Betaine Treatment Research Articles

High-fat diet plus carbon tetrachloride-induced liver fibrosis is alleviated by betaine treatment in rats

Steatosis, the first lesion in non-alcoholic fatty liver disease (NAFLD), may progress to fibrosis, cirrhosis, and hepatocellular carcinoma. Steatosis predisposes the liver to oxidative stress, inflammation, and cytokines. Betaine (BET) has antioxidant, antiinflammatory and hepatoprotective effects. However, the effects of BET on liver fibrosis development are unknown. Rats were treated with high-fat diet (60% of total calories from fat) for 14weeks. Carbon tetrachloride (0.2mL/kg; two times per week; i.p.) was administered to rats in the last 6weeks with/without commercial food containing BET (2%; w/w). Serum liver function tests and tumor necrosis factor-α, insulin resistance, hepatic triglyceride (TG) and hydroxyproline (HYP) levels and oxidative stress parameters were determined along with histopathologic observations. Alpha-smooth muscle-actin (α-SMA), transforming growth factor-β1 (TGF-β1) and type I collagen (COL1A1) protein expressions and mRNA expressions of matrix metalloproteinase-2 (MMP-2) and its inhibitors (TIMP-1 and TIMP-2) were evaluated. BET decreased TG and HYP levels, prooxidant status and fibrotic changes in the liver. α-SMA, COL1A1 and TGF-β1 protein expressions, MMP-2, TIMP-1, and TIMP-2 mRNA expressions diminished due to BET treatment. BET has an antifibrotic effect and this effect may be related to its antioxidant and antiinflammatory actions together with suppression on HSC activation.

Alleviation of hepatic fat accumulation by betaine involves reduction of homocysteine via up-regulation of betaine-homocysteine methyltransferase (BHMT)

We investigated the anti-lipogenic effect of betaine in rats fed methionine and choline-deficient diet (MCD). Intake of MCD for 3 wk resulted in a significant accumulation of hepatic lipids, which was prevented by betaine supplementation in drinking water (1%). Phosphorylation of AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), sterol regulatory element-binding protein-1c (SREBP-1c), and liver kinase B1 (LKB1) was inhibited by MCD intake, and these changes were all inhibited by betaine feeding. Meanwhile, betaine supplementation reversed the reduction of methionine and S-adenosylmethionine (SAM), and the elevation of homocysteine levels in the liver, which could be attributable to the induction of betaine-homocysteine methyltransferase (BHMT) and methionine adenosyltransferase (MAT). Different cell lines were used to clarify the role of homocysteine on activation of the AMPK pathway. Homocysteine treatment decreased pAMPK, pACC, pSREBP-1c and pLKB1 in HepG2 cells. Metformin-induced activation of AMPK was also inhibited by homocysteine. Treatment with hydroxylamine, a cystathionine β-synthase inhibitor, resulted in a reduction of pAMPK, pACC and pSREBP-1c, accompanied by an elevation of intracellular homocysteine. Betaine treatment prevented the homocysteine-induced reduction of pAMPK, pACC, pSREBP-1c and pLKB1 in H4IIE cells, but not in HepG2 cells. Also the elevation of cellular homocysteine and inhibition of protein expression of BHMT were prevented by betaine only in H4IIE cells which express BHMT. The results suggest that the beneficial effect of betaine against hepatic lipid accumulation may be attributed, at least in part, to the depletion of homocysteine via up-regulation of BHMT in hepatocytes.

Betaine treatment decreased oxidative stress, inflammation, and stellate cell activation in rats with alcoholic liver fibrosis.

The aim of this study was to investigate the effect of betaine (BET) on alcoholic liver fibrosis in rats. Fibrosis was experimentally generated with ethanol plus carbon tetrachloride (ETH+CCl4) treatment. Rats were treated with ETH (5% v/v in drinking water) for 14 weeks. CCl4 was administered intraperitoneally (i.p.) 0.2mL/kg twice a week to rats in the last 6 weeks with/without commercial food containing BET (2% w/w). Serum hepatic damage markers, tumor necrosis factor-α, hepatic triglyceride (TG) and hydroxyproline (HYP) levels, and oxidative stress parameters were measured together with histopathologic observations. In addition, α-smooth muscle-actin (α-SMA), transforming growth factor-β1 (TGF-β1) and type I collagen (COL1A1) protein expressions were assayed immunohistochemically to evaluate stellate cell (HSC) activation. mRNA expressions of matrix metalloproteinase-2 (MMP-2) and its inhibitors (TIMP-1 and TIMP-2) were also determined. BET treatment diminished TG and HYP levels; prooxidant status and fibrotic changes; α-SMA, COL1A1 and TGF-β protein expressions; MMP-2, TIMP-1 and TIMP-2 mRNA expressions in the liver of fibrotic rats. In conclusion, these results indicate that the antifibrotic effect of BET may be related to its suppressive effects on oxidant and inflammatory processes together with HSC activation in alcoholic liver fibrosis.

Effect of hot water combined with glycine betaine alleviates chilling injury in cold-stored loquat fruit

The effect of hot water dipping (HW, 45°C for 10min) combined with glycine betaine (GB, 10mmolL−1) treatment on chilling injury (CI), fruit quality and physiological changes in loquat fruit stored at 1°C were investigated. The results showed that treatment with HW or GB alone both significantly alleviated CI in loquat fruit compared with the control. The combined treatment of HW and GB was more effective in reducing CI and maintaining quality parameters than HW or GB alone. In addition, HW combined with GB treatment significantly reduced ion leakage, and malondialdehyde (MDA) content in loquat fruit. The activities of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) were higher in the combined treatment than those in the control. The accumulation of proline and γ-aminobutyric acid (GABA) contents in loquat fruit were also induced by the combination of HW and GB, which was explained by increase in metabolism enzyme activities such as Δ1-pyrroline-5-carboxylate synthetase (P5CS), ornithine δ-aminotransferase (OAT) and glutamate decarboxylase (GAD). These results suggest that HW combined with GB treatment enhanced chilling tolerance in loquat fruit by inducing antioxidant enzymes activities and proline or GABA metabolism. The combination of HW and GB treatment could be a useful method to reduce CI in cold-stored loquat fruit, which has potential for application in the loquat industry.

Effect of natural betaine on estimates of semen quality in mature AI boars during summer heat stress

This study evaluated the effect of supplemental dietary betaine at three concentrations (0.0%, 0.63% and 1.26%) on semen characteristics, quality and quality after storage on boars. The trial was conducted between 22 July and 1 October 2014 in a boar stud located in Oklahoma. Boars were blocked by age within genetic line and randomly allotted to receive 0% (CON, n (line T)=22, n (line L)=10), 0.63% (BET-0.63%, n (line T)=21, n (line L)=6) or 1.26% (BET-1.26%, n (line T)=23, n (line L)=7). The diets containing betaine were fed over 10 weeks, to ensure supplemental betaine product (96% betaine) daily intakes of 16.34 and 32.68g, for the BET-0.63% and BET-1.26% diets, respectively. Serum homocysteine concentrations were less for animals with betaine treatments (P=0.016). Rectal temperatures of the boars were unaffected by betaine diets. Betaine tended to increase total sperm in the ejaculates when collectively compared with data of the control animals (P=0.093). Sperm morphology analysis indicated there was a greater percent of sperm with distal midpiece reflex (P=0.009) and tail (P=0.035) abnormalities in boars fed the BET-1.26% than boars fed the BET-0.63% diet. Betaine concentration in the seminal plasma was greater in boars with betaine treatments, with animals being fed the 0.63% and 1.26% diets having 59.2% and 54.5% greater betaine concentrations in seminal plasma as compared with boars of the control group (P=0.046). In conclusion, betaine supplementation at 0.63% and 1.26% tended to increase sperm concentration in the ejaculates by 6% and 13%, respectively, with no negative impacts on semen quality when 0.63% of betaine was included in the diet.

Chemically enhanced lipid production from microalgae under low sub-optimal temperature

Low sub-optimal temperatures are one of the major environmental factors that significantly affects growth and lipid production of microalgae. However, until now little research has been done to estimate and reduce the effects of low sub-optimal temperatures (typically 15°C–20°C) on productivity of algal lipids, which serve as feedstock for biodiesel. We addressed this issue by growing the microalga Chlorella sorokiniana under a low sub-optimal temperature (18°C) (the optimal temperature was 26°C, which was used as a control). Algal growth was inhibited at 18°C compared to 26°C, resulting in overall decreased lipid productivity; however, decreased temperature increased the total lipid content of the algal cell. Importantly, decreased lipid productivity under the low sub-optimal temperature was improved by an application of exogenous glycine betaine at 500mg/L compared to the control treatment of 26°C. Quantitative PCR analysis showed that exogenous glycine betaine enhanced expression of genes encoding Rubisco, one of the key enzymes involved in carbon fixation through photosynthesis. The expression of other key genes involved in the Calvin cycle was up-regulated by treatment with glycine betaine at both 18°C and 26°C. In addition, glycine betaine treatment had little influence on algal fatty acid profiling, hence it did not affect the cetane number that is a key parameter of biodiesel quality. Taken together, our evidence supports a chemically enhanced lipid production of microalgae under low sub-optimal temperatures whose lipid output was comparable to those at optimal temperatures. The novel application of glycine betaine could strengthen the algal biodiesel industry by extending the geographical range for cultivation into areas with cooler climates.

Betaine increases the butyrylcholinesterase activity in rat plasma.

The physiological function of butyrylcholinesterase (EC 3.1.1.8, BChE) is not clearly understood, but a role was suggested in the fat utilization process, resulting in positive correlation between plasma triglyceride (TG) levels and BChE activity. Consequently we tested the hypothesis that regular intake of betaine, a natural compound intervening in the liver TG metabolism could influence the BChE activity. The BChE activity was estimated spectrophotometrically in plasma of rats fed with betaine enriched standard (B) or high-fat diet (HFB). The results confirmed decreased TG plasma levels after betaine treatment independently on the type of diet (0.15+/-0.03 (B) vs. 0.27+/-0.08 (control) mmol/l; p=0.003 and 0.13+/-0.03 (HFB) vs. 0.27+/-0.08 (control) mmol/l; p=0.005). The BChE activity increased significantly with betaine administration, however the change was more distinct in the HFB group (0.84+/-0.34 (HFB) vs. 0.22+/-0.04 (control) O.D./min/mg; p<0.001 and 0.41+/-0.11 (B) vs. 0.22+/-0.04 (control) O.D./min/mg; p=0.001). In conclusion, betaine intake led to elevated BChE activity in plasma and this effect was potentiated by the HF diet. Since betaine is in general used as a supplement in the treatment of liver diseases accompanied by TG overload, its impact on the BChE activity in the role of the liver function marker should be taken into account.

Exogenous glycine betaine treatment enhances chilling tolerance of peach fruit during cold storage

The effect of exogenous glycine betaine (GB) treatment on chilling injury (CI), energy status, endogenous GB, γ-aminobutyric acid (GABA) and proline content in cold-stored peach fruit was investigated. The results showed that GB treatment significantly prevented CI and maintained low flesh firmness and high extractable juice in peach fruit during cold storage. The accumulation of endogenous GB, GABA and proline contents in peaches was enhanced by GB treatment resulting from increasing activities of metabolism enzymes, including betaine aldehyde hydrogenase (BADH), glutamate decarboxylase (GAD), Δ1-pyrroline-5-carboxylate synthetase (P5CS) and ornithine δ-aminotransferase (OAT). Moreover, high level of energy status in GB-treated peach fruit was found. These results indicated that GB treatment enhanced chilling tolerance in peaches due to induction of energy status, endogenous GB, GABA and proline contents, which prevented membrane damage. Thus, exogenous GB treatment could be a useful method to reduce CI in peach fruit during cold storage.

Betaine for Patients with Acute Lymphoblastic Leukemia Intolerant of Maintenance Chemotherapy Due Deficiency of S-Adenosyl Methionine

Background: We noted that rare patients with acute lymphoblastic leukemia (ALL) who are intolerant to oral mercaptopurine (6MP) and methotrexate (MTX) have low plasma S-adenosyl methionine (SAM) levels of less than70 nM. SAM is the methyl donor for the metabolic detoxification of 6MP. Betaine, an oral supplement can serve as a methyl donor for the restoration of SAM. The purpose of this study was to document the effect of oral betaine on tolerance of oral 6MP and MTX during maintenance therapy in select children with ALL who have repeated interruptions of therapy that may compromise cure.Procedure: The data from five patients with multiple interruptions of maintenance chemotherapy and low SAM seen over a five year period were compared before and after betaine treatment. None of the patients were deficient in thiopurine methyltransferase. One patient was homozygous deficient for MTHFR C667T. These patients continued to have interruptions after switching from sulfamethoxazole-trimethoprim to pentamidine. The following were compared before and after betaine treatment: the percentage of days chemotherapy had to be interrupted, plasma SAM levels, mercaptopurine red cell metabolites, dose of 6MP and MTX, and variance in blood cell counts. Betaine was started at a dose of 25 mg/kg/dose twice a day and increased weekly by 50 mg/kg/day as tolerated to keep SAM >70. Doses up to 150 mg/kg/day have been used in patients with hyperhomocysteinemia.Results: Betaine treatment significantly reduced the percentage of days maintenance chemotherapy was withheld (figure right panel). There was less variability in blood cell counts and increased plasma SAM levels after betaine therapy. Betaine altered neither the average dose of 6MP and MTX nor the red cell 6MP metabolites levels. The figure left panel shows the effect of betaine supplementation in the index patient who had no interruptions and stable blood counts on 6MP (75 mg/m2/day) and MTX (14 mg/m2/week).Conclusions: We concluded that betaine is effective in reducing maintenance chemotherapy interruptions in patients with intolerance due to low SAM. An in vitro study showed SAM depletion occurs from 6MP metabolites due to inhibition of de novo protein synthesis, depleting ATP which is necessary for the conversion of methionine to SAM [Stet EH et al. Biochem. J. 1994:304:163-168]. This may lead to alteration in gene methylation possibly resulting in another mechanism of leukemic killing by 6MP. While this finding is concerning, non-adherence to oral 6MP is a known risk for relapse. Betaine allows the rare patient with maintenance intolerance due to SAM deficiency to continue on stable doses of 6MP and MTX without interruption.Left Panel: Hemoglobin, ANC and platelet count for patient #1 before and after starting betaine on day 570.Right Panel: Percent days oral 6MP and MTX withheld during maintenance before (left bar, blue) and after (right bar, red) betaine supplementation (p = 0.02). [Display omitted] [Display omitted] DisclosuresOff Label Use: Betaine is FDA approved for hyperhomocysteinemia. It is also available as a supplement. We are using it to increase deficient S-adenosyl methionine (SAM) levels..

Betaine reverses the memory impairments in a chronic cerebral hypoperfusion rat model

Vascular dementia (VaD) is the second reason for the cognitive decline in aged people, but the effective therapy is still missing. The chronic cerebral hypoperfusion (CCH) had been widely found in VaD patients and is thought to be the key reason for cognitive impairment. Betaine is a natural product that had been implicated in many biological processes and had been used for the therapy of some neurodegenerative disease, such as Alzheimer’s disease. In this study, we reported that betaine treatment could rescue the memory deficits induced by two-vessel occlusion (2-VO), a widely used CCH rat model. Betaine also restored the expression of PSD93, PSD95 and MAP2 to preserve the synaptic functions. Furthermore, betaine could reduce the oxidative stress by suppressing the MDA and ROS and enhancing the SOD and GSH. Overall, betaine treatment is able to rescue the memory deficits in CCH rats, which provide an experimental basis for the therapy of VaD.

Postharvest trans-resveratrol and glycine betaine treatments affect quality, antioxidant capacity, antioxidant compounds and enzymes activities of ‘El-Bayadi’ table grapes after storage and shelf life

The effect of exogenous trans-resveratrol (1.6×10−5M, 1.6×10−4M and 1.6×10−3M) and glycine betaine (GB) (10, 15 and 20mM/L) treatments on quality, antioxidant capacity, antioxidant compounds and enzymes activities of ‘El-Bayadi’ table grapes after 30 days of cold storage plus 2 days of shelf life were evaluated. trans-resveratrol especially at medium and high rates decreased decay after storage compared to control. GB, only at the high rate, decreased decay. Weight loss was higher at the low rate of trans-resveratrol than control. However, GB had no effect on weight loss. TSS, acidity and pH slightly changed during storage and were not affected by trans-resveratrol and GB. Firmness decreased after storage compared to initial, but was higher at trans-resveratrol and GB treatments than control. Both trans-resveratrol and GB retained higher total phenols concentration after storage than control. The high rate of trans-resveratrol and GB retained higher total flavonoids concentration than control. Vitamin C concentration was not affected by the applied treatments. Both trans-resveratrol and GB increased peroxidase (POD) and polyphenoloxidase (PPO) activities compared to control. Polygalacturonase (PG) activity greatly decreased by trans-resveratrol treatments. Xylanase activity increased by the medium rate of trans-resveratrol and the high rate of GB. Antioxidant capacity measured by DPPH assay decreased after storage in control compared to initial, but was higher at trans-resveratrol than other treatments. GB at low rate showed higher antioxidant activity than control. Also, antioxidant capacity measured by ABTS assay decreased after storage in control compared to initial, but was higher at the high rate of trans-resveratrol than other treatments. It is concluded that exogenous trans-resveratrol and GB treatments retained quality of ‘El-Bayadi’ table grapes after cold storage and shelf life and being suggested as natural alternatives to synthetic chemicals.

FAT10 suppression stabilizes oxidized proteins in liver cells: Effects of HCV and ethanol

FAT10 belongs to the ubiquitin-like modifier (ULM) family that targets proteins for degradation and is recognized by 26S proteasome. FAT10 is presented on immune cells and under the inflammatory conditions, is synergistically induced by IFNγ and TNFα in the non-immune (liver parenchymal) cells. It is not clear how viral proteins and alcohol regulate FAT10 expression on liver cells. In this study, we aimed to investigate whether FAT10 expression on liver cells is activated by the innate immunity factor, IFNα and how HCV protein expression in hepatocytes and ethanol-induced oxidative stress affect the level of FAT10 in liver cells. For this study, we used HCV+ transgenic mice that express structural HCV proteins and their HCV− littermates. Mice were fed Lieber De Carli diet (control and ethanol) as specified in the NIH protocol for chronic–acute ethanol feeding. Alcohol exposure enhanced steatosis, induced oxidative stress and decreased proteasome activity in the liversof these mice, with more robust response to ethanol in HCV+ mice. IFNα induced transcriptional activation of FAT10 in liver cells, which was dysregulated by ethanol feeding. Accordingly, IFNα-activated expression of FAT10 in hepatocytes (measured by indirect immunofluorescent of liver tissue) was also suppressed by ethanol exposure in both HCV+ and HCV− mice. This suppression was accompanied with ethanol-mediated induction of lipid peroxidation marker, 4-HNE. All aforementioned effects of ethanol were attenuated by in vivo feeding of mice with the pro-methylating agent, betaine, which exhibits strong anti-oxidant properties. Based on this study, we hypothesize that FAT10 targets oxidatively modified proteins for proteasomal degradation, and that the reduction in FAT10 levels along with decreased proteasome activity may contribute to stabilization of these altered proteins in hepatocytes. In conclusion, IFNα induced FAT10 expression, which is suppressed by ethanol feeding in both HCV+ and HCV− mice. Betaine treatment reverses HCV–ethanol induced dysregulation of protein methylation and oxidative stress, thereby restoring the FAT10 expression on liver cells.

The beneficial effects of betaine on dysfunctional adipose tissue and N6-methyladenosine mRNA methylation requires the AMP-activated protein kinase α1 subunit

The current study was conducted to determine whether betaine could improve fatty acid oxidation, mitochondrial function and N6-methyladenosine (m6A) mRNA methylation in adipose tissue in high-fat-induced mice and how AMP-activated protein kinase α1 subunit (AMPKα1) was involved. AMPKα1 knockout mice and wild-type mice were fed either a low-fat diet, high-fat diet or high-fat diet supplemented with betaine in the drinking water for 8weeks. Our results showed that mitochondrial genes (PGC1α) and β-oxidation-related genes (CPT1a) at protein level were increased in wild-type mice supplemented with betaine when compared with those in mice with high-fat diet. Betaine also decreased FTO expression and improved m6A methylation in adipose tissue of wild-type mice with high-fat diet. However, betaine failed to exert the abovementioned effects in AMPKα1 knockout mice. In adipocytes isolated from mice with high-fat diet, betaine treatment increased lipolysis and lipid oxidation. Moreover, betaine decreased FTO expression and increased m6A methylation. However, while AMPKα1 was knockdown, no remarkable changes in adipocytes were observed under betaine treatment. Our results indicated that betaine supplementation rectified mRNA hypomethylation and high FTO expression induced by high-fat diet, which may contribute to its beneficial effects on impaired adipose tissue function. Our results suggested that the AMPKα1 subunit is required for the beneficial effects of betaine on dysfunctional adipose tissue and m6A methylation. These results may provide the foundation for a mechanism that links m6A methylation status in RNA, AMPKα1 phosphorylation and dysfunctional adipose tissue induced by high-fat diet.

Betaine prevents homocysteine-induced memory impairment via matrix metalloproteinase-9 in the frontal cortex

Betaine plays important roles that include acting as a methyl donor and converting homocysteine (Hcy) to methionine. Elevated plasma Hcy levels are known as hyperhomocysteinemia (HHcy) and contribute to impairments of learning and memory. Although it is commonly known that betaine plays an important role in Hcy metabolism, the effects of betaine on Hcy-induced memory impairment have not been investigated. Previously, we demonstrated the beneficial effects of betaine on acute stress and lipopolysaccharide-induced memory impairment. In the present study, we investigated whether betaine ameliorates Hcy-induced memory impairment and the underlying mechanisms of this putative effect. Mice were treated with Hcy (0.162mg/kg, s.c.) twice a day for nine days, and betaine (25mg/kg, s.c.) was administered 30min before the Hcy injections. The memory functions were evaluated using a spontaneous alternation performance test (Y-maze) at seven days and a step-down type passive avoidance test (SD) at nine and ten days after Hcy injection. We found that betaine suppressed the memory impairment induced by repeated Hcy injections. However, the blood concentrations of Hcy were significantly increased in the Hcy-treated mice immediately after the passive avoidance test, and betaine did not prevent this increase. Furthermore, Hcy induces redox stress in part by activating matrix metalloproteinase-9 (MMP-9), which leads to BBB dysfunction. Therefore, we tested whether betaine affected MMP-9 activity. Interestingly, treatment with betaine significantly inhibited Hcy-induced MMP-9 activity in the frontal cortex but not in the hippocampus after acute Hcy injection. These results suggest that the changes in MMP-9 activity after betaine treatment might have been partially responsible for the amelioration of the memory deficits and that MMP-9 might be a candidate therapeutic target for HHcy.

Betaine inhibits vascularization via suppression of Akt in the retinas of streptozotocin-induced hyperglycemic rats.

Diabetic retinopathy is a severe microvascular complication amongst patients with diabetes, and is the primary cause of visual loss through neovascularization. Betaine is one of the components of Fructus Lycii. In the present study, the effects of betaine on the expression levels of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor (HIF)-1α in association with the Akt pathway were investigated in the retinas of streptozotocin (STZ)-induced diabetic rats using western blot and immunohistochemical analyses. The results of the present study revealed that the expression levels of VEGF, HIF-1α, and Akt were increased in the retinas of the STZ-induced diabetic rats. Betaine treatment attenuated this increase in VEGF and HIF-1α expression via suppression of diabetes-induced Akt activation in the retinas of the diabetic rats. The results suggested that betaine may potentially be used to delay the onset of complications associated with diabetic retinopathy via inhibition of retinal neovascularization in patients with diabetes.

Newborn screening for homocystinurias and methylation disorders: systematic review and proposed guidelines.

Newborn screening (NBS) is justified if early intervention is effective in a disorder generally not detected early in life on a clinical basis, and if sensitive and specific biochemical markers exist. Experience with NBS for homocystinurias and methylation disorders is limited. However, there is robust evidence for the success of early treatment with diet, betaine and/or pyridoxine for CBS deficiency and good evidence for the success of early betaine treatment in severe MTHFR deficiency. These conditions can be screened in dried blood spots by determining methionine (Met), methionine-to-phenylanine (Met/Phe) ratio, and total homocysteine (tHcy) as a second tier marker. Therefore, we recommend NBS for cystathionine beta-synthase and severe MTHFR deficiency. Weaker evidence is available for the disorders of intracellular cobalamin metabolism. Early treatment is clearly of advantage for patients with the late-onset cblC defect. In the early-onset type, survival and non-neurological symptoms improve but the effect on neurocognitive development is uncertain. The cblC defect can be screened by measuring propionylcarnitine, propionylcarnitine-to-acetylcarnitine ratio combined with the second tier markers methylmalonic acid and tHcy. For the cblE and cblG defects, evidence for the benefit of early treatment is weaker; and data on performance of Met, Met/Phe and tHcy even more limited. Individuals homozygous or compound heterozygous for MAT1A mutations may benefit from detection by NBS using Met, which on the other hand also detects asymptomatic heterozygotes. Clinical and laboratory data is insufficient to develop any recommendation on NBS for the cblD, cblF, cblJ defects, glycineN-methyltransferase-, S-adenosylhomocysteinehydrolase- and adenosine kinase deficiency.

Beneficial antioxidant properties of betaine against oxidative stress mediated by levodopa/benserazide in the brain of rats.

The present study was designed to evaluate antioxidant effects of betaine in the brain following administration of levodopa and benserazide, which are routinely used in the treatment of Parkinson's disease. Sprague-Dawley male rats were divided into levodopa (LD), Betaine (Bet.), levodopa plus betaine (LD/Bet.), levodopa plus benserazide (LD/Ben.), levodopa plus betaine-benserazide (LD/Bet.-Ben.) and control groups. The experimental groups received LD 300mg/kg, Bet. 1.5% w/w of the total diet, Ben. 75mg/kg and distilled water to controls for 10 consecutive days, orally. The concentration of plasma total homocysteine significantly increased in LD/Ben.-treated rats when compared to the other groups. Brain glutathione peroxidase (GPx) activity and glutathione content both elevated with betaine treatment in LD/Bet. and LD/Bet.-Ben groups. Superoxide dismutase activity was also higher in controls and betaine-treated rats in comparison with LD and LD/Ben. groups. Likewise, catalase activity significantly increased in control and betaine groups when compared to LD- and LD/Ben.-treated rats. In contrast, brain lipid peroxidation significantly increased in response to LD and LD/Ben. Regarding metabolism of LD in peripheral tissues, serumic dopamine concentration significantly increased in LD-treated rats in comparison with LD/Ben. group. The present results show beneficial antioxidant and methyl donor properties of betaine versus oxidative stress and hyperhomocysteinemia induced by levodopa and benserazide in an animal model.

Effects of in Ovo Administration of Betaine and Choline on Hatchability Results, Growth and Carcass Characteristics and Immune Response of Broiler Chickens

The effect of in ovo administration of different levels of betaine and choline on egg hatchability, immune response, growth and carcass traits of broiler chickens was studied. Four thousand hatching eggs from Ross 308 broiler breeder layers, weighed individually, were incubated for 21 days in a commercial hatchery. At 12th day of incubation, 3456 fertilized eggs were randomly divided into 8 experimental groups of 3 replicates each (144 eggs per replicate): negative control (NC) – not injected; positive control (PC) – injected with 0.5 mL deionized water; Bet 0.25 – injected with 0.5 mL deionized water+0.25 mg soluble betaine; Bet 0.375 – injected with 0.5 mL deionized water+0.375 mg soluble betaine; Bet 0.50 – injected with 0.5 mL deionized water+0.50 mg soluble betaine; Chol 0.25 – injected with 0.5 mL deionized water+0.25 mg soluble choline; Chol 0.375 – injected with 0.5 mL deionized water+0.375 mg soluble choline; Chol 0.50 – injected with 0.5 mL deionized water+0.50 mg soluble choline. Among the hatched chickens, 360 males were randomly chosen (45 for each group) and were grown up to 42nd day of age. The embryo mortality, pecked eggs, infected eggs and hatchability percentages were similar among the experimental groups. The betaine and choline treatments improved hatching weight and final weight of chickens, while reduced feed conversion ratio and abdominal fat percentage. No effect on carcass yield, and breast muscle, leg and wings percentages, as well as on immunoglobulin M (IgM), G (IgG), and total antibody (IgT) titers was observed. The treatments had little effect on internal organs.

Protective effect of betaine against burn-induced pulmonary injury in rats.

This study was designed to determine possible protective effect of betaine treatment against oxidative injury in pulmonary tissue induced with thermal trauma. Under ether anesthesia, shaved dorsum of Wistar albino rats was exposed to a 90°C water bath for 10 seconds to induce burn injury. Betaine was administered orally (250 mg/kg) for a period of 21 days before burn injury, and single dose of betaine was administered after thermal injury. Control group rats were exposed to 25°C water bath for 10 seconds. Upon conclusion of experiment, rats were decapitated and blood was collected for analysis of pro-inflammatory cytokines and lactate dehydrogenase (LDH) activity. Lung tissue samples were taken to determine malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO), and Na+/K+-ATPase activity, in addition to histological analysis. Burn injury caused significant increase in both cytokine levels and LDH activity. In lung samples, raised MDA levels, MPO activity, and reduced GSH levels and Na+/K+-ATPase activity were found due to burn injury. Treatment of rats with betaine significantly restored GSH level and Na+/K+-ATPase activity, and decreased MDA level and MPO activity. According to the findings of the present study, betaine significantly diminishes burn-induced damage in tissue.

Betaine is a positive regulator of mitochondrial respiration

Betaine protects cells from environmental stress and serves as a methyl donor in several biochemical pathways. It reduces cardiovascular disease risk and protects liver cells from alcoholic liver damage and nonalcoholic steatohepatitis. Its pretreatment can rescue cells exposed to toxins such as rotenone, chloroform, and LiCl. Furthermore, it has been suggested that betaine can suppress cancer cell growth in vivo and in vitro. Mitochondrial electron transport chain (ETC) complexes generate the mitochondrial membrane potential, which is essential to produce cellular energy, ATP. Reduced mitochondrial respiration and energy status have been found in many human pathological conditions including aging, cancer, and neurodegenerative disease. In this study we investigated whether betaine directly targets mitochondria. We show that betaine treatment leads to an upregulation of mitochondrial respiration and cytochrome c oxidase activity in H2.35 cells, the proposed rate limiting enzyme of ETC in vivo. Following treatment, the mitochondrial membrane potential was increased and cellular energy levels were elevated. We propose that the anti-proliferative effects of betaine on cancer cells might be due to enhanced mitochondrial function contributing to a reversal of the Warburg effect.