Methylenetetrahydrofolate Research Articles

Development and Validation of RP-HPLC Method for the Estimation of L-Methyl Folate and Tofisopam in combined Formulation

Development and Validation of RP-HPLC Method for the Estimation of L-Methyl Folate and Tofisopam in combined Formulation

Comparative Study between the Use of Regular Folic Acid Supplement versus the Use of L-Methyl Folate in Patients with Methyl Tetrahydrofolate Reductase (MTHFR) Gene Mutation with Recurrent Pregnancy Loss

Background: Methylfolate is the active metabolite of folate that is important for DNA repair, synthetized under the effect of MTHFR (methyl-tetrahydro-folatereductase) enzyme. Patients with MTHFR gene mutation have low levels of biologically active methyfolate. Those patients have high homocysteine levels causing vasculopathy and inadequate feto-maternal circulation. Aim of the Work: To predict the potential benefit of use of methylfolate instead of use of the regular folic acid in patients with MTHFR gene mutation with history of RPL (recurrent pregnancy loss). Subjects and Methods: Study was performed on 100 women. All women had experienced at least two consecutive miscarriages first trimester abortion. All patients were positive of having MTHFR gene mutation. Patients were divided into two groups in terms of 1st trimester drug intake. The 1st group recieved a regular folic acid supplement in a dose of 5 mg per day starting from the day of positive pregnancy test till the end of the first trimester. The 2nd group recieved L-methylfolate supplement in a dose of 1000 mcg per day starting from the day of positive pregnancy test till the end of the first trimester. Then both groups were compared in terms of abortion rates, pregnancy continuation rates and the development of other major obstetric complications. Results: Patients in 1st group had no associated pregnancy related complications in 56%, PE in 14%, PROM in 18% and PTL in 12% of cases. On the other hand, patients in 2nd group had no associated pregnancy related complications in 78%, PE in 6%, PROM in 8% and PTL in 8% of cases 54% of patients on folate group ended in abortion, while only 16% of patients on methylfolate group had abortion. 24% of patients on folate group had PTL, compared to 8% of patient who had had PTL in methylfolate group. 22% of patients in the 1st group continued pregnancy to full term, while 60% of 2nd group continued pregnancy to full term. Conclusion and Recommendations: The use of methyl-folate supplement during the first trimester of in patients with history of RPL and positive MTHFR gene mutation should be a routine practice instead of the regular folate supplement as it improves pregnancy continuation rates and decreases occurrence of associated pregnancy co-morbidities as preterm labor and preeclampsia.

Pregnancy outcomes in women with epilepsy and MTHFR mutations supplemented with methylated folate and methylcobalamin (methylated B12).

Antiseizure medications (ASM) may contribute to adverse fetal outcomes in pregnant women with epilepsy (WWE). Folate processing (Methylenetetrahydrofolate reductase, MTHFR) gene abnormalities are common in women with epilepsy and depression. L-methylfolate supplements may bypass MTHFR deficiencies, yet their use in WWE during gestation or on fetal development is not well studied. We examine pregnancy histories of three WWE who supplemented with either folate or L-methylfolate and methylcobalamin (methylated B12) during pregnancies. Their pregnancy outcomes improved with L-methylfolate and methylcobalamin supplementation. L-methylfolate and methylcobalamin supplementation merits further study in WWE who have MTHFR mutations, fertility, recurrent miscarriage and or depression histories.

First case report of cerebral folate deficiency caused by a novel mutation of FOLR1 gene in a Chinese patient

BackgroundCerebral folate deficiency (CFD) is a neurological disease, hallmarked by remarkable low concentrations of 5-methyltetrahydrofolic acid (5-MTHF) in cerebrospinal fluid (CSF). The primary causes of CFD include the presence of folate receptor (FR) autoantibodies, defects of FR encoding gene FOLR1, mitochondrial diseases and congenital abnormalities in folate metabolism.Case presentationHere we first present a Chinese male CFD patient whose seizure onset at 2 years old with convulsive status epilepticus. Magnetic Resonance Imaging (MRI) revealed the development of encephalomalacia, laminar necrosis in multiple lobes of the brain and cerebellar atrophy. Whole Exome Sequencing (WES) uncovered a homozygous missense variant of c.524G > T (p.C175F) in FOLR1 gene. Further laboratory tests demonstrated the extremely low level of 5-MTHF in the CSF from this patient, which was attributed to cerebral folate transport deficiency. Following the intravenous and oral treatment of calcium folinate, the concentrations of 5-MTHF in CSF were recovered to the normal range and seizure symptoms were relieved as well.ConclusionsOne novel variation of FOLR1 was firstly identified from a Chinese male patient with tonic-clonic seizures, developmental delay, and ataxia. The WES and laboratory results elucidated the etiology of the symptoms. Clinical outcomes were improved by early diagnosis and proper treatment.

Analysis OF C677T polymorphism in methylene tetrahydrofolate reductase (MTHFR) gene as a risk factor for congenital talipes equino varus (CTEV)

BackgroundClubfoot is a common congenital foot deformity. Low folate status in mothers has been associated with CTEV. Folate metabolism might be affected by Methylene Tetrahydrofolate Reductase (MTHFR) gene polymorphism. The present study was aimed to investigate MTHFR C677T polymorphism and its association with CTEV. MethodsThis is a Case-mother-Dyad study with 30 pairs of cases and controls. Single Nucleotide Polymorphism (SNP) analysis of the MTHFR gene was done in this hospital-based study by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). ResultsIn this study, we observed less relative risk of CTEV in presence of C allele as compared to T allele in children, with Relative Risk- 0.6281 and likelihood ratio of 0.5714. While analysing the correlation of genotype variation in cases (CC = 8(26.66%) and CT = 22(73.33%)) with there biological mother (CC = 13(43.33%) and CT = 17(56.66%)), no significant correlation (p = 0.3110) was found between cases and their biological mother genotype. ConclusionAmong the enrolled cases, there was a significant association of increased CTEV risk with 677T variant allele of MTHFR gene. Also, maternal MTHFR genotype was not found to influence CTEV risk of offspring.

A prospective birth cohort study on cord blood folate subtypes and risk of autism spectrum disorder

We previously reported that extremely high concentrations of maternal plasma folate were associated with increased risk of autism spectrum disorder (ASD) in children. This study explored whether specific types of folate in cord blood have differential association with ASD. In the Boston Birth Cohort (BBC), we assessed the association between cord blood unmetabolized folic acid (UMFA), 5-methyl tetrahydrofolate (THF), and total folate and a child's ASD risk. In a subset, we explored whether the association between UMFA and ASD risk can be affected by the dihydrofolate reductase (DHFR) genotype and cord plasma creatinine. We also examined prenatal correlates of cord UMFA concentrations. This report included 567 BBC children (92 ASD, 475 neurotypical), who were recruited at birth and prospectively followed at the Boston Medical Center. ASD was defined from International Classification of Diseases (ICD)-9 and ICD-10 codes documented in electronic medical records. Children with cord UMFA in the highest, versus lowest quartile, had a greater ASD risk (adjusted OR, aORquartile4: 2.26; 95% CI: 1.08, 4.75). When stratified by race/ethnicity, the association was limited to 311 (45 ASD) Black children (aORquartile4: 9.85; 95% CI: 2.53, 38.31); a test of interaction between race/ethnicity and cord UMFA concentrations was significant (P=0.007). The UMFA-ASD association in Black children slightly attenuated after adjusting for cord plasma creatinine (P=0.05). There was no significant association between cord 5-methyl THF, total folate, DHFR genotype, and ASD risk. Cord total folate and maternal supplement intake during second trimester were associated with higher cord UMFA. Higher concentrations of cord UMFA, but not 5-methyl THF or total folate, were associated with a greater risk of ASD in Black children. This study in a preterm-birth-enriched cohort raises more questions than it could answer and underscores the need for additional investigations on the sources and role of cord UMFA in children's neurodevelopmental outcomes and underlying mechanisms.

Epigenetic Modification in Methylene Tetrahydrofolate Reductase (MTHFR) Gene of Women with Pre-eclampsia

Genetic and epigenetic factors play significant roles in the aetio-pathogenesis of pre-eclampsia (PE). The effects may vary across racial and geographical boundaries. The role of epigenetic modification in pre-eclampsia was studied among African populations in Lagos, Nigeria.This study aimed to determine the pattern of Methylene tetrahydrofolate reductase gene (MTHFR) CpG island methylation in pre-eclampsia, and evaluate associated covariates.This study was an observational, cross-sectional, study conducted at the Lagos University Teaching Hospital and the Lagos State Island Maternity Hospital. A total of 400 pregnant women consisting of 200 pregnant women diagnosed with pre-eclampsia (study group) and 200 pregnant normotensive and apparently healthy women (control group) were recruited for the study. Demographic and clinical histories were obtained through questionnaires. The DNA Methylation status of the CpG Island in promoter region of the MTHFR gene was assessed using bisulphite conversion and methylation specific PCR method. The biochemical parameters measured in the study were: red cell folate, vitamin B12, plasma homocysteine (Hcy) and methylene tetrahydrofolate reductase enzyme level.Homozygous MTHFR CpG island hypomethylation pattern was significantly associated with pre-eclampsia (χ2 = 22.96; p = 0.000), Mean values of plasma homocysteine in PE women with homozygous hypomethylation (26.1 ± 9.1 umol/L) were significantly higher than (20.1 ± 4.2 umol/L) observed in PE subjects with homozygous hypermethylation (p = 0.008). Homozygous CpG island hypomethylated pattern of the MTHFR promoter region, was associated with the lowest median MTHFR enzyme level (72.8 ± 39.8 pmol/L) compared with heterozygous methylated pattern (91.3 ± 60.9 pmol/L; p = 0.047) and homozygous methylated pattern (82.3 ± 31.0 pmol/L; 0.047). Red cell folate and Vitamin B12 levels were not significantly associated with CpG island methylation status.Epigenetic modification plays significant role in the pathogenesis of pre-eclampsia.

Predictive biomarkers of IgA vasculitis with nephritis by metabolomic analysis

BackgroundIgA vasculitis (IgAV) is the most common vasculitis of childhood. Renal involvement defines late morbidity of the disease. A better understanding of the pathophysiology of the progression to kidney disease and predictive biomarkers are required for better management of IgAV and its nephritis (IgAVN). ObjectivesAn untargeted metabolomics approach was performed to reveal the underlying molecular mechanism of disease pathogenesis and to define potential biomarkers from plasma samples from IgAV and IgAVN patients. MethodsForty-five active IgAV patients (H) and six healthy controls (C) were enrolled in the study. Plasma samples were collected on the same day of diagnosis and before any immunosuppressive treatment was initiated. At the time of diagnosis and sample collection, none of the patients had renal involvement. We used Quadrupole Time of Flight Mass Spectrometry (Q-TOF LC/MS) to investigate the alterations in plasma metabolomic profiles. Three separate pools were created: healthy controls (group C), active IgAV patients who did not develop renal involvement (group H), and patients who developed IgAVN at follow up (group N). Peak picking, grouping, and comparison parts were performed via XCMS (https://xcmsonline.scripps.edu/) software. ResultsAt follow-up, IgAVN developed in 6 out of 45 IgAV patients. The median time of renal involvement development is 23 days (range 5–45 days). Of these, 3 had nephritic proteinuria, one had nephrotic proteinuria, and 2 had microscopic hematuria. There were no significant differences in gender, age, clinical manifestations, and laboratory findings between the six patients who developed renal involvement and those who did not. In multivariate analysis, there was no significant association between any of the defined demographic and clinical characteristics (male sex, gastrointestinal system involvement, joint involvement, CRP, WBC, PLT) and the occurrence of renal involvement.Totally 2618 peaks were detected for group H, N, and C. Among them, 355 peaks were found to be statistically significant and reliable (p<0.05), and 155 of these peaks were found to be changed (fold change >1.5) between the groups C and H, and 66 peaks were found to be changed (fold change >1.5) between the groups H and N. The number of the peaks on the intersection of the peaks found to be different between the groups (C and H) and (H and N) was 39. Based on putative identification results, 15 putatively identified metabolites matched with 11 peaks were presented as biomarker candidates after careful evaluation with a clinical perspective. ConclusionWe suggest that DHAP (18:0), prostaglandin D2/I2, porphobilinogen, 5-methyltetrahydrofolic acid, and N-Acetyl-4-O-acetylneuraminic acid/N-Acetyl-7-O-acetylneuraminic acid may serve as biomarkers for predicting kidney disease. Future studies with larger groups of IgAV patients are needed to validate the identified metabolic profile.

Development and Validation of Bioanalytical Method for Simultaneous Estimation of Leucovorin and its Primary Metabolite 5-Methyl Tetrahydrofolic Acid from Human Plasma by Liquid Chromatography Tandem Mass Spectrometry with Solid Phase Extraction

A simple, highly sensitive, precise and accurate liquid chromatographic mass spectroscopic (LC-MS/MS) method has been developed and validated for simultaneous quantification of Leucovorin and its metabolite 5-Methyl tetrahydrofolic acid in human plasma using Leucovorin-d4 and 5-Methyl-d3 tetrahydrofolic acid as internal standard (IS), respectively. The analytes were extracted from human plasma by solid phase extraction (SPE) technique using ion exchange cartridges. Chromatographic separation was achieved with reversed phase column chromatography. API 5500 LC-MS/MS in multiple reactions monitoring (MRM) was used for detection. Obtained results revealed that the method had a short sample preparation time as well as a chromatographic run time of just 3.0 min. In summary our results suggest that the validated method fulfilled all preset criteria for sensitivity, specificity, linearity (5.0-700.0 ng/mL), inter-and intra-accuracy and precision for both molecules.

Improved conjunctival microcirculation in diabetic retinopathy patients with MTHFR polymorphisms after Ocufolin™ Administration

PurposeTo investigate conjunctival microvascular responses in patients with mild diabetic retinopathy (MDR) and methylenetetrahydrofolate reductase (MTHFR) polymorphisms (D + PM) after administration of Ocufolin™, a medical food containing 900 μg l-methylfolate (levomefolate calcium or [6S]-5-methyltetrahydrofolic acid, calcium salt), methylcobalamin, and other ingredients. MethodsEight D + PM patients received Ocufolin™ for six months (6 M). Bulbar conjunctival microvasculature and microcirculation metrics, including vessel diameter (D), axial blood flow velocity (Va), cross-sectional blood flow velocity (Vs), flow rate (Q), and vessel density (VD, Dbox), were measured at baseline, 4 M, and 6 M. ResultsThe mean age was 54 ± 7 years. No significant demographic differences were found. Conjunctival microcirculation, measured as Va, Vs, and Q was significantly increased at 4 M and 6 M, compared to baseline. Va was 0.44 ± 0.10 mm/s, 0.58 ± 0.13 mm/s, 0.59 ± 0.13 mm/s in baseline, 4 M, and 6 M, respectively (P < 0.01). Similarly, Vs was 0.31 ± 0.07 mm/s, 0.40 ± 0.09 mm/s, 0.41 ± 0.09 mm/s in baseline, 4 M, and 6 M, respectively (P < 0.05). Q was 107.8 ± 49.4 pl/s, 178.0 ± 125.8 pl/s, 163.3 ± 85.8 mm/s in baseline, 4 M, and 6 M, respectively (P < 0.05). The VD at 6 M was significantly higher than that at baseline (P = 0.017). Changes of D were positively correlated with changes of Va, Q, and VD. Effects of MTHFR and haptoglobin polymorphisms on the improvements of conjunctival microcirculation and microvasculature were found. ConclusionsOcufolin™ supplementation improves conjunctival microcirculation in patients with diabetic retinopathy and common folate polymorphisms.

Medical Decision Support to Reduce Unwarranted Methylene Tetrahydrofolate Reductase (MTHFR) Genetic Testing.

Most major national medical associations have advised against routine MTHFR testing since at least 2013. However, many providers continue to order this unwarranted genetic test. This study assessed the efficacy of an electronic best practice alert to aid ordering providers. We tracked the rate of MTHFR tests ordered per 1 million patients in the twelve months before and after the implementation of an alert that suggested an alternative test. Associated factors including the ordering department, diagnosis, patient sex, and patient age were also analyzed. Chi square analysis was used to compare the difference between pre- and post-alert test ordering rates. A total of 997 MTHFR analysis were ordered in Southern California Kaiser Permanente from January 2017 through December 2018. Overall, the average MTHFR monthly test ordering rates dropped significantly from 12.93 per million patients in 2017 to 7.08 per million patients in 2018 (p = 0.0056). However, testing rates in children were unchanged and, in some associated diagnoses, such as psychiatric illnesses and neurodevelopmental conditions, the testing rates increased. Recommending an alternate test in lieu of the unwarranted one significantly reduced the overall rate of MTHFR testing. The alert was most effective for specialties and diagnoses where MTHFR was historically medically indicated. This suggests such alerts are an effective intervention that health care systems can implement to serve as an educational update and to reduce unwarranted genetic testing.

Methylene tetrahydrofolate reductase C677T polymorphism in Korean livedoid vasculopathy patients

Methylene tetrahydrofolate reductase C677T polymorphism in Korean livedoid vasculopathy patients

Methylene Tetrahydrofolate Reductase Deficiency.

5,10-Methylene-tetrahydrofolate reductase (MTHFR) deficiency is a rare, autosomal recessive, metabolic disorder of folate metabolism, which affects homocysteine remethylation. Elevated homocysteine with normal or low methionine level is the key to diagnosis. Early recognition and treatment with betaine, has been shown to improve the survival and neurological outcomes. The authors report five Indian patients from three unrelated families, with MTHFR deficiency to emphasize the importance of early recognition and initiation of specific treatment.

Methylene Tetrahydrofolate Reductase Gene Polymorphism C677T &amp; Risk of Type two Diabetes Mellitus in Beni-Suef Governorate

Background: Methylene Tetrahydrofolate Reductase (MTHFR) is known to be a regulatory enzyme of homocysteine metabolism. In many ethnic groups point mutations in MTHFR gene are implicated in the pathogenesis of diabetic nephropathy (DN) and other complications of type II diabetes mellitus (T2DM). Aim of the work: The aim of this study was to assess if MTHFR C677T gene polymorphism was one of the risk factors for the development of diabetic nephropathy as well as other complications in type 2 diabetes mellitus patients. Subjects and methods: The MTHFR C6777T polymorphisms were detected in 50 persons by PCR-RFLP. Subjects were divided into 2 groups; 30 patients with type II DM and 20 persons who were non-diabetic healthy controls. Results: The presence of MTHFR 677T allele did not increase the risk of complications in T2DM patients. The presence of mutant genotypes CT and TT did not increase the risk of nephropathy or other complications in the subjects having a C677T mutation. Conclusion: Methylene Tetrahydrofolate Reductase (MTHFR) C677T gene mutation was neither associated with an increased risk of diabetic nephropathy nor other complications of type II diabetes mellitus.

Development of Simple Method for the Determination of 5-Methyl Tetrahydrofolate in Dried Blood Spot by High-Performance Liquid Chromatography

Folate (Reduced) or folic acid (oxidized), is a water-soluble vitamin necessary for thymidylate synthase, methionine synthase, serine hydroxymethyltransferase, and folate deficiency associated with megaloblastic anemia, neural tube defect, and coronary heart disease. The available folate data is minimal due to a dearth of suitable, accurate, field-friendly blood collection and estimation methods. Conventional assays like microbiological and competitive protein-binding radioassay methods are used to estimate the total folate in serum and whole blood. In erythrocytes, folate exists mainly as 5-methyltetrahydrofolate polyglutamate and indicates the long-term storage of folate status. Therefore, the study aimed to develop a simple method for estimating 5-methyl tetrahydrofolate (5-MTHF) by high-performance liquid chromatography (HPLC) in dried blood spots (DBS) as an indicator of folate status. The present study results indicate standard 5-methyltetrahydrofolate with different concentrations with linear regression was showed R2 -0.98, and the minimum detection limit of a standard was 20 pg. The inter-and intra-assay variations were found to be less than 10%. A good correlation (R2 -0.964) between the DBS and blood and the recovery of added standard in the DBS sample was more than 80%. A minimum of three DBS punches equivalent to 20mL of blood was required for the analysis, and the competitive protein-binding radioassay showed slightly high folate compared to the HPLC method. The stability of 5-MTHF in DBS was also tested periodically. DBS technique and HPLC method are simple, sensitive, not expansive, and safe compared to the microbiological and competitive binding assay for assessing sub-clinical folate status in a field survey on a large population.

Serine Catabolism Feeds NADH when Respiration Is Impaired

NADH provides electrons for aerobic ATP production. In cells deprived of oxygen or with impaired electron transport chain activity, NADH accumulation can be toxic. To minimize such toxicity, elevated NADH inhibits the classical NADH-producing pathways: glucose, glutamine, and fat oxidation. Here, through deuterium-tracing studies in cultured cells and mice, we show that folate-dependent serine catabolism also produces substantial NADH. Strikingly, when respiration is impaired, serine catabolism through methylene tetrahydrofolate dehydrogenase (MTHFD2) becomes a major NADH source. In cells whose respiration is slowed by hypoxia, metformin, or genetic lesions, mitochondrial serine catabolism inhibition partially normalizes NADH levels and facilitates cell growth. In mice with engineered mitochondrial complex I deficiency (NDUSF4-/-), serine's contribution to NADH is elevated, and progression of spasticity is modestly slowed by pharmacological blockade of serine degradation. Thus, when respiration is impaired, serine catabolism contributes to toxic NADH accumulation.

Intérêt de la mise en évidence des isoformes de la MTHFR (Méthylènetétrahydrofolate réductase) en infertilité du couple

ObjectiveMTHFR SNPs (Methylene Tetrahydrofolate reductase Single Nucleotide polymorphisms) are biochemical modifications decreasing the capacity to form 5 MTHF 5 methyltetrahydrofolate (5MTHF). Their presence reduces the capacity of the One Carbon cycle, and so the regeneration of Homocysteine (Hcy) and in fine strongly perturbs all the methylation processes. As methylation processes are major regulators in gametogenesis and embryogenesis. We have determined the prevalence of the 2 most important SNPs A1298C and C677T in our population of patients consulting for infertility. MethodsDetermination of the MTHFR SNPs A1298C and C677T, by hybridization using the LAMP Human MTHFR mutation KITs. ResultsOnly 15.8% of our patients (861) do not carry any SNP (WT wid type). Close to 20% of the patients are homozygotes for one mutation or the other. A total of 19.7% are composite heterozygous. A total of 43% of our population is considered “at risk”, based on observations collected for the repeat miscarriages. ConclusionsDetermination of the 2 major MTHFR SNPs is not a “first row” choice, but it must not be neglected and should be carried out in case of repeat ART failures and repeat miscarriages. Some simple therapeutic options can be proposed: they are based on the use of 5MTHF (5MethyleneTetraHydroFolate) the compound downstream the MTHFR.

Methylene tetrahydrofolate reductase and methionine synthase gene polymorphisms as genetic determinants of pre-eclampsia.

Pre-eclampsia (PE) is a leading cause of maternal and neonatal mortality in Africa; and has been associated with the interplay of genetic, metabolic and environmental factors. Polymorphisms of methylene tetrahydrofolate reductase (MTHFR) and methionine synthase (MTR) folate cycle genes, have been controversially associated with pre-eclampsia in studies from different human populations. To determine the distribution of MTHFR C677T and MTR A2756G polymorphisms in a Nigerian population and evaluate possible associations with the occurrence of pre-eclampsia and homocysteine metabolic derangement. This study was a hospital based study carried out in Lagos, South-western Nigeria. Two hundred pregnant women clinically diagnosed with pre-eclampsia (study group) and 200 apparently healthy non-pre-eclamptic pregnant women (control group) were recruited for the study after written informed consent. Pre-eclampsia was diagnosed based on the International Society for the Study of Hypertension in Pregnancy re-classification of 2013. MTHFR C677T and MTR A2756G polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Statistical analyzes were performed using SPSS version 23. Hardy-Weinberg distribution were tested with χ2 test. Logistic regression model was used to evaluate the relationship of variables with pre-eclampsia. A value of p<0.05 was considered statistically significant. MTHFR genotype frequencies of CC, CT and TT were 59.8%; 31.2% and 9.0% in study group and 76.6%; 22.3% and 1.0% in the control group respectively. MTR A2756G genotype frequencies of AA, AG and GG genotypes were 71.9%; 20.1% and 8.0% for the study group and 81.5%; 16.4% and 2.1% for the control group. Occurrence of pre-eclampsia was significantly associated with presence of T allele of MTHFR (OR=1.855; p<0.05) and G allele of MTR genes (OR=1.269; p<0.05), Homozygosity of TG haplotype significantly increased the occurrence of pre-eclampsia among Nigerian women (OR=2.252; p<0.05). Population attributable risk fraction percent for the T and G alleles were 16.4% and 11.5% respectively. Mean plasma Hcy level was not, however, significantly affected by MTHFR/MTR haplotypes (F=1.54; p=0.157). MTHFR C677T and MTR A2756G polymorphisms were associated with pre-eclampsia in a population of pregnant women in Lagos, Nigeria.

Effects of Methylene Tetrahydro Folate Reductase Gene Polymorphisms on Methotrexate Toxicity in Egyptian Pediatric Acute Lymphocytic Leukaemia Patients.

This study was designed to evaluate the effect of Methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms on MTX toxicity in pediatric Egyptian ALL patients. Ninety-Four of Pediatric ALL patients aged 3–13 years (7.6 ± 3.6) on oral maintenance dose of 50 mg/m2 weekly of MTX. MTHFR c.677C>T (rs1801133) and c.1298A>C (rs1801131) genotyping were performed using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). The allele frequencies of c.677C>T were 42.6%, 46.8%, and 10.6% for CC, CT, and TT respectively, while c.1298A>C alleles frequencies were 62.7%, 24.5%, and 12.8% for AA, AC, and CC respectively. None of the investigated polymorphism (C677T or A1298C alleles) was associated with either overall or site specific MTX toxicity regarding anemia (p = 0.99) (p = 0.4), platelets (p = 0.4) (p = 0.4), hepatotoxicity (p = 0.4) (p = 0.7), respectively. The results indicated that between c.677C>T genotypes, CC/CT and TT were associated with hematopoietic toxicities 60.7% and 60% (p = 0.2); platelet toxicities 76.2% and 80% (p = 1) and, hepatotoxicities 40.5% and 60% (p = 0.3), respectively. In the c.1298A>C genotypes, CC/AC and AA presented hematopoietic toxicities 68.6% and 55.9% (p = 0.2), platelet toxicities 82.9% 72.9% (p = 0.3) and, hepatotoxicity 37.1% and 45.8% (p = 0.5), respectively. No significant associations were detected between MTHFR c.677C>T or c.1298A>C polymorphisms and either overall or site specific MTX toxicity.

Mutational landscape screening of methylene tetrahydrofolate reductase to predict homocystinuria associated variants: An integrative computational approach

Methylene tetrahydrofolate reductase (MTHFR) is a flavoprotein, involved in one-carbon pathway and is responsible for folate and homocysteine metabolism. Regulation of MTHFR is pivotal for maintaining the cellular concentrations of methionine and SAM (S-adenosyl methionine) which are essential for the synthesis of nucleotides and amino acids, respectively. Therefore, mutations in MTHFR leads to its dysfunction resulting in conditions like homocystinuria, cardiovascular diseases, and neural tube defects in infants. Among these conditions, homocystinuria has been highly explored, as it manifests ocular disorders, cognitive disorders and skeletal abnormalities. Hence, in this study, we intend to explore the mutational landscape of human MTHFR isoform-1 (h.MTHFR-1) to decipher the most pathogenic variants pertaining to homocystinuria. Thus, a multilevel stringent prioritization of non-synonymous mutations in h.MTHFR-1 by integrative machine learning approaches was implemented to delineate highly deleterious variants based on its pathogenicity, impact on structural stability and functionality. Subsequently, extended molecular dynamics simulations and molecular docking studies were also integrated in order to prioritize the mutations that perturbs structural stability and functionality of h.MTHFR-1. In addition, displacement of Loop (Arg157-Tyr174) and helix α9 (His263-Ser272) involved in open/closed conformation of substrate binding domain were also probed to confirm the functional loss. On juxtaposed analysis, it was inferred that among 126 missense mutations screened, along with known pathogenic mutations (H127 T, A222 V, T227 M, F257 V and G387D) predicted that W500C, P254S and D585 N variants could be potentially driving homocystinuria. Thus, uncovering the prospects for inclusion of these mutations in diagnostic panels based on further experimental validations.