Folate Reductase Research Articles

Role of Nrf2/HO-1, PPAR-γ, and cytoglobin signals in the pathogenesis of methotrexate-induced testicular intoxication in rats and the protective effect of diacerein.

Methotrexate (MTX) is an inhibitor of folic acid reductase used in managing a variety of malignancies. Testicular injury by MTX is one of its serious adverse effects. The current investigation aims to assess the protective effects of diacerein (DIA) on testicular injury by MTX and clarify the possible underlying mechanisms. Testicular injury in rats was induced by a single injection of 20mg/kg body weight of MTX. DIA was given in 25mg/kg body weight/day and 50mg/kg body weight/day doses for 10days. Compared to the MTX group, DIA attenuated testicular intoxication as evidenced by improvement of testicular histopathological abnormalities and increased serum testosterone and luteinizing hormone. DIA attenuated testicular oxidative stress changes by lowering testicular MDA and boosting GSH content and SOD activity. Moreover, administration of DIA attenuated MTX-induced testicular inflammation, as proved by decreased TNF-α and IL-6. At the molecular level, DIA induced significant upregulation in Nrf2, HO-1, PPAR-γ, and cytoglobin protein expression. The present results proved that DIA, in a dose-dependent manner, exhibited notable amelioration of testicular toxicity induced by MTX through augmentation of anti-inflammatory and antioxidant effects combined by upregulating Nrf2/HO-1, PPAR-γ, and cytoglobin signaling.

Hybrid PABA-glutamic acid conjugated 1,3,5-triazine derivatives: Design, synthesis, and antimalarial activity screening targeting Plasmodium falciparum dihydro folate reductase enzyme.

Despite the successful reduction in the malaria health burden in recent years, it continues to remain a significant global health problem mainly because of the emerging resistance to first-line treatments. Also because of the disruption in malaria prevention services during the COVID-19 pandemic, there was an increase in malaria cases in 2021 compared to 2020. Hence, the present study outlined the in silico study, synthesis, and antimalarial evaluation of 1,3,5-triazine hybrids conjugated with PABA-glutamic acid. Docking study revealed higher binding energy compared to the originally bound ligand WR99210, predominant hydrogen bond interaction, and involvement of key amino acid residues, like Arg122, Ser120, and Arg59. Fourteen compounds were synthesized using traditional and microwave synthesis. The in vitro antimalarial evaluation against chloroquine-sensitive 3D7 and resistant Dd2 strain of Plasmodium falciparum showed a high to moderate activity range. Compounds C1 and B4 showed high efficacy against both strains and a further study revealed that compound C1 is non-cytotoxic against the HEK293 cell line with no acute oral toxicity. In vivo, study was performed for the most potent antimalarial compound C1 to optimize the research work and found to be effectively suppressing parasitemia of Plasmodium berghei strain in the Swiss albino mice model.

Detection of Methylene Tetrahydrofolate Reductase (MTHFR C677T) Mutation among Acute Lymphoblastic Leukemia in Sudanese Patients.

A genetic polymorphism that causes abnormal folate metabolism may lead to genomic instability and increase susceptibility to malignancies such as Acute Lymphoblastic leukemia (ALL). The purpose of this research is to identify methylene tetrahydrofolate reductase (MTHFR C677T) (NCBI ID: 4524) mutation in ALL patients. The study was a descriptive case-control hospital-based study with one hundred Sudanese participants divided equally into fifty (50) Sudanese ALL diagnosed patients as cases and fifty (50) Sudanese individuals as controls. The MTHFR C677T mutant allele was detected using conventional PCR, with the primer sequence of MTHFR C677T F-TGAAGGAAGGTGTCTGCGGGA R-AGGACGGTGCGGTGAGAGTG. The study was conducted from January to March 2023, and samples were collected from the Radiation and Isotops Center at Khartoum Hospital. The investigation revealed that 12 of the 50 patients in the case group (24%) had the MTHFR C677T mutant allele, and the study also revealed that there is significant correlation with the control group. There is no significant relationship between socio-demographic variables and MTHFR mutation detection in ALL patients. Also, the sociodemographic variables predictors of MTHFR mutation among ALL patients adjusted for smoking habit revealed no significant relationship. According to the findings of this study, the mutant allele of the Methylene Tetra Hydro Folate Reductase C677T was detected and demonstrated varying degrees of significance. It was concluded that the MTHFR C677T gene mutation was associated with acute lymphoblastic leukemia in Sudanese patients.

Factor V Leiden (FVL), Prothrombin G20210A and MTHFR C677T Mutations among Patients of Budd-Chiari Syndrome

Abstract Background Many studies ask whether screening Factor V Leiden (FVL), Prothrombin G20210A and Methyl Tetra Hydro Folate Reductase (MTHFR) C677T Mutations should be routinely performed in patients with Budd–Chiari syndrome (BCS). However, the prevalence of these mutations in such patients is substantially variable. This is the first study in clinical pathology department in Ain Shams University to evaluate the importance of this mutational screening in patients with Budd-Chiari syndrome to detect occult genetic susceptibility to thrombophilia. Aim of the Work This study aims to detect Factor V Leiden (FVL), Prothrombin G20210A and MTHFR C677T Mutations among Patients with Budd-Chiari Syndrome. Patients and Methods This study was a cross sectional study conducted on 35 newly of Budd Chiari syndrome patients who were attending tropical department at Ain Shams University Hospitals during the period from April 2020 to January 2021. Informed consent was obtained from all patients in the study. Results results of our study revealed that JAK2 mutation it was detected in 17.1% of patients (6 patients), While PGM was detected in 2.9% of our patients. Furthermore Factor V Leiden Mutation was detected in 65.7%of patients and MTHFR mutation was detected in 48.6% of patients. The present study found no significant statistical difference between patients with Factor V Leiden Mutation & patients with normal Factor V Leiden regarding CBC parameters, levels of ANA, ACL IgM, ACL IgG, PC Deficiency, PS Deficiency and AT III Deficiency. In addition, our study revealed that there was no effect of MTHFR Mutation & JAK2 Mutation on levels of ANA, ACL IgM, ACL IgG, PC Deficiency, PS Deficiency and AT III Deficiency. Also there was no effect for MTHFR Mutation & JAK2 Mutation on outcome of patients, survival and response to treatment. Conclusion BCS is associated usually with genetic thrombophilia. Most commonly Factor V Leiden Mutation in 65.7%, followed by MTHFR mutation in 48.6% of our patients. Although, only Factor V Leiden Mutation has an effect on patients Outcome, Survival and Response to treatment through its effect on CBC parameters, levels of ANA, ACL IgM, ACL IgG, PC Deficiency, PS Deficiency and AT III Deficiency. Otherwise, JAK2 Mutation have significant effect on hemopoiesis.

Acute non-ST segment elevation myocardial infarction as the first manifestation of Takayasu arteritis in a 16-year-old female patient: a case report and literature review.

Takayasu arteritis (TA) is now recognized worldwide and is a disease that mainly affects the aorta and its main branches. TA rarely involves the small or medium-sized vessels. Certain vascular lesions, such as arterial stenosis, occlusion, and aneurysm are common with TA. However, patients with new-onset TA who present with left main trunk acute non-ST segment elevation myocardial infarction are extremely rare. We report a 16-year-old female patient with non-ST segment elevation myocardial infarction due to severe stenosis of the left main coronary artery that was caused by TA. She was eventually diagnosed with TA and underwent successful coronary artery stenting combined with glucocorticoids and folate reductase inhibitor therapy. Over the 1-year follow-up, she experienced two episodes of chest pain and was admitted to the hospital. During the second hospitalization, coronary angiography (CAG) revealed 90% stenosis of the original left main trunk (LM) stent. Following percutaneous coronary angiography (PTCA), drug-coated balloon (DCB) angioplasty was performed. Fortunately, a clear diagnosis of TA was made, and treatment was initiated with an interleukin-6 (IL-6) receptor inhibitor. Early diagnosis and therapy for TA are emphasized.

Effect of heparin on recurrent IVF-ET failure patients

Objective: To elucidate the possible role of unfractionated heparin in patients with failed repeated in in vitro fertilization and embryo transfer (IVF-ET) and thrombophilia. Methods: This case control study evaluated the efficacy of the unfractionated heparin in increasing the pregnancy and implantation ratio in women with recurrent IVF-ET failures. Eighty-six women received in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) with a record of three or more previous IVF-ET failures. Participants were randomly distributed into two groups. Group A (n=43) received unfractionated heparin 5 000 IU twice daily, and group B (n=43) did not take any antithrombotic drugs. Coagulation abnormalities such as factor V Leiden (FVL) mutation, methylene tetra hydro folate reductase (MTHFR) mutation and prothrombin mutation (FII) were evaluated. Age, body mass index, basal follicular stimulating hormone, basal estradiol, duration of infertility, and number of IVF-ET failures were compared between two groups. Results: 45.0% and 17.4% of women were pregnant with and without MTHFR and prothrombin mutation, respectively, when they received unfractionated heparin treatment. The implantation rate was more in group A (12.5%) than group B (4.3%) and differences in the fertilization rate of the two groups were observed (27.7% vs. 35.9%). The clinical pregnancy rate per cycle was remarkably more in group A (30.2%) than group B (14.0%). Conclusions: Heparin is a safe and valuable treatment for patients with repeated IVF-ET failures. The clinical pregnancy and implantation rates are higher in the heparin-treated group in contrast with the control group. Trial registration: The trial registration was done with clinical registration number of “ IRCT138807202575N1”.

Evaluation of homocysteine, folic acid and methylenetetrahydrofolate reductase C677T gene polymorphism associated with recurrent spontaneous miscarriage

This study was aimed to assess the association between evaluation of homocysteine level, folic acid and methylenetetrahydrofolate reductase MTHFR C677T gene polymorphism and recurrent pregnancy loss RSA. Sixty four women diagnosed with RSA and 41 normal fertile female, who had at least one or more child, 21 of them were pregnant women while 20 were non-pregnant. The level of homocysteine was high in women with RSA (11.41+3.75) (p<0.05) compared to normal fertile women either pregnant (7.02+2.13) or non-pregnant (6.84+2.08), low levels of folic acid was observed (6.35+2.09) in women with RSA (p<0.05) compared to fertile women either pregnant(10.08+3.08) or non-pregnant(11.27+3.15). No statistically significant difference was observed in the distribution of genotypes between cases and controls for MTHFR polymorphism. In conclusion, the present study reveals to lack of association of MTHFR C677T polymorphism in RSA women.

Pyridazinones: A versatile scaffold in the development of potential target‐based novel anticancer agents

AbstractPyridazinones are important nitrogen‐rich heterocyclic core that has been driving the substantial medicinal interest due to their wide range of biological activities. This privileged scaffold forms a central core in numerous potent compounds, which results in the development of novel anticancer drugs with fruitful biological activities. The current review article summarizes the progressive development of novel pyridazinone derivatives that are targets for numerous receptors such as C‐met kinase inhibitors, PARP inhibitors, Tubulin polymerization inhibitors, Dihydro folate reductase inhibitors, B‐Raf inhibitors, Bruton tyrosine kinase inhibitors, FER tyrosine kinase inhibitors, and fibroblast growth factors receptors. It features the various simple techniques for the synthesis of pyridazinones and also highlights the mechanistic insights into the anticancer properties of pyridazinone derivatives.

Detection of factor V G1691A, Prothrombin G20210A and Methylene tetra hydro folate reductase deficiency C677T Gene mutations among Sudanese Women with recurrent spontaneous abortion

The current study is a prospective analytical case control study designed to investigate the relationship between Factor V Leiden G1691A, methylenetetrahydrofolate reductase (MTHFR) C677T and to the prothrombin G20210A mutation variant and adverse pregnancy outcomes. Material and Method: The study included hundred Sudanese women who experienced three or more of the adverse pregnancy loss as case group during their reproductive in the Omdurman Maternity Hospital (Sudan) these compared with ninety-four control group healthy women with at least more than two normal pregnancies and without any history of adverse pregnancy outcome or recurrent miscarriages. The study group data collected using structure questionnaire which was used to collect information about age, parity, medical and obstetric history, smoking, family medical and obstetric history, residency and relative marriage. Blood samples were collected from participants and total genomic DNA was isolated from blood leukocytes and the frequency of these gene mutations in the patients and controls were determined using PCR-restriction fragment length. Results the mutation was detected in 8 out of 100 cases (8.0%) and in 6 out of 94 controls (6.4%) (P- Value > 0.05).

Association of MTHFR gene polymorphism C677T (rs1801133) studies with early primary knee osteoarthritis in a South Indian population: a hospital-based study.

Osteoarthritis (OA) is the most commonly occurring disease of middle and elderly population, which is characterized by focal loss of joint articular cartilage, osteophyte formation and sub chondral bone remodeling. Classical risk factors of OA include age, gender, weight, joint injury, trauma, however hereditary component is one of the main crucial factors. Several genome wide association studies and candidate gene approaches have identified genetic variants involved in the influence and association of OA. In the current study influence of Methylene tetra hydro folate reductase MTHFR C677T (rs1801133) gene with early primary knee OA was evaluated. In this study 400 samples were included (200 cases & 200 controls). DNA was extracted & processed for PCR- RFLP evaluation and genotype analysis. Statistical analysis was performed & results indicated a lack of association between MTHFR gene polymorphism and early primary KOA. The stratification was done based on age & gender and also both. Individual's i.e females below the age of 40 years are more prone to the disease when compared with males. MTHFR gene polymorphism showed a lack of association with early primary knee osteoarthritis. To the best of our knowledge this is the first study from south India.

MTHFR C677T and MTR A2756G Gene Polymorphism in Neural Tube Defect Patients and Its Association with Red Blood Cell Folate Level in Eastern Indian Population.

Single-nucleotide polymorphism (SNP) is a single-nucleotide change in a deoxyribose nucleic acid (DNA) sequence that occurs in >1% of population. Methylene tetra hydro folate reductase (MTHFR) C677T (rs1801133) and methionine synthase enzyme (MTR) A2756G (rs1805087) are two such SNPs occurring in coding sequence of the respective genes, which are frequently seen with neural tube defects (NTDs). MTHFR and MTR genes are involved in folate metabolism. The folate level in the course of pregnancy is treated as vital in the etiopathogenesis of NTDs. This study aims to explore the association of SNPs of both genes and red blood cell (RBC) folate levels in the predisposition to NTDs. The purpose of this investigation was to determine the relationship of NTDs with polymorphisms in MTHFR and MTR genotype and to estimate and compare the RBC folate levels in NTD patients and controls. A total of 397 individuals were enrolled (163 patients and 234 controls) for this observational study. Genotyping to find out MTHFR C677T and MTR A2756G was performed by polymerase chain reaction-restriction fragment length polymorphism technique from DNA extracted from the subject's blood. RBC folate level was estimated by chemiluminescence immunoassay method with the same blood sample. The total RBC folate levels were significantly less among cases compared to controls (P = 0.020). A significant difference for RBC folate was observed between case and control groups of various genotypes of MTHFR C677T, except heterozygote CT (P = 0.459). Among MTR A2756G, genotypes with only homozygous AA have significant difference (P = 0.003) for RBC folate levels. Among different types of NTDs, there were no significant differences for RBC folate levels. Among MTHFR C677T, T allele possessed 1.9 times risk compared to C allele for the occurrence of NTDs. In MTR A2756G polymorphism, the odds of developing NTDs were 1.6 times in heterozygous AG compared to homozygous AA. Similarly, the risk for NTDs was three times higher in subjects with both heterozygous AG and CT genotypes compared to wild-type homozygous AA and CC genotypes. The total RBC folate levels were significantly less among cases compared to controls, and the genotypes had no such effect in decrease in RBC folate levels. The presence of mutant allele in homozygous or heterozygous condition for both SNPs had increased risk associated with NTDs.

Patent Foramen Ovale Closure among Patients with Hypercoagulable States Maintained on Antithrombotic Therapy

Background: Percutaneous device closure was shown to effectively prevent recurrent strokes in patients with patent foramen ovale (PFO). Whether this protective effect is relevant for patients with hypercoagulable states (HCSs) is unknown as they were not represented in prior studies. Methods: Data on 136 consecutive patients with a PFO and clinically significant HCS were retrospectively collected. PFO closure and antithrombotic regimen were decided on an individual basis by the treating physicians, and adherence to therapy was routinely evaluated. The outcome was the occurrence of cerebrovascular accident (CVA) or transient ischemic attack (TIA). Results: HCS types consisted of antiphospholipid syndrome (31%), factor-5 Leiden mutation (22%), prothrombin mutation (18%), protein S deficiency (15%), protein C deficiency (7%), methyl-tetra-hydro folate reductase mutation (5%), and essential thrombocytosis (2%). 102 patients (75%) were maintained on anticoagulants and the remaining on antiplatelet therapy. PFO closure was undertaken in 85 (63%); antithrombotic therapy was not interrupted prior to or after the procedures. At a mean follow-up of 46 ± 8 months, 23 patients (17%; 95% confidence interval, 9.3–22%) experienced an outcome event, mainly in the form of CVAs (n = 15, 65%). In multivariable analysis, PFO closure was associated with a 5-fold decrease in the risk of CVA/TIA (p = 0.02). This effect was independent of the type of HCS or antithrombotic therapy. Conclusions: Among patients with HCSs maintained on anticoagulant or antiplatelet therapies, PFO closure was associated with a significantly lower risk of CVA or TIA.

A COMPARATIVE STUDY ON RISK OF OSTEOPOROSIS ASSOCIATED WITH THE USE OF VALPROATE VERSUS LEVETIRACETAM IN EPILEPTIC PATIENTS AND ITS RELATIONSHIP WITH METHYLENE TETRA-HYDRO FOLATE REDUCTASE GENOTYPES

Objective: The study objects at assessing and comparing the intensity of the effect of valproate (VPA) and levetiracetam (LV) on the bone mass in young adult epileptic patients while distinguishing their methylene tetra-hydro folate reductase (MTHFR) genotypes and correlating MTHFR polymorphism and antiepileptic drugs (AEDs) usage with the risk of development of osteoporosis.
 Methods: The study design was a comparative, prospective, and observational study. It was conducted at Princess Esra Hospital (PEH), Hyderabad and genotype testing was carried out at Salar-e-Millat Research lab (PEH). The consent was obtained from total 70 subjects, divided into three groups:
 Group 1: 18 patients receiving sodium VPA monotherapy
 Group 2: 17 patients receiving LV monotherapy
 Group 3: 35 healthy control subjects from general population.
 Patients of either gender within age group of 15–40 years, experiencing generalized tonic-clonic seizures or focal seizures, receiving the AED for duration of time ≥2 years were included in the study.
 Results: Our study showed significant correlation between the AEDs treatment and MTHFR polymorphism in predisposing osteoporosis.
 Conclusion: The variants of MTHFR gene (C677T) are prone to develop increased levels of homocysteine as a result of decreased activity of the enzyme in their bodies which are further increased in patients receiving AEDs. Monitoring of homocysteine levels in epileptic patients especially in the mutants of MTHFR gene along with their periodic testing of bone mineral density levels is recommended. Treatment for low folate and calcium levels is recommended in these patients to correct their deficiencies.

Potential role of folic acid in preventing male infertility associated with MTHFR gene C677T (rs1801133) polymorphism

Folic acid and methylenetetrahydrofolate reductase enzyme (MTHFR) gene C677T (rs1801133) SNP have been proofed to influence the spermatogenesis. This study aimed to investigate the biochemical, histological, and immunohistochemical (IHC) of adult male rabbits after induction of infertility by adenine, and the role of MTHFR C677T genotypes with or without folic acid therapy. Thirty adult male rabbits were divided into three groups: control (G1), adenine-induced infertility (G2), and adenine-induced infertility with folic acid therapy (G3). The PCR-RFLP was applied to detect the MTHFR C677T genotypes. Biochemical analyses of seminal fluid, homocysteine, and sex hormones were determined with histological and IHC examination of testicular tissue. Morphometric measurements and a TUNEL assay of seminiferous tubules were also performed. The results revealed a significant increase in homocysteine and sperm abnormalities, while sex hormones, sperm count and motility decreased significantly in the TT genotype compared with the CC genotype. Also, G2 displayed a significant increase in homocysteine and sperm abnormalities, with a significant decrease in sex hormones, sperm count and motility as compared to the G1 and G3. In conclusion, MTHFR 677C > T polymorphism is a genetic risk factor for male infertility and folic acid has a potential role in the treatment of male infertility.

Vitamin B17 Ameliorates Methotrexate-Induced Reproductive Toxicity, Oxidative Stress, and Testicular Injury in Male Rats.

Methotrexate (MTX; 4-amino-10-methylfolic acid) is a folic acid reductase inhibitor used to treat autoimmune diseases and certain types of cancer. Testicular toxicity resulting from MTX is a significant side effect that may cause subsequent infertility. The present study was conducted to examine the ameliorating effects of vitamin B17 (VitB17) against testicular toxicity induced by MTX in male rats. A total of 50 male albino rats were equally divided into five groups [control group; vitamin B17 group (VitB17) administered VitB17 only; methotrexate group administered MTX only; cotreated group, (VitB17+MTX) and posttreated group (MTX+VitB17)]. In methotrexate group (MTX), a significant decrease was observed in body weight and the testicular weight, as well as the levels of plasma testosterone, luteinizing hormone and follicle-stimulating hormone compared with control. The sperm count, viability, morphology index, total motility, and progressive motility also decreased in MTX rats compared with control. Furthermore, the levels of reduced glutathione, catalase, and superoxide dismutase, as well as proliferating cell nuclear antigen protein expression, in the testicular tissue decreased in MTX compared with control. In addition, MTX caused a significant increase in DNA and tissue damage compared with control. However, VitB17 ameliorated these effects, indicating that it has a preventative and curative effect against MTX-induced reproductive toxicity in male rats. The protective effect of VitB17 may be associated to its antioxidant properties as it possibly acts as a free-radical scavenger and lipid peroxidation inhibitor, as well as its protective effect on the levels of GSH, SOD, and CAT.

A study of hyperhomocysteinemia in cerebral venous sinus thrombosis.

Background & objectives:Vegetarianism may result in low vitamin B12 and acquired hyperhomocysteinemia leading to thrombotic conditions such as cerebral venous sinus thrombosis (CVST). The clinico-radiological presentation and outcome of patients with hyperhomocysteinemia may be different from those without, but there is a paucity of information. This study was undertaken to find out the relationship of homocysteine (Hcy) with vitamin B12, folic acid and methyltetrahydrofolate reductase (MTHFR) mutation in the patients with CVST, and compare clinico-radiological severity and outcome of patients with and without hyperhomocysteinemia.Methods:Ninety-six CVST patients in whom Hcy level was measured, were included, and their risk factors and neurological, magnetic resonance (MR) imaging and MR venography findings were noted. They were evaluated for prothrombotic conditions including Hcy, vitamin B12, folic acid and MTHFR 677C→T mutation. Three month outcome was categorized as death, poor and good.Results:Seventy three per cent patients had risk factors; hyperhomocysteinemia in 52.1 per cent, protein S deficiency in 47.8 per cent, protein C deficiency in 19.4 per cent, MTHFR 677C→T mutation in 30.7 per cent, antinuclear antibody 11 per cent, and Factor V Leiden mutation in two per cent each. Thirty two per cent patients with hyperhomocysteinemia had no other thrombotic cause, and 22 per cent of them had either vitamin B12 and or folic acid deficiency only. The patients with hyperhomocysteinemia more frequently had vitamin B12 deficiency (70 vs. 13%), MTHFR 677C→T mutation (47.5 vs. 9.1%) and superior sagittal sinus thrombosis (78 vs. 56.5%) than normal Hcy group. The clinico-radiological severity and outcome were similar.Interpretation & conclusions:Hyperhomocysteinemia was an important correctable risk factor of CVST in patients from northern India, and majority of them had either low vitamin B12 level or MTHFR mutation.

Association study of MTHFR-c677t with male infertility and reporting new potential SNPS/ sequence variants as a sourse of population genetic markers

It is estimated that about 15% of couples concern with the inability to conceive after 1 year of unprotected intercourse. Genetic and environmental factors play important roles in male infertility. MTHFR (Methylene Tetrahydro Folate Reductase) is encoded by a gene which maps to human chromosome 1p36.22. The c.677C>T mutation causes an amino - acid change of Alanine to Valine (Ala222Val) in exon 4 which is related to oligospermia and azoospermia. Our purpose was to investigate the association of rs1801133 in MTHFR gene with azoospermia and also genetic diversity of some Iranian folks. Peripheral blood samples of 100 men suffered from azoospermia and 55 controls collected from Infertility Clinic of Jihad-university of Qom. DNA was extracted by salting-out method and SNP-Genotyping was performed by PCR-RFLP technique and sequencing. Furthermore, statistical and population genetics analysis were done. The rs1801133 showed no significant relation with azoospermia according to statistical analysis. Furthermore, population genetics analysis showed that there is no genetic differentiation between Turk and Fars ethnics and case and control individuals. MTHFR studied SNP has no significant relation with azoospermia infertility, although further studies on more samples are demanded.

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.

P3566Ambulatory blood pressure response to riboflavin supplementation in adults with the C677T polymorphism in the folate metabolising enzyme methylenetetrahydrofolate reductase (MTHFR)

Abstract Background Epidemiological evidence suggests that the C677T polymorphism in the folate metabolising enzyme MTHFR is associated with a 24–87% increased risk of hypertension. Riboflavin (a cofactor for MTHFR) can modify this phenotype, as demonstrated in a series of RCTs, at this centre. ABPM measures BP over 24hrs and provides a more reliable assessment of an individual's BP compared to clinic BP. Aim To investigate the effect of riboflavin supplementation on BP (by ABPM) in adults with the MTHFR 677TT genotype. Methods Adults (n=3462) were recruited and screened for the MTHFRTT genotype and those eligible (n=81), were stratified by baseline SBP and randomised to receive riboflavin (10mg/day/16weeks) or placebo. Riboflavin status was measured using the erythrocyte glutathione reductase activation coefficient (EGRac) assay and clinic and ABPM were measured in accordance with clinical guidelines. Results Mean 24hr, daytime and night-time SBP was 6mmHg higher (p<0.05) in adults with the TT v CC genotype. BP response to riboflavin was strongly dependent on baseline BP. In participants with a baseline SBP≥125mmHg, riboflavin resulted in a significant BP lowering in daytime SBP (Table). Day-time SBP response to riboflavin Treatment Placebo P-value Age 48.6 (11.3) 47.8 (12.0) 0.823 Male n (%) 17 (73.9) 12 (46.2) 0.093 BMI 28.8 (3.8) 27.2 (3.1) 0.131 Riboflavin status (EGRac)† Pre 1.33 (1.28–1.38) 1.29 (1.24–1.34) Post 1.20 (1.14–1.25) 1.31 (1.25–1.36) <0.001 Change −0.12 0.02 Daytime BP (mmHg)‡ Pre 137.1 (132.7–141.3) 134.7 (130.3–139.1) Post 133.0 (128.9–138.1) 134.8 (130.6–139.1) 0.036 Change −3.8 −0.2 ANCOVA, adjusting for sex. †Higher EGRac values indicate lower riboflavin status; ‡mean of participant daytime/awake hrs personalised for each participant. Conclusion This is the first study to show, using ABPM, that riboflavin supplementation, targeted at adults with the MTHFR 677TT genotype, results in significant lowering of mean, day and night BP. Given the frequency of this genotype worldwide (approximately 10%, but as high as 30% in some populations) these findings could offer a personalised approach for BP management in at-risk sub-populations. Acknowledgement/Funding This work was supported by DSM Nutritional Products, Ltd and Wellcome Trust-Wolfston Northern Ireland Clinical Research Facility are acknowledged

The Role of Polymorphism Gen Methylene Tetra Hydrofolate Reductase (MTHFR) C677T in Ischaemic Stroke Patients with and Without Hypertension.

BACKGROUND:Stroke is a leading cause of disability and remains the second leading cause of death in the world. Some of the pathogenesis of stroke are interactions between genetic and acquired risk factors, the interaction is related with the atherosclerotic which is the main pathogenesis of ischaemic stroke. Previous studies demonstrated an association between methylene tetra hydro folate reductase (MTHFR) genotype and ischaemic stroke; the MTHFR C677T genotype is one of the independent risk factor.AIM:This study aims to know about the role of polymorphism gen MTHFR C677T in ischaemic stroke patients with and without hypertension.METHODS:This study is a cross-sectional study, the sample was taken consecutively, after approval by the Medical Faculty Science’s Ethics Committee at University Sumatera Utara. All sample matched with inclusion and exclusion criteria, demography data and blood sample were taken. Demography data were analysed using descriptive statistic.RESULTS:Of the 106 ischaemic stroke patients were divided into two groups, the first group is patients with hypertension (53 patients), and the second group is without hypertension (53 patients). We have done the PCR- RFLP to all the patients, we got 78 patients with 677CC of MTHFR genotype, 23 patients with 677CT genotype and 5 patients with 677TT genotype. We found polymorphism C677T is more frequent in ischaemic stroke patients with hypertension (16 patients; 69.5%), and all the patient with 677TT genotype are an ischaemic stroke with hypertension (5 patients; 100%).CONCLUSION:We concluded that polymorphism MTHFR C677T have an important role in hypertension and ischaemic stroke.