatography (h.p.l.c.) at frequent intervals over 24 h. The best-fit PK model was determined using the PK-PD program Win-Nonlin and the concentration-time-effect data analysed by an indirect PD model. Using the PK and PD parameters, simulations were carried out with the aim of optimizing betaine dosage. RESULTS: Betaine PK was described by both mono- and bi-exponential disposition functions with first order absorption and a lag time. The correlation coefficient between betaine oral clearance and body weight was 0.6. Mean betaine clearance was higher in males than in females (P=0.03). PK-PD simulation indicated minimal benefit from exceeding a twice-daily dosing schedule and a 150 mg kg(-1) day(-1) dosage for betaine. CONCLUSIONS: PK-PD modelling allows recommendations for optimal dosage of betaine in the treatment of homocystinuria, that have the potential for improved patient compliance and both therapeutic and pharmacoeconomic benefit.

Renal failure

28. Kidney Int. 2002 Mar;61(3):1040-6.

Betaine supplementation decreases post-methionine hyperhomocysteinemia in chronic renal failure.

McGregor DO, Dellow WJ, Robson RA, Lever M, George PM, Chambers ST.

Department of Nephrology, Christchurch Hospital and School of Medicine, Private Bag 4710, Christchurch, New Zealand. david.mcgregor@cdhb.govt.nz

BACKGROUND: Fasting and post-methionine load hyperhomocysteinemia are independent risk factors for vascular disease that are common in chronic renal failure. Folate decreases but seldom normalizes fasting total homocysteine (tHcy) concentrations in such patients. Glycine betaine (GB) is known to decrease tHcy in other clinical settings, but whether it is beneficial in chronic renal failure has not been established. METHODS: We conducted a crossover-controlled trial in 36 patients with chronic renal failure to determine if oral GB decreased fasting or post-methionine tHcy concentrations. All subjects received, in randomized sequence, 5-mg folic acid and 50-mg pyridoxine daily, with or without GB 4-g daily, for three months each. Fasting plasma tHcy, GB, folate, B vitamins, serum lipids and creatinine were measured at one and three months, and methionine load tests were performed at the end of each three-month treatment phase. RESULTS: GB and N,N-dimethylglycine (DMG) levels in plasma and urine increased markedly during GB treatment. Fasting tHcy decreased from baseline with both treatments but did not differ between treatments. Post-methionine tHcy decreased with both treatments and was 18% lower on GB than on folate and pyridoxine alone (P < 0.001). There were small increases in lipids during treatment with GB but the ratio of total: HDL cholesterol was unchanged. CONCLUSIONS: GB supplementation had no effect on fasting tHcy in patients with chronic renal failure who were folate and pyridoxine replete, but it significantly decreased tHcy concentrations after methionine loading.

Nonalcoholic steatohepatitis

29. Am J Gastroenterol. 2001 Sep;96(9):2711-7.

Comment in: Am J Gastroenterol. 2001 Sep;96(9):2534-6.

Betaine, a promising new agent for patients with nonalcoholic steatohepatitis: results of a pilot study.

Abdelmalek MF, Angulo P, Jorgensen RA, Sylvestre PB, Lindor KD.

Divisions of Gastroenterology and Hepatology and Surgical Pathology, Mayo Clinic and Foundation, Rochester, Minnesota 55905, USA.

OBJECTIVES: No effective therapy currently exists for patients with nonalcoholic steatohepatitis (NASH). Betaine, a naturally occurring metabolite of choline, has been shown to raise S-adenosylmethionine (SAM) levels that may in turn play a role in decreasing hepatic steatosis. Our aim was to determine the safety and effects of betaine on liver biochemistries and histological markers of disease activity in patients with NASH. METHODS: Ten adult patients with NASH were enrolled. Patients received betaine anhydrous for oral solution (Cystadane) in two divided doses daily for 12 months. Seven out of 10 patients completed 1 yr of treatment with betaine. RESULTS: A significant improvement in serum levels of aspartate aminotransferase (p = 0.02) and ALAT (p = 0.007) occurred during treatment. Aminotransferases normalized in three of seven patients, decreased by >50% in three of seven patients, and remained unchanged in one patient when compared to baseline values. A marked improvement in serum levels of aminotransferases (ALT -39%; AST -38%) also occurred during treatment in those patients who did not complete 1 yr of treatment. Similarly, a marked improvement in the degree of steatosis, necroinflammatory grade, and stage of fibrosis was noted at 1 yr of treatment with betaine. Transitory GI adverse events that did not require any dose reduction or discontinuation of betaine occurred in four patients. CONCLUSIONS: Betaine is a safe and well tolerated drug that leads to a significant biochemical and histological improvement in patients with NASH. This novel agent deserves further evaluation in a randomized, placebo-controlled trial.

30. Arzneimittelforschung. 2000 Aug;50(8):722-7.

Efficacy and safety of oral betaine glucuronate in non-alcoholic steatohepatitis. A double-blind, randomized, parallel-group, placebo-controlled prospective clinical study.

Miglio F, Rovati LC, Santoro A, Setnikar I.

International Drug Division, Hospital S. Orsola-Malpighi, Bologna, Italy.

In a prospective, randomized, double-blind therapeutic trial, 191 patients with non-alcoholic steatohepatitis were treated for 8 weeks daily b.i.d. orally either with betaine glucuronate combined with diethanolamine glucuronate and nicotinamide ascorbate (Ietepar) (96 patients) or with undistinguishable placebo capsules (95 patients). The verum treatment effectively reduced by 25% hepatic steatosis (p < 0.01) and by 6% hepatomegaly (p < 0.05), while placebo did not significantly reduce the disorders. Verum was also more effective than placebo on discomfort in abdominal upper right quadrant. The global efficacy of treatment was rated by the doctor "very good" or "good" in 48% of verum treated patients and only in 17% after placcbo (P of difference = 9 x 10(-6)). 52% of patients self-rated efficacy as "very good" or "good" after verum and only 34% after placebo (P of difference = 0.017). The verum treatment provoked a significant reduction of the increased liver transaminases (ALT, AST and gamma-GT) while placebo was ineffective. Adverse events were recorded in 10% of verum-treated patients and in 7% under placebo (no significant difference). In both groups the adverse events were mild and transient, did not require treatment discontinuation and were undistinguishable from common symptoms of liver disorders. In conclusion, the 8-week treatment with betaine glucuronate combined with diethanolamine glucuronate and nicotinamide ascorbate was found effective in non-alcoholic steatohepatitis, a disorder for which the hitherto pharmacological interventions were poorly and inconsistently effective.

Atherosclerosis

31. Biull Eksp Biol Med. 1987 Jul;104(7):30-2. (Animal Study)

[Cholagogic effect of trimethylglycine in normal animals of different ages and in experimental atherosclerosis]

[Article in Russian]

Zapadniuk VI, Panteleimonova TN.

Trimethylglycine at a dose of 1.5 g/kg was found to produce marked bile secretory effect in young and old rats. In rabbits with experimental atherosclerosis, trimethylglycine increased the content of biliary acids in the bile and normalized the indexes of lipid metabolism in the blood serum. Apparently, the effect on cholesterol transformation into biliary acids and its excretion with the bile is one of the mechanisms of anti-atherosclerotic action of trimethylglycine.

32. Farmakol Toksikol. 1986 Jul-Aug;49(4):71-3. (Animal Study)

[Corrective effect of trimethylglycine on the nicotinamide coenzyme and adenine nucleotide content of the tissues in experimental atherosclerosis]

[Article in Russian]

Zapadniuk VI, Chekman IS, Panteleimonova TN, Tumanov VA.

Experiments on adult rabbits with experimental atherosclerosis induced by cholesterol (0.25 g/kg for 90 days) showed that chronic administration of trimethylglycine (1.5 g/kg for 30 days) prevented a decrease of the liver and myocardium content of nicotinamide coenzymes and adenine nucleotides.

33. Farmakol Toksikol. 1983 Jul-Aug;46(4):83-5. (Animal Study)

[Effect of trimethylglycine on lipid metabolism in experimental atherosclerosis in rabbits]

[Article in Russian]

Panteleimonova TN, Zapadniuk VI.

It has been shown in adult rabbits aged 8 months with experimental cholesterol atherosclerosis that administration of trimethylglycinee in a dose of 0.5 g/kg reduces the elevated content of total and ester-bound cholesterol, beta-lipoproteins, total lipids in the blood serum and that of total cholesterol and triglycerides in the liver. Little toxicity and high efficacy of trimethylglycin in experimental atherosclerosis make this compound prospective in the light of its use as an antisclerotic agent.

Anticonvulsant activity

34. Pharmacol Biochem Behav. 1985 Apr;22(4):641-3.

Prevention of strychnine-induced seizures and death by the N-methylated glycine derivatives betaine, dimethylglycine and sarcosine.

Freed WJ.

Betaine (N,N,N-trimethylglycine) and N,N-dimethylglycine have been reported to have anticonvulsant properties in animals. The purpose of the present study was to determine whether these compounds can antagonize strychnine-induced seizures when administered intraperitoneally and to compare their effects with those of sarcosine (N-methylglycine) and glycine. Betaine, N,N-dimethylglycine and sarcosine were equipotent in decreasing the incidence of seizures and death, causing a 38 to 72 percent decrease in the incidence of seizures and death at a dosage of 5 mmole/kg. Glycine had no effect. Thus anticonvulsant activity is conferred to glycine by a single N-methylation.

Antimicrobial activity

35. Antimicrob Agents Chemother. 1990 Oct;34(10):1949-54.

Antimicrobial activity of betaine esters, quaternary ammonium amphiphiles which spontaneously hydrolyze into nontoxic components.

Lindstedt M, Allenmark S, Thompson RA, Edebo L.

Department of Clinical Bacteriology, University of Goteborg, Sweden.

A series of quaternary ammonium compounds that are esters of betaine and fatty alcohols with hydrocarbon chain lengths of 10 to 18 carbon atoms were tested with respect to antimicrobial activities and rates of hydrolysis. When the tetradecyl derivative was tested against some selected microorganisms, the killing effect was comparable to that of the stable quaternary ammonium compound cetyltrimethylammonium bromide. At higher pH values, both the antimicrobial effect and the rate of hydrolysis of the esters increased. However, whereas at pH 6 greater than 99.99% killing of Salmonella typhimurium was achieved with 5 micrograms/ml in 3 min, the rate of hydrolysis was less than 20% in 18 h. At pH 7, a similar killing effect was achieved in 2 min and 50% hydrolysis occurred in ca. 5 h. Thus, it is possible to exploit the rapid microbicidal effect of the compounds before they hydrolyze. The rate of hydrolysis was reduced by the presence of salt. The bactericidal effect of the betaine esters increased with the length of the hydrocarbon chain of the fatty alcohol moiety up to 18 carbon atoms. Since the hydrolysis products are normal human metabolites, the hydrolysis property may extend the use of these quaternary ammonium compounds as disinfectants and antiseptics for food and body surfaces.