Methylene tetrahydrofolate reductase (MTHFR) and vascular endothelial growth factor (VEGF) polymorphisms in Brazilian patients with Hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC)

OBJECTIVE: The folate pathway is involved in hepatic carcinogenesis and angiogenesis. Polymorphisms in genes related to such processes, including methylene tetrahydrofolate reductase (MTHFR) and vascular endothelial growth factor (VEGF)] may play an important role in the development of hepatocellular carcinoma (HCC). The objective of this study was to evaluate MTHFR and VEGF polymorphisms in Brazilian patients with hepatitis C virus (HCV)-related HCC. METHODS: A total of 119 patients diagnosed with confirmed HCC and HCV were included in the study. SNP genotyping assays were performed using real-time PCR. VEGFA (rs2010963, rs3025039, and rs833061) and MTHFRC677T (rs1801133, rs1801131) polymorphisms were evaluated. RESULTS: The C alleles of MTHFR (rs1801131) and VEGF (rs2010963) were associated with protection against the development of multinodular HCC, while the T allele of MTHFR (rs1801133) was associated with a higher risk of multinodular presentation [p=0.04 OR 1.835 CI (1.022-3.297)]. Multivariate analysis revealed that the GG/GC genotypes of VEGF rs2010963 were independently associated with multinodular tumors at diagnosis (p=0.013; OR 4.78 CI (1.38-16.67)]. CONCLUSION: Our results suggest that these polymorphisms may increase the risk of rapid tumor progression in patients with HCV infection. This subgroup of patients with HCC and who present polymorphism is more likely to be diagnosed with multinodular disease and not be amenable to receiving curative treatments. These data must be validated in larger cohorts, and the screening intervals can be customized based on genetic history.

Thromboelastogram parameters and MTHFR gene polymorphism in the diagnosis of adverse fetal pregnancy outcomes / 中国输血杂志

【Objective】 To retrospectively analyze the situation of patients with adverse fetal outcomes by thromboelastogram (TEG) parameters and, MTHFR gene polymorphism, so as to provide molecular biological diagnosis basis for patients with adverse pregnancy outcomes, and a new scheme for early prevention and treatment of women of childbearing age with MTHFR gene polymorphism. 【Methods】 A total of 100 women with adverse fetal pregnancy outcomes were selected as the adverse pregnancy group, and 100 healthy women of childbearing age with normal pregnancy history were selected as the controls. MTHFR gene C677T and A1298C polymorphisms were detected by polymerase chain reaction (PCR). TEG and blood coagulation were detected in the experimental group. 【Results】 The A1298C gene polymorphism(AA、CC、AC; A、C) was similar in both adverse pregnancy group and the controls. The frequency distribution of C, T allele of MTHFR gene C677T was statistically significant (χ2=4.60, P<0.05, OR =1.645, 95% CI: 1.042~2.595). TT and CT+ CC types showed significant different association with the factors of stillbirth(χ2 =7.49, P<0.05). MA value of TEG in the diagnosis of TT type of C677T genotypes MTHFR in 32 patients with adverse pregnancy outcome was analyzed. The area under the AUC curve of MA value was 0.795. 【Conclusion】 MTHFR C677T polymorphism TT with TEG parameter hypercoagulability is an important risk factor in the occurrence of pregnancy stillbirth in adverse pregnancy outcomes.

Evaluation of Relationship of the Methylene Tetrahydrofolate Reductase Enzyme Polymorphisms with Serum Methotrexate Concentration and Toxicity in Acute Lymphoblastic Leukemia Patients Treated with High Dose Methotrexate Infusion

Background: Methotrexate (MTX) blocks MethyleneTetrahydrofolate Reductase (MTHFR) Enzymethereby, interrupt folate metabolism, it is used in thetreatment of cancer and autoimmune disorders. Aimand Objectives: The present study aimed to evaluatethe relationship of the MTHFR polymorphisms withserum MTX concentration and its toxicity in AcuteLymphoblastic Leukemia (ALL) patients treated withhigh dose MTX infusion. Material and Methods: Levelof Serum MTX was measured, along with the detectionof MTHFR polymorphisms viz. C677T and A1298Cby Polymerase Chain Reaction (PCR) followed byDNA sequencing. The percentages of toxicitydeveloped in patients were calculated among the wildtype and carriers for both polymorphisms and werecompared between the groups. Results:The majority ofpatients 36 (72 %) were wild type for the C677Tpolymorphism and 32 (64 %) of patients were carriersfor the A1298C polymorphism [48% heterozygous(AC), and 16 % homozygous (CC)]. Among 50 ALLpatients studied, significant difference was noted in thegenotype and allele frequencies for C677Tpolymorphism, while only allele frequencies differedsignificantly for A1298C polymorphism. The serumMTX level at 48 hours after the start of High DoseMTX (HDMTX) infusion of the C677T variant (CT)was slightly high in all four cycles however, in the firstcycle, there was a significant increase in the level ofMTX. There was no significant difference in the serumMTX level found in all four cycles between patientswild type and carriers for A1298C polymorphism. ForA1298C polymorphism, the mean SGPT level incarriers was significantly high as compared to wildtype. Conclusion: The present study concludes thatpatients with C667T variant had elevated serum MTXconcentration at 48 hours after the start of HDMTXinfusion.

In silico analysis of the structural and functional implications of SLC19A1 R27H polymorphism

In view of the documented association of solute carrier family 19 member 1 (SLC19A1) G80A (R27H) polymorphism with the risk for different types of cancers and systemic lupus erythematosus (SLE), we have reanalysed the case–control study on breast cancer to ascertain the conditions in which this polymorphic variant exerts the risk of breast cancer. Association statistics have revealed that this polymorphism exerts the risk for breast cancer under the conditions of low folate intake, and in the absence of well-documented protective polymorphism in cytosolic serine hydroxymethyltransferase. To substantiate this observation, we have developed a homology model of SLC19A1 using glycerol-3-phosphate transporter (d1pw4a) as a template where 73% of the residues were modelled at 90% confidence while 162 residues were modelled ab initio. The wild and mutant proteins shared same topology in S3, S5, S6, S7, S11 and S12 transmembrane domains. The topology varied at S1 (28–43 residue vs 28–44 residue), S2 (66–87 residue vs 69–87 residue), S4 (117–140 residue vs 117–139 residue), S8 (305–325 residue vs 305–324 residue), S9 (336–356 residue vs 336–355residue), and S10 (361–386 residue vs 361–385 residue) transmembrane domains between wild versus mutant proteins. S2 domain is shortened by three amino acid residues in themutantwhile in other domains the difference corresponds to one amino acid residue. The 3DLigandSite analysis revealed that the metallic-ligand-binding sites at 273Trp, 277Asn, 379Leu, 439Phe and 442Leu are although unaffected, there is a loss of active sites corresponding to nonmetallic ligand binding. Tetrahydrofolate and methotrexate have lesseraffinity towards the mutant protein than the wild protein. To conclude, the R27H polymorphism affects the secondary and tertiary structures of SLC19A1 with the significant loss in ligand-binding sites.

Association of genetic polymorphis of thymidylate synthase and methylene tetrahydro folate reductase with therapeutic effect of pemetrexed for treating advanced lung adenocarcinoma patients / 实用医学杂志

Objective To investigate the expressions of thymidylate synthase (TS) and methylene tetrahydrofolate reductase (MTHFR) polymorphisms and the therapeutic efficacy of chemotherapy with pemetrexed and platinum in advanced lung adenocarcinoma patients. Methods Fifty-eight patients with advanced lung adenocarcinoma were enrolled in this study. The blood samples from 25 of them were examined for extraction of DNA. The associations of the gene polymorphisms with the chemotherapy efficacy and PFS were analyzed. Results Disease control rate was noted by 38％ and the median time of progression-free survival was 8.1 months among 58 patients.There were 16％, 32％, 52％, and TS genotypes as 2R/2R, 2R/3R and 3R/3R respectively; the difference in the control rate between those with TS gene of 3R/3R and those with TS gene of R/2R+2R/3 R was significant statistically (53.8％ vs. 91.7％, P = 0.046) , but the difference in PFS was statistically insignificant (9.3 vs. 10.4 months, P> 0.05). There were 40％, 52％, 8％, and MTHFR genotypes as CC, CT and TT respectively. The DCR in those with MTHFR CC and C/T + T/T was 70％ and 73.3％, respectively and PFS was 10 months and 9.7 months respectively, showing no significant difference (P> 0.05). Conclusion The TS gene polymorphism is associated with therapeutic effect of pemetrexed for advanced lung adenocarcinoma, but MTHFR is not.

Rapid detection of human MTHFR gene polymorphism by the TaqMan-ARMS method / 临床检验杂志

Objective To establish a rapid detection method for human methylene tetrahydrofolate reductase ( MTHFR) gene polymor-phism by using the primer mismatching amplification and fluorescence quantitative PCR. Methods A total of 214 samples with differ-ent MTHFR C677T genotypes ( CC, CT, TT) or different A1298C genotypes ( AA, AC, CC) , which were verified by gene sequen-cing, were collected, and the plasmids with the corresponding wild-type and homozygous mutants were constructed, respectively. The amplification refractory mutation system ( ARMS) primers and TaqMan probes were designed based on the wild-type standard sequence of MTHFR gene, and the optimal mutation detection system was established. The results from the system were compared with the known sequencing results to verify the feasibility of the system. Results The performance of the established TaqMan-ARMS method was ex-cellent, which had 10 copies/μL of lowest detectable limit and high specificity. There was no nucleic acid amplification in the cross de-tection between samples and the negative control. In addition, the established method had good repeatability. The standard deviations of the reproducibility detection of MTHFR-667 and 1298 loci ranged from 0.11 to 0.44, and the coefficients of variation ( CV) of homozy-gous and heterozygous samples were all less than 4.52％. The consistency of the established method with the sequencing method in 214 clinical samples was 100％. Conclusion The established TaqMan-ARMS method for the detection of MTHFR gene polymorphism is simple, rapid and accurate, which may be used for the rapid diagnosis of clinical patients.

Efecto del etiquetado calórico sobre la elección de un menú: estudio preliminar en Santiago de Chile / Effect of calorie labeling on menu selection: a preliminary study in Santiago, Chile

Folate is an essential nutrient because mammals lack biological activity to synthesize. It different factors generate folate deficiency. Recent studies have identified that the C677T variant of the enzyme methylene tetrahydrofolate reductase (MTHFR), can play a role in serum folate concentrations (SFC) and red cell folate (RCF). The aim of this rewiev was to actualice some generalities of folate metabolism, factors related to its deficiency, biochemical indicators used to assess the nutritional status of folate and role of the C677T polymorphism of the MTHFR enzyme on the cycle of folate and methionine. It is necessary to design studies with representative samples corroborating the effect of polymorphisms on biochemical indicators of nutritional status of folate and determine the dose-response effect and contribute to modify the nutritional recommendations with the necessary scientific evidence.

El 62% de la población chilena presenta sobrepeso (dato OMS). Publicar calorías en menús de restaurantes podría ayudar a controlar este problema. El objetivo fue estudiar el efecto de la entrega de información calórica en la elección de almuerzos típicos. La metodología tuvo un enfoque cuantitativo, con encuesta on line de diseño transversal, estructurada, con preguntas abiertas y cerradas, y con escala tipo Lykert. Se obtuvo 227 respuestas válidas. Los encuestados seleccionaron un almuerzo de 3 elementos, antes y después de exponer información calórica. Los resultados mostraron que el 49% de los encuestados reduce en promedio 292 kcal, (39,2% del total de calorías en menú) al considerar la información calórica. Nuestros resultados sugieren que aproximadamente para la mitad de los encuestados, la información fue útil en la selección de menú. Esta información podría ayudar a profesionales de la Salud a crear conciencia, facilitando a los consumidores elecciones más saludables.

MTHFR C677T and A1298C Gene Polymorphisms and Ischemic Heart Disease (IHD) in a Sri Lankan Population - A Preliminary Study

MTHFR A1298C and C677T SNPs are now recognised as important genetic mutations which would give rise to hyperhomocysteinemia. In this study, we analysed the prevalence of these two SNPs in 79 ischemic heart disease (IHD) patients awaiting coronary artery bypass grafting and 79 healthy subjects. MHFR polymorphisms were analysed using polymerase chain reaction followed by a restriction fragment analysis. Prevalence rates for MTHFR C677T polymorphism were 72.8%, 24.7%, and 2.5% for CC, CT, and TT genotypes, respectively, for the whole study population with 677CC genotype being the predominant genotype among both the IHD patients and the controls. The 677TT genotype was detected only among the IHD patients. There was no significant difference in MTHFR 677 genotype variations between IHD patients and the control group. Prevalence rates for the MTHFR A1298C polymorphism were 50%, 37.3%, and 12.7% for the AA, AC, and CC genotypes, respectively, for the whole study population with 1298AA genotype being the predominant genotype among controls and 1298AC the predominant genotype among IHD patients. There was a significant difference (p < 0.01) between IHD patients and controls when the MTHFR 1298 genotype variations were compared. Allele frequencies for the mutant T allele for C677T mutation at 0.149 are the highest reported from Sri Lanka. The frequency of the C for the A1298C mutation was 0.313. Results of this study indicate that MTHFR A1298C SNP is more prevalent in Sri Lankans when compared to C677T SNP and that the mutant forms of the MTHFR A1298C SNP are associated with ischemic heart disease.

Analysis of the Efficacy of Neoadjuvant Chemotherapy in Advanced Gastric Carcinoma Patients before Laparoscopic Minimally Invasive Resection / 中国药房

OBJECTIVE:To observe the efficacy and safety of neoadjuvant chemotherapy in advanced gastric carcinoma patients before laparoscopic minimally invasive resction.METHODS:In retrospective analysis,93 patients with advanced gastric carcinoma were divided into single group (55 cases) and combined group (38 cases).Single group received laparoscopic minimally invasive operation.Combination group was given Tetrahydrofolate injection 400 mg/m2+Oxaliplatin injection 85 mg/m2,i.v.,2 h,d1+ Fluorouracil injection 2 400 mg/m2,i.v.,46 h,d2.A treatment course lasted for 2 weeks,both received 2-4 courses of treatment and 4 weeks later received laparoscopic minimally invasive resection.Both groups received routine treatment as parenteral nutrition and preventive anti-inflammation.They were given oxaliplatin+capecitabine or gimeracil oteracil potassium capsule+oxaliplatin chemotherapy for 6 courses.Clinical efficacies and ADR of combination group were observed.Operation time,intraoperative blood loss,intraoperative blood transfusion,resection range,conversions to laparotomy,the number of lymph node dissection,complete resection and postoperative first exhaust time,the time of fluid feeding recovery,the length of hospital stay and complications were observed in 2 groups.RESULTS:The objective remission rate and disease control rate of combination group were 44.8％ and 92.2％;there were 23 case time of grade Ⅰ ADR,13 case time of grade Ⅱ ADR and 3 case time of grade Ⅲ ADR.Complete resection rate of combination group was significantly higher than that of single group,with statistical significance (P＜0.05).There was no statistical significance in operation time,intraoperative blood loss,intraoperative blood transfusion,resection range,conversions to laparotomy,the number of lymph node dissection,postoperative first exhaust time,the time of fluid feeding recovery,the length of hospital stay or the incidence of complications between 2 groups (P＞0.05).CONCLUSIONS:For advanced gastric carcinoma patients before laparosapic minimally invasive resection,tetrahydrofolate,oxaliplatin and fluorouracil neoadjuvant chemotherapy show therapeutic efficacy and can improve the rate of complete resection without increasing the incidence of ADR.

Relationship between Polymorphism of MTHFR and Curative Effect of Adjuvant Chemotherapy in Postoperative Colorectal Cancer Patients / 中国药学杂志

OBJECTIVE: To investigate the association between methylene tetrahydrofolate reductase (MTHFR)C677T polymorphism and the efficacy of the adjuvant chemotherapy with XELOX regimen for patients who had underwent radical resection of colorectal cancer. METHODS: Sixty-two patients who had received chemotherapy with XELOX regimen following radical resection of colorectal cancer were tested for MTHFR C677T polymorphism using Kompetitive Allele-Specific PCR to analyze the association of MTHFR C677T polymorphism with the prognosis and adverse reactions to chemotherapy. RESULTS: Among the 62 patients with colorectal cancer, there were 3 allelotypes (C/C, C/T and T/T)at the MTHFR C677T locus, and their frequencies were 46.8%, 40.3%, and 12.9%, respectively. The recurrence free survival time was prolonged in C/T and T/T group than C/C group(Log-rank=4.778, P0.05). CONCLUSION: MTHFR C677T polymorphism is associated with the prognosis with adjuvant chemotherapy with XELOX regimen, and is not associated with the toxicities of chemotherapy. TNM stage IV is predicative of worse prognosis with postoperative adjuvant chemotherapy.

Association between MTHFR polymorphisms and colorectal cancer / 国际肿瘤学杂志

Colorectal cancer (CRC) is one of the most common malignancies in the world.Methylene tetrahydrofolate reductase (MTHFR) is a key enzyme of folate metabolism process.In recent years,many studies indicate that MTHFR polymorphisms are significantly associated with the morbidity,response to chemotherapy and prognosis of CRC,but conclusions are inconsistent and remain to be further confirmed.

Association between MTHFR polymorphisms and colorectal cancer / 国际肿瘤学杂志

Colorectal cancer (CRC) is one of the most common malignancies in the world.Methylene tetrahydrofolate reductase (MTHFR) is a key enzyme of folate metabolism process.In recent years,many studies indicate that MTHFR polymorphisms are significantly associated with the morbidity,response to chemotherapy and prognosis of CRC,but conclusions are inconsistent and remain to be further confirmed.

Analysis of the Efficacy of Neoadjuvant Chemotherapy in Advanced Gastric Carcinoma Patients before Laparoscopic Minimally Invasive Resection / 中国药房

OBJECTIVE:To observe the efficacy and safety of neoadjuvant chemotherapy in advanced gastric carcinoma patients before laparoscopic minimally invasive resction.METHODS:In retrospective analysis,93 patients with advanced gastric carcinoma were divided into single group (55 cases) and combined group (38 cases).Single group received laparoscopic minimally invasive operation.Combination group was given Tetrahydrofolate injection 400 mg/m2+Oxaliplatin injection 85 mg/m2,i.v.,2 h,d1+ Fluorouracil injection 2 400 mg/m2,i.v.,46 h,d2.A treatment course lasted for 2 weeks,both received 2-4 courses of treatment and 4 weeks later received laparoscopic minimally invasive resection.Both groups received routine treatment as parenteral nutrition and preventive anti-inflammation.They were given oxaliplatin+capecitabine or gimeracil oteracil potassium capsule+oxaliplatin chemotherapy for 6 courses.Clinical efficacies and ADR of combination group were observed.Operation time,intraoperative blood loss,intraoperative blood transfusion,resection range,conversions to laparotomy,the number of lymph node dissection,complete resection and postoperative first exhaust time,the time of fluid feeding recovery,the length of hospital stay and complications were observed in 2 groups.RESULTS:The objective remission rate and disease control rate of combination group were 44.8％ and 92.2％;there were 23 case time of grade Ⅰ ADR,13 case time of grade Ⅱ ADR and 3 case time of grade Ⅲ ADR.Complete resection rate of combination group was significantly higher than that of single group,with statistical significance (P＜0.05).There was no statistical significance in operation time,intraoperative blood loss,intraoperative blood transfusion,resection range,conversions to laparotomy,the number of lymph node dissection,postoperative first exhaust time,the time of fluid feeding recovery,the length of hospital stay or the incidence of complications between 2 groups (P＞0.05).CONCLUSIONS:For advanced gastric carcinoma patients before laparosapic minimally invasive resection,tetrahydrofolate,oxaliplatin and fluorouracil neoadjuvant chemotherapy show therapeutic efficacy and can improve the rate of complete resection without increasing the incidence of ADR.

Factores determinantes del estado nutricional del folato y el rol de la variante genética C677T de la enzima metilen tetrahidrofolato reductasa (MTHFR) / Determinants of folate nutritional status and the role of the genetic variation of C677T methylene tetrahydrofolate reductase (MTHFR) enzyme

Folate is an essential nutrient because mammals lack biological activity to synthesize. It different factors generate folate deficiency. Recent studies have identified that the C677T variant of the enzyme methylene tetrahydrofolate reductase (MTHFR), can play a role in serum folate concentrations (SFC) and red cell folate (RCF). The aim of this rewiev was to actualice some generalities of folate metabolism, factors related to its deficiency biochemical indicators used to assess the nutritional status of folate and role of the C677T polymorphism of the MTHFR enzyme on the cycle of folate and methionine. It is necessary to design studies with representative samples corroborating the effect of polymorphisms on biochemical indicators of nutritional status of folate and determine the dose-response effect and contribute to modify the nutritional recommendations with the necessary scientific evidence.

El folato es un nutriente esencial porque los mamíferos carecen de actividad biológica para sintetizarlo. Diferentes factores generar deficiencia de folato. Estudios recientes han identificado que la variante C677T de la enzima metilen tetrahidrofolato reductasa (MTHFR), puede jugar un papel en las concentraciones de folato sérico (FS) y eritrocitario (FE). El objetivo de este trabajo fue revisar algunas generalidades del folato, su metabolismo, los factores relacionados con su deficiencia, los indicadores bioquímicos utilizados para evaluar el estado nutricional del folato y el papel del polimorfismo C677T de la enzima MTHFR sobre el ciclo del folato y de la metionina. Es necesario diseñar estudios con muestras representativas que corroboren el efecto de los polimorfismos sobre los indicadores bioquímicos del estado nutricional del folato y determinar el efecto dosis-respuesta y así contribuir con la evidencia científica necesaria para modificar las recomendaciones nutricionales.

Methylene tetrahydrofolate reductasel reductase C677T polymorphisms and esophageal cancer susceptibility: A meta‑analysis based on Chinese Han population.

OBJECTIVE: The aim of this meta‑analysis was to assess the methylene tetrahydrofolate reductase (MTHFR) gene C677T polymorphisms and esophageal cancer susceptibility in Chinese Han population. MATERIALS AND METHODS: The databases of PubMed, MEDLINE, Wanfang, and CNIK was electronic searched to find the case–control or cohort study about the relationship between MTHFR gene C677T polymorphisms and esophageal cancer susceptibility in Chinese Han population. The odds ratio (OR) was used to assess the relationship between CC, CT, and TT genotype and esophageal cancer risk. The data were pooled using Stata 11.0 software. RESULTS: Eight articles included 1752 esophageal cancer and 2363 controls were found and included in this meta‑analysis. The pooled OR was 1.86 with its 95% confidence interval of 1.21–2.86 and 1.62 with its 95% confidence interval of 1.15–2.27 for TT versus CC and CT versus CC model which indicated that people with TT OR CT genotype significant increase the risk of developing esophageal cancer. CONCLUSION: Esophageal cancer risk was significantly increased in people with TT/CT genotype of MTHFR gene.

Effect of fluorouracil for injection on methylenetetrahydrofolate reductase, glutathione S-transferase and tumor markers on patients with advanced gastric cancer / 中国生化药物杂志

Objective To investigate the effect of fluorouracil for injection on methylene tetrahydrofolate reductase ( MTHFR ) , glutathione S-transferase ( GST) and tumor markers in patients with advanced gastric cancer.Methods Forty-eight cases with advanced gastric cancer were selected from August 2013 to August 2014 in the hospital and divided into two groups.The control group (n=24) were treated with capecitabine plan, and the experiment group ( n=24 ) were treated with fluorouracil for injection plan.MTHFR concentration, GST activity and tumor markers [ carbohydrate antigen-199(CA199), carcinoembryonic antigen (CEA), alpha fetoprotein(AFP)] levels were compared before and after treatment.ResuIts Compared with control group, MTHFR concentration of experiment group was higher (P<0.05), GST activity was lower (P<0.05), the tumor markers levels were lower (P<0.05), total effective rate of clinical symptoms was higher [16(66.67) vs.22(91.67), χ2 =4.55, P<0.05], and incidence of adverse reactions was lower [14(58.33) vs.6(25.00), χ2 =5.49, P<0.05].ConcIusion Fluorouracil for injection chemotherapy could increase the serum MTHFR concentration, reduce GST activity, and its clinical curative effect is obvious with less adverse reactions, which has important significance in treatment of patients with advanced gastric cancer.

Influnce of MTHFR gene polymorphism on folicacid in lowering plasma homocyteine / 实用医学杂志

Objective To investigate the impact of methylene tetrahydrofolate reductase (MTHFR) gene polymorphism on folicacid for lowering plasma level of homocysteine in elderly patients with coronary heart disease (CHD). Methods In the first affiliated hospital of Zhengzhou university,a total of 180 elderly patients with CHD were randomized to two groups. The study group (91 ptients) received folicacid 5mg once daily and the control group (89 patients) received no folic acid. 8 weeks after treatment, the changes in plasma Hcy were observed and analyzed by MTHFR genotypes (TT vs. CC vs. CT). Results Plasma Hcy of the three genotypes had a statistical significance at the baseline (P<0.001), among which plasma Hcy level was the hightest in patients with genotype TT, while it had on difference in genotypes CC and CT (P = 0.057). 8 weeks after treatment, plasma Hcy level declined up to 24%in the patients with genotype TT in the study group, while it decreased about 6%and 15%in patients with genotype CC or CT, separately. Hcy level was slightly decreased in the patients with genotype CC, CT, or TT in the control group. Conclusions Plasma Hcy level differs statistically in three genotypes, and it is the highest in genotyp TT. Folicacid can effectively lower the level of plasma Hcy in elderly patients with CHD, especially those with genotype TT.

Biochemical characterization of the bifunctional enzyme dihydrofolate reductase-thymidylate synthase from Leishmania ( Viannia ) and its evaluation as a drug target / Caracterización bioquímica de la enzima bifuncional dihidrofolato reductasa-timidilato sintasa de Leishmania ( Viannia ) y su evaluación como blanco molecular

Introduction. Dihydrofolate reductase (DHFR) has been used successfully as a drug target in the area of anti-bacterial, anti-cancer and anti-malarial therapy. Although this bifunctional enzyme is also a potential drug target for treatment of leishmaniasis, there have been no reports on its efficacy against Leishmania ( Viannia ) species . Materials and methods. The gene encoding the bifunctional DHFR and thymidylate synthase (TS) of Le. (V.) braziliensis was isolated and expressed in E. coli. The enzyme was purified and characterized. The inhibitory effects of antifolates and four aporphine alkaloids on its activity were evaluated. Results. The full-length gene consists of a 1560-bp open reading frame encoding a 58 kDa translated peptide containing DHFR and TS domains linked together in a single polypeptide chain. The recombinant DHFR-TS enzyme revealed K m and V max values of 55.35 ± 4.02 µ M (mean ± SE) and 0.02 ± 5.34 x 10 -4 µ M/min respectively for dihydrofolic acid (H 2 F). The Le. braziliensis rDHFR-TS have Ki values for antimicrobial antifolates in the µM range. Methotrexate (MTX) was a more-potent inhibitor of enzymatic activity ( Ki = 22.0 µM) than trimethoprim ( Ki = 33 µM) and pyrimethamine ( Ki = 68 µM). These Ki values are significantly lower than those obtained for the aporphine alkaloids. Conclusion. The results of the study show the inhibitory effect of antifolate drugs on enzymatic activity, indicating that Le. braziliensis rDHFR-TS could be a model to studying antifolate compounds as potential antiprotozoal drugs.

Introducción. La dihidrofolato reductasa (DHFR) se ha utilizado como blanco molecular en tratamientos antibacterianos, anticancerígenos y antipalúdicos. También, actúa como blanco molecular en Leishmania ; sin embargo, no existen reportes de la enzima bifuncional en especies de Leishmania ( Viannia ). Materiales y métodos. Se ha aislado y expresado en Escherichia coli el gen que codifica para la enzima bifuncional DHFR y la timidilato-sintasa (TS) de Leishmania braziliensis . La enzima recombinante se purificó y caracterizó, y se evaluó el efecto inhibitorio de algunos antifolatos, así como de cuatro alcaloides aporfínicos. Resultados. El gen se compone de aproximadamente 1.560 pb y codifica un péptido de 58 kDa que contiene los dominios DHFR y TS ligados en una sola cadena polipeptídica. La enzima recombinante DHFR-TS, utilizando el dihidrofolato (H2F) como sustrato, presentó valores de K m y V max de 55,35 ± 4,02 (media ± el error estándar de la media) y de 0,02 ± 5,34 x 10 -4 , respectivamente. La enzima rDHFR-TS de L. braziliensis presentó valores de Ki para los antifolatos en el rango de micras. El metotrexato fue el inhibidor más potente de la actividad enzimática ( Ki =22,0 mM) en comparación del trimetoprim ( Ki =33 mM) y la pirimetamina ( Ki =68 mM). Estos valores de Ki son significativamente más bajos en comparación con los obtenidos para los alcaloides aporfínicos. Conclusión. Los resultados muestran el efecto inhibitorio de los antifolatos sobre la actividad enzimática, lo cual indica que la rDHFR-TS de L. braziliensis podría ser un modelo para estudiar moléculas antiprotozoarias potenciales.

Livedoid Vasculopathy with Hyperhomocysteinemia due to MTHFR Mutation / 대한피부과학회지

Livedoid vasculopathy is a hyalinizing vascular disease characterized by thrombosis and ulceration of the lower extremities. It can be caused by an alteration in control of coagulation with the formation of thrombi within dermal blood vessels. We report a case of livedoid vasculopathy with hyperhomocysteinemia due to MTHFR mutation, which is treated by folic acid and which also showed very unusual clinical manifestations. A 38-year-old male visited the department of dermatology with a 1 year history of purplish-brown purpura with punched-out ulcers on both lower legs. He had a history of homocysteinemia due to methylene tetrahydrofolate reductase (MTHFR) mutation. The histopathologic findings of the lesional skin revealed dense superficial and deep perivascular and perifollicular infiltrates of lymphocytes and fibrin deposition within the vessels in the dermis. On the basis of clinical and pathological findings, livedoid vasculopathy with hyperhomocysteinemia due to MTHFR mutation was diagnosed and improved by the treatment of 1 mg of folic acid daily.

High-throughput virtual screening for novel dhfr inhibitors of P. Jiroveci from the zinc database.

Many of the opportunistic infections that occur at this late stage can be fatal and since that Pneumocystis carinii pneumonia (PCP) is a leading opportunistic infection found among immunocompromised (CD4 cell < 200) patients worldwide. DHFR is responsible for the growth and maturation of sporozoites stage (life cycle) in Pneumocystis as reported. Currently, 13 million chemical compounds are available for virtual screening in ZINC database. The biological information of four known drug molecules like TMP/SMX, Dapsone, Atovaquone and Pentamidine were collected from the PubChem compound database. Q-Site Finder online tool was used to determine the active site of DHFR in P. jiroveci. LogP values of chemical compounds were identified with the Atom-additive method. Since, existing drugs are synthetic chemicals that give more side effects in Pneumocystis affected patients. Polar surface area value of oxamide (86.18) was predicted to be in the ranges of existing drug values. Pentamidine was proved to be a more efficient ligand based on the dock score of -26.3398 still could not be considered as the natural compound oxamide also was highly comparable with the value of -20.3173. The binding affinity of the selected molecule was analyzed through Pose View and LigPlot.