Search

Highlights • Exogenous phenylalanine application confers chilling tolerance in tomato fruits. • Conferring chilling tolerance is accompanying by maintaining membrane integrity. • Employing higher MTHFD enzyme activity is crucial for ensuring sufficient supply of NADPH. • Higher ROS scavenging system activity may arise from ensuring sufficient supply of NADPH. • Ensuring sufficient supply of NADPH giving rise to higher phenols and flavonoids accumulation and DPPH scavenging capacity. Abstract In this experiment, the mechanisms displayed by exogenous phenylalanine application to confer chilling tolerance in tomato fruits (Lycopersicon esculentum cv. Izmir) during storage at 4 °C for 28 days were studied. Phenylalanine application at 5 mM significantly confers chilling injury tolerance in tomato fruits associated with membrane integrity maintenance reflected by a lower degree of electrolytes leakage and malondialdehyde (MDA) accumulation. Membrane integrity maintenance in tomato fruits in response to exogenous phenylalanine application at 5 mM may arise from lower H 2 O 2 accumulation due to higher activity of reactive oxygen species (ROS) scavenging enzymes superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR), concurrent with higher endogenous lycopene and proline accumulation. In addition, higher phenols and flavonoids accumulation arising from higher phenylalanine ammonia-lyase (PAL) enzyme activity is concurrent with higher ascorbic acid (AA) accumulation giving rise to higher DPPH scavenging capacity, which may be vital for the maintenance of membrane integrity in tomato fruits treated with exogenous phenylalanine. Ensuring supply of reducing power NADPH by triggering of folates pathway demonstrated by higher methylenetetrahydrofolate dehydrogenase (MTHFD) enzyme activity may be responsible for activation of ROS scavenging systems in tomato fruits in response to exogenous phenylalanine application, contributing to membrane integrity maintenance. Accordingly, exogenous phenylalanine application may serve as a safe promising procedure to confer chilling tolerance in tomato fruits during cold storage. [ABSTRACT FROM AUTHOR]

Background Multi-target approaches are necessary to properly analyze or modify the function of a biochemical pathway or a protein family. An example of such a problem is the repurposing of the known human anti-cancer drugs, antifolates, as selective anti-parasitic agents. This requires considering a set of experimentally validated protein targets in the folate pathway of major pathogenic trypanosomatid parasites and humans: ( i ) the primary parasite on-targets: pteridine reductase 1 (PTR1) (absent in humans) and bifunctional dihydrofolate reductase-thymidylate synthase (DHFR–TS), ( ii ) the primary off-targets: human DHFR and TS, and ( iii ) the secondary on-target: human folate receptor β , a folate/antifolate transporter. Methods We computationally compared the structural, dynamic and physico-chemical properties of the targets. We based our analysis on available inhibitory activity and crystallographic data, including a crystal structure of the bifunctional T. cruzi DHFR–TS with tetrahydrofolate bound determined in this work. Due to the low sequence and structural similarity of the targets analyzed, we employed a mapping of binding pockets based on the known common ligands, folate and methotrexate. Results Our analysis provides a set of practical strategies for the design of selective trypanosomatid folate pathway inhibitors, which are supported by enzyme inhibition measurements and crystallographic structures. Conclusions The ligand-based comparative computational mapping of protein binding pockets provides a basis for repurposing of anti-folates and the design of new anti-trypanosmatid agents. General significance Apart from the target-based discovery of selective compounds, our approach may be also applied for protein engineering or analyzing evolutionary relationships in protein families. [ABSTRACT FROM AUTHOR]

In the current study, an artificial neural network (ANN)-based breast cancer prediction model was developed from the data of folate and xenobiotic pathway genetic polymorphisms along with the nutritional and demographic variables to investigate how micronutrients modulate susceptibility to breast cancer. The developed ANN model explained 94.2% variability in breast cancer prediction. Fixed effect models of folate (400 μg/day) and B 12 (6 μg/day) showed 33.3% and 11.3% risk reduction, respectively. Multifactor dimensionality reduction analysis showed the following interactions in responders to folate: RFC1 G80A × MTHFR C677T (primary), COMT H108L × CYP1A1 m2 (secondary), MTR A2756G (tertiary). The interactions among responders to B 12 were RFC1G80A × cSHMT C1420T and CYP1A1 m2 × CYP1A1 m4. ANN simulations revealed that increased folate might restore ER and PR expression and reduce the promoter CpG island methylation of extra cellular superoxide dismutase and BRCA1. Dietary intake of folate appears to confer protection against breast cancer through its modulating effects on ER and PR expression and methylation of EC-SOD and BRCA1. [ABSTRACT FROM AUTHOR]

Genetic factors may play an important role in susceptibility to childhood acute lymphoblastic leukemia (ALL). The aim of our study was to evaluate the associations of genetic polymorphisms in folate pathway and DNA repair genes with susceptibility to ALL. In total, 121 children with ALL and 184 unrelated healthy controls of Slovenian origin were genotyped for 14 polymorphisms in seven genes of folate pathway, base excision repair and homologous recombination repair ( TYMS , MTHFR , OGG1 , XRCC1 , NBN , RAD51 , and XRCC3 ). In addition, the exon 6 of NBN was screened for the presence of mutations using denaturing high performance liquid chromatography. Twelve polymorphisms were in Hardy–Weinberg equilibrium in controls and their genotype frequencies were in agreement with those reported in other Caucasian populations. Among the investigated polymorphisms and mutations, NBN Glu185Gln significantly decreased susceptibility to B-cell ALL ( p = 0.037), while TYMS 3R allele decreased susceptibility to T-cell ALL ( p = 0.011). Moreover, significantly decreased susceptibility to ALL was observed for MTHFR TA ( p = 0.030) and RAD51 GTT haplotypes ( p = 0.016). Susceptibility to ALL increased with the increasing number of risk alleles ( p trend = 0.007). We also observed significant influence of hOGG – RAD51 and NBN – RAD51 interactions on susceptibility to ALL. Our results suggest that combination of several polymorphisms in DNA repair and folate pathways may significantly affect susceptibility to childhood ALL. [ABSTRACT FROM AUTHOR]

Abstract: Objectives: This study aims to determine the possible association between folate pathway gene polymorphisms and idiopathic pulmonary fibrosis. This represents the first study carried out on folate pathway gene polymorphisms as possible risk factors in this kind of pathology. The premise is that several polymorphisms mapping on genes responsible for folate uptake are associated with the risk of numerous diseases occurring between pregnancy and old age, and that too little is currently known about idiopathic pulmonary fibrosis. Design and methods: We genotyped 9 single nucleotide polymorphisms and 1 polymorphic insertion in 7 essential genes belonging to the folate pathway in 32 Italian idiopathic pulmonary fibrosis patients and 81 control subjects. This was done by PCR and restriction analysis. Results: Allelic and genotypic association tests indicated that for all the analysed polymorphisms there were no significant differences between patients and controls. Nevertheless, the haplotype association analysis revealed a significant association between idiopathic pulmonary fibrosis and transcobalamin II gene polymorphisms: specifically the haplotype 776G (rs1801198)–c.1026-394G (rs7286680)–444C (rs10418) (OR=2.84; 95% C.I. 1.36–5.93, P value=0.004). Conclusions: This small-scale preliminary study would suggest the importance of further research focusing on the role of folate in the onset of idiopathic pulmonary fibrosis. [Copyright &y& Elsevier]

Abstract: Folate, a vitamin of the B group involved in one-carbon group metabolism, plays an important role in DNA synthesis and methylation. Several polymorphisms in the genes involved in folate uptake and biotransformations have been shown to be associated to the risk of cancer and to anticancer drug response. We studied common polymorphisms in MTHFR (N5,10-methylene-tetrahydrofolate reductase), MTHFD1 (N5,10-methylene-tetrahydrofolate dehydrogenase), MTR (methionine synthetase) and SLC19A1 (reduced folate carrier) in the panel of 60 human tumour cell lines established by the NCI for anticancer drug screening and we tentatively associated these polymorphisms with gene expression and drug cytotoxicity as extracted from the public database of the Developmental Therapeutic Programme. We observed a consistent and highly significant association between the presence of the variant C allele of the A>C1298 polymorphism of MTHFR and the sensitivity to many anticancer drugs belonging to the classes of antifolates, antimetabolites, alkylating agents and, to a lesser extent, topoisomerase inhibitors. In contrast, the T variant allele of the C>T677 variation of MTHFR was rather associated to lower sensitivity of the cell lines towards anticancer drugs (alkylating agents, antifolates and antimetabolites) but with much lower effects than the A>C1298 variation. The polymorphisms of the other genes studied were not associated with differences in anticancer drug sensitivity, but the expression of the SLC19A1 gene was significantly correlated with the sensitivity to several drugs (antifolates, thiopurines, nitrosoureas, and DACH-platinum drugs). We concluded that the NCI-60 panel may constitute a good starting point for implementing clinical studies aimed at discovering and validating predictive genetic markers of drug efficacy and/or toxicity. [Copyright &y& Elsevier]

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

With the background of association of oxidative stress and Hepatitis E virus (HEV) infection in pregnancy complications the present novel study aimed to evaluate the significance of changes in maternal homocysteine levels and the related mechanism(s) in the pathophysiology of HEV related pregnancy complications and negative outcomes. Term delivery (TD, N = 194) and HEV-IgM positive pregnancy cases [ N = 109] were enrolled. Serum and placental homocysteine levels were evaluated by ELISA and immunofluorescence and in turn correlated with serum Vitamin B12 levels. Distribution of variant MTHFR C➔T and TYMS1494del6bp genotyping were studied by PCR-RFLP. Differential folate receptor alpha (FR-α) expression in placenta was evaluated by real-time PCR and immunofluorescence respectively. The HEV viral load was significantly higher in both FHF and AVH cases. Higher serum homocysteine levels was associated with preterm delivery (PTD) and fetal death in HEV infected cases and was significantly inversely correlated with serum VitaminB12 levels in HEV cases. Placental homocysteine expression was upregulated in HEV cases, and in cases with negative pregnancy outcome. A Homocysteine level was associated with MTHFR C677T status. Genetic alterations in folate pathway was associated with increased risk of PTD in HEV infected pregnancy cases, disease severity, and negative pregnancy outcome in AVH and FHF groups. FR-α expression was downregulated in placental tissues of HEV infected pregnancy.Placental stress caused by HEV inflicted increased homocysteine due to alterations in maternal vitamin B12 levels and folate pathway components is detrimental mechanism in PTD and negative pregnancy outcome in HEV infected pregnancy cases and holds prognostic and therapeutic significance. • This is a first kind of novel study concerning oxidative stress in deciding the fate of HEV infected pregnancy cases. • High viral (HEV) load, Increased Homocysteine levels and decreased vitamin B12 levels are associated with pregnancycomplications. • MTHFR C677T and TYMS 1494del6bp polymorphism is associated with the preterm delivery and fetal death in HEV infected cases. • MTHFR C677T polymorphism associated with the increased homocysteine levels may have a prognostic significance. • Deregulation of folate receptor alpha (FR-α) expression is associated with preterm delivery and fetal death. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus (Sa) is a serious concern due to increasing resistance to antibiotics. The bacterial dihydrofolate reductase enzyme is effectively inhibited by trimethoprim, a compound with antibacterial activity. Previously, we reported a trimethoprim derivative containing an acryloyl linker and a dihydophthalazine moiety demonstrating increased potency against S. aureus. We have expanded this series and assessed in vitro enzyme inhibition (K i) and whole cell growth inhibition properties (MIC). Modifications were focused at a chiral carbon within the phthalazine heterocycle, as well as simultaneous modification at positions on the dihydrophthalazine. MIC values increased from 0.0626–0.5 μg/mL into the 0.5–1 μg/mL range when the edge positions were modified with either methyl or methoxy groups. Changes at the chiral carbon affected K i measurements but with little impact on MIC values. Our structural data revealed accommodation of predominantly the S -enantiomer of the inhibitors within the folate-binding pocket. Longer modifications at the chiral carbon, such as p -methylbenzyl, protrude from the pocket into solvent and result in poorer K i values, as do modifications with greater torsional freedom, such as 1-ethylpropyl. The most efficacious K i was 0.7 ± 0.3 nM, obtained with a cyclopropyl derivative containing dimethoxy modifications at the dihydrophthalazine edge. The co-crystal structure revealed an alternative placement of the phthalazine moiety into a shallow surface at the edge of the site that can accommodate either enantiomer of the inhibitor. The current design, therefore, highlights how to engineer specific placement of the inhibitor within this alternative pocket, which in turn maximizes the enzyme inhibitory properties of racemic mixtures. Image 1 • Trimethoprim derivatives offer additional chemical diversity within a proven scaffold. • Altering chemical moieties at the binding site: solvent interface modulates affinity. • Altering chemical moieties at the dihydrophthalazine edge alters cell inhibition. • Combining moieties improves inhibition by targeting a unique binding site surface. • Derivatives targeting this site do not prefer a specific enantiomer. [ABSTRACT FROM AUTHOR]