C677T methylenetetrahydrofolate reductase and plasma homocysteine levels among Thai vegans and omnivores.

Hyperhomocysteinemia among vegetarians and vegans is caused mostly by vitamin B12 deficiency. A C-to-T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene results in a thermolabile MTHFR, which may affect homocysteine (Hcy) levels. The importance of this gene mutation among populations depends on the T allele frequency. Blood Hcy, vitamin B12, folate, vitamin B6, and MTHFR C677T mutation status were determined in 109 vegans and 86 omnivores aged 30 - 50 years. The vegans had significantly higher Hcy levels than the omnivores, geometric means (95 % CI) 19.2 (17.0 - 21.7) µmol/L vs. 8.53 (8.12 - 8.95) µmol/L, p < 0.001. A C-to-T mutation in the vegans increased plasma Hcy, albeit insignificantly; geometric means 18.2 µmol/L, 20.4 µmol/L, and 30.0 µmol/L respectively in CC, CT, and TT MTHFR genotypes. There was also a significant decrease in serum folate; geometric means 12.1 ng/mL, 9.33 ng/mL, and 7.20 ng/mL respectively, in the CC, CT, and TT mutants, p = 0.006, and particularly, in the TT mutant compared with the CC wild type, 7.20 ng/mL vs. 12.1 ng/mL, p = 0.023. These findings were not seen in the omnivores. It was concluded that hyperhomocysteinemia is prevalent among Thai vegans due to vitamin B12 deficiency. C-to-T MTHFR mutation contributes only modestly to the hyperhomocysteinemia.

Genetic polymorphisms in Thai neonates with hyperbilirubinemia.

AIM: Polymorphisms of the UGT1A1 gene, SLCO1B1 gene and GST gene have been associated with significant hyperbilirubinemia. We would like to determine whether the variation of UGT1A1 gene, SLCO1B1 gene and GST gene may play a significant role in neonatal hyperbilirubinemia in Thai infants. METHODS: Ninety-one study subjects (hyperbilirubinemic group) and 86 control subjects were studied. RESULTS: The cause of neonatal hyperbilirubinemia could not be identified in 64 infants (70.3%), ABO blood group incompatibility in 14.3% and Glucose-6-phosphate dehydrogenase (G-6-PD) deficiency in 8.8%. In the hyperbilirubinemic group, 23 of 91 (25.3%) infants demonstrated variant of UGT1A1 at nucleotides (nt) 211 as compared to 6 of 86 (7%) in the control group (p = 0.001). There were no significant differences between groups in the variants UGT1A1 at nt 686, SLCO1B1 gene at nt 388, 463 and the GST gene. Male infants with G-6-PD deficiency were associated with hyperbilirubinemia (21.2% vs. 4.8% in the control group) with an odds ratio (OR) of 5.37 (p = 0.02). The relationship between G-6-PD and variant in UGT1A1 gene at nt 211 could not be determined. CONCLUSION: Thai infants with variant in the UGT1A1 at nt 211 or with G-6-PD deficiency are at higher risk for developing neonatal hyperbilirubinemia.

Clinical usefulness of a HPLC method for simultaneous quantitation of plasma homocysteine and cysteine.

OBJECTIVE: Homocysteine (Hcy) is a risk for vascular occlusion. It is metabolized via remethylation to methionine and transsulfuration to cysteine which has also been related to vascular occlusion. Simultaneous determination of Hcy and cysteine has additional clinical usefulness in providing a presumptive clue to the nature of hyperhomocysteinemia. MATERIAL AND METHOD: A manual HPLC method has been worked out for simultaneous determination of plasma Hcy and cysteine. Concentrations of Hcy were validated with the widely used automated Abbott AxSYM assay. Its usefulness was tested in 87 omnivores and 111 vegans. RESULTS: Excellent correlation between the values of Hcy was found between the manual HPLC method and the automated Abbott assay. The vegans had significantly higher levels of Hcy but lower levels of cysteine than the omnivores (mean +/- SD, micromol/L 23.6 +/- 18.0 vs. 8.8 +/- 2.1 p < 0.001, 225 +/- 30 vs. 245 +/- 34 p < 0.001, respectively). In contrast, the vegans had significantly lower levels of serum vitamin B12 and plasma vitamin B6 than the omnivores (median values 186 vs 565 pg/ml, p < 0.001; 37.4 vs. 47.4 nmol/L, p < 0.001 respectively). These findings indicate that the hyperhomocysteinemia in the vegans results from impairment of both remethylation and transsulfuration pathways of Hcy secondary to inadequacy of vitamins B12 and B6 respectively. Thus simultaneous determination of Hcy and cysteine is more useful than determination of only Hcy in that it provides a clue to the nature of hyperhomocysteinemia. CONCLUSION: The manual HPLC method and the Abbott assay gave comparable Hcy values, and thus can be used interchangeably. The HPLC method is economical, useful for hospitals with less demand for determination of Hcy, and capable of simultaneously determining cysteine which has implication in clinical practice.

Folate pathway genetic polymorphisms and susceptibility of central nervous system tumors in Thai children.

BACKGROUND: Folate is an important micronutrient molecule participating in DNA synthesis, methylation and repair mechanisms. Genetic polymorphisms in folate pathway related enzymes including methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C, methionine synthase (MTR) A2756G, thymidylate synthase (TS) 28-bp tandem repeat, and reduced folate carrier (RFC) G80A have been shown to be associated with increased susceptibility for several cancers. The aim of the present study was to evaluate whether single nucleotide polymorphisms in the genes encoding enzymes of the folate pathway predispose to any CNS tumors in Thai children. METHODS: In the present case-control study, we investigated these polymorphisms in genomic DNA from peripheral blood mononuclear cells in 73 Thai children with various types of central nervous system tumors and in 205 age and sex matched controls. RESULTS: Thirty-one out of 73 patients were diagnosed with glial tumors (astrocytoma, oigodendroglioma and ependymoma), 28 with embryonal CNS tumors (medulloblastoma, pinealoblastoma and primitive neuroectodermal tumor), 13 with germ cell tumors and 1 with meningioma. We found that the homozygous CC allele of MTHFR A1298C conferred an increased risk of embryonal CNS tumors (OR: 3.9; 95% CI: 1.3-11.4, p=0.02). CONCLUSION: Our findings thus suggest that folate metabolism may play a role in the pathogenesis of certain specific subtypes of pediatric brain tumor in Thai children, especially embryonal CNS tumors.