ascorbic-acid and 5-methyltetrahydrofolate

To investigate the effect of simultaneous administration of [6S]-5-methyltetrahydrofolic acid ([6S]-5-CH(3)H(4)PteGlu) with L-ascorbic acid (L-AA) on serum folate concentrations in healthy male subjects.. A total of nine healthy male volunteers were recruited. Serum folate concentrations were measured before and up to 8 h after administration of each treatment (1) placebo, (2) 343 microg [6S]-5-CH(3)H(4)PteGlu), (3) 343 microg [6S]-5-CH(3)H(4)PteGlu) with 289.4 mg L-AA and (4) 343 microg [6S]-5-CH(3)H(4)PteGlu) with 973.8 mg L-AA (n=10 samples per treatment).. Serum folate concentrations significantly increased compared with baseline values, starting from 30 min after [6S]-5-CH(3)H(4)PteGlu administration and remained significantly higher than baseline values during the first 6 h for treatments 3 and 4, and during the first 4 h for treatment 2. Maximal serum folate responses were observed between 0.5 and 1.5 h after [6S]-5-CH(3)H(4)PteGlu consumption and significantly differed between treatments 2 and 4 (P<0.05). When [6S]-5-CH(3)H(4)PteGlu was concurrently administered with 289.4 or 973.8 mg L-AA, the total serum folate response, calculated as the area under the curve (AUC), was significantly improved (46.5+/-4.0 and 53.0+/-4.0 vs 34.3+/-3.8 h nmol/l, P<0.05). No significant difference in AUC was found between the 289.4 and the 973.8 mg L-AA treatments.. Administration of a physiological dose of [6S]-5-CH(3)H(4)PteGlu with L-AA significantly improved the measured serum folate response in folate saturated healthy men.

Topics: Administration, Oral; Adult; Antioxidants; Area Under Curve; Ascorbic Acid; Biological Availability; Biomarkers; Cross-Over Studies; Dose-Response Relationship, Drug; Folic Acid; Humans; Intestinal Absorption; Male; Tetrahydrofolates; Vitamin B Complex; Young Adult

Here we report on the comparative stability of free and microencapsulated L-5-methyltetrahydrofolate (L-5-MTHF) with free folic acid (FA) when exposed to thermal cooking conditions that are common to noodle making. Fortifying noodle flour with free L-5-MTHF produced the greatest loss of the vitamin when noodles were cooked. In contrast, the percentage recovery of microencapsulated L-5-MTHF in both fresh and cooked noodles was not significantly different to noodles that were similarly processed with fortified FA. The addition of sodium ascorbate along with L-5-MTHF enabled a sustained stability of the folate after boiling, and also after frying. The dispersal of microencapsulated folate in flour showed better homogeneity compared to similar practices used with free form L-5-MTHF and FA, respectively. We conclude that microencapsulating L-5-MTHF along with sodium ascorbate is effective to produce a stable folate in fortified noodles, a staple food for Asian populations that may require improved dietary folate intake.

Topics: Ascorbic Acid; Biological Availability; Cooking; Drug Compounding; Flour; Folic Acid; Food Analysis; Tetrahydrofolates; Water

The C1561T variant of the glutamate carboxypeptidase II (GCPII) gene is critical for natural methylfolylpolyglutamte (methylfolate) absorption, and has been associated with perturbations in folate metabolism and disease susceptibility. However, little is known on C1561T-GCPII as a risk factor for colorectal cancer. Therefore, this study examined whether C1561T-GCPII influences folate metabolism and adenomatous polyp occurrence.. 164 controls and 38 adenomatous polyp cases were analysed to determine blood folate and plasma homocysteine (Hcy) level, dietary intake of natural methylfolate, synthetic pteroylglutamic acid (PteGlu), vitamin C and C1561T-GCPII genotype.. In controls and cases, 7.3 and 18.4 percent of subjects respectively, were found to have the CT genotype, increasing the risk for adenomatous polyp occurrence 2.86 times (95% CI:1.37-8.0, p=0.035). Total dietary folate, methylfolate and PteGlu intake and the level of erythrocyte folate and plasma Hcy did not predict the occurrence of an adenomatous polyp. However, dietary natural vitamin C intake was associated with adenomatous polyp risk within C1561T-GCPII CT genotype subjects (p=0.037).. The findings suggest that C1561T-GCPII variation may be associated with risk for adenomatous polyp, and vitamin C may modify risk by interacting with the variant gene, its expression product and/or folate substrates.

Topics: Adenomatous Polyps; Adult; Aged; Aged, 80 and over; Ascorbic Acid; Case-Control Studies; Colorectal Neoplasms; Diet; Folic Acid; Genotype; Glutamate Carboxypeptidase II; Homocysteine; Humans; Middle Aged; Polymorphism, Single Nucleotide; Pteroylpolyglutamic Acids; Risk Factors; Tetrahydrofolates; Vitamins

Deficiency of 5-methyltetrahydrofolate (5-MTHF) in cerebrospinal fluid (CSF) is associated with a number of neurometabolic conditions including mitochondrial electron transport chain defects. Whilst failure of the active transport of 5-methyltetrahydrofolate (5-MTHF) into the CSF compartment has been proposed as a potential mechanism responsible for the 5-MTHF deficiency seen in mitochondrial disorders, it is becoming increasingly clear that other mechanisms are involved. Here, we have considered the role of oxidative stress as a contributing mechanism. Concerning, ascorbic acid (AA), we have established a CSF reference range (103-303μM) and demonstrated a significant positive correlation between 5-MTHF and AA. Furthermore, CSF itself was also shown to convey antioxidant properties towards 5-MTHF. However, this protection could be overcome by the introduction of a hydroxyl radical generating system. Using a neuronal model system, inhibition of mitochondrial complex I, by 58%, was associated with a 23% increase in superoxide generation and a significantly increased loss of 5-MTHF from the extracellular medium. Addition of AA (150μM) was able to prevent this increased 5-MTHF catabolism. We conclude that increased generation of reactive oxygen species and/or loss of CSF antioxidants are also factors to consider with regard to the development of a central 5-MTHF deficiency. Co-supplementation of AA together with appropriate folate replacement may be of therapeutic benefit.

Topics: Adolescent; Adult; Ascorbic Acid; Cell Line, Tumor; Child; Child, Preschool; Female; Folic Acid; Humans; Infant; Infant, Newborn; Male; Middle Aged; Mitochondria; Reactive Oxygen Species; Tetrahydrofolates; Young Adult

Fortification of foods with L-5-methyltetrahydrofolic acid (L-5-MTHF) is challenging due to low stability to environmental conditions that include exposure to pH, moisture, and temperature. The objective of the present study was to stabilize L-5-MTHF using microencapsulation technology. L-5-MTHF microcapsules constructed with different core-to-wall ratios of L-5-MTHF, both alone or in combination with sodium ascorbate, yielded high (>89%) recovery of L-5-MTHF. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) analysis confirmed successful encapsulation of L-5-MTHF with high core-to-wall ratios. Microencapsulation of L-5-MTHF alone with a high core-to-wall ratio significantly (p < 0.05) improved the stability of L-5-MTHF over the course of bread baking, performed both in pilot plant and in commercial baking conditions. Breads made with fortified flour containing sodium ascorbate coencapsulated with L-5-MTHF had recoveries of L-5-MTHF that were 97% and 77%, respectively, for pilot plant and bakery breads. Co-encapsulating L-5-MTHF with ascorbate also significantly (p < 0.05) improved stability during storage, as compared to breads that contained free L-5-MTHF.

Topics: Ascorbic Acid; Bread; Chromatography, High Pressure Liquid; Color; Spectrometry, Mass, Secondary Ion; Tetrahydrofolates

The thermal stability of L-5-methyltetrafolic acid (L-5-MTHF) was investigated in model/buffer systems and food systems. L-5-MTHF degradation followed first-order reaction kinetics with relatively greater (P < 0.01) stability at pH 4 compared to pH 6.8 in the buffer systems. This was confirmed using cyclic voltammetry. The stability (for example, k-values) of L-5-MTHF in an oxygen controlled environment improved (P < 0.001) proportionally when in the presence of increasing molar ratios of sodium ascorbate (NaAsc). The addition of NaAsc to L-5-MTHF after heat treatment was also effective at returning thermally oxidized L-5-MTHF back to its original form. A scheme was developed to explain the degradation and regeneration of L-5-MTHF. The importance of antioxidant protection of L-5-MTHF from thermal oxidation was extended using 2 distinct food systems; namely skim milk and soy milk, both with known antioxidant capacities. We conclude that the antioxidant activity of food components can enhance the stability of L-5-MTHF.

Topics: Antioxidants; Ascorbic Acid; Dietary Supplements; Food Handling; Hot Temperature; Kinetics; Oxidation-Reduction; Tetrahydrofolates

Tetrahydrobiopterin has been shown to efficiently abrogate ischemia reperfusion injury (IRI). However, it is unclear, whether its beneficial action relies on cofactor activity of one of the five known tetrahydrobiopterin-dependent reactions or on its antioxidative capacity. We therefore compared tetrahydrobiopterin with the pterin derivate tetrahydroneopterin (similar biochemical properties, but no nitric oxide synthase cofactor activity) and the antioxidants vitamin C and 5-methyltetrahydrofolate. Donor mice were pretreated with tetrahydrobiopterin, tetrahydroneopterin, vitamin C, or 5-methyltetrahydrofolate. Pancreatic grafts were subjected to 16-h cold ischemia time and implanted in syngeneic recipients. Untreated and nontransplanted animals served as controls. Following 2-h reperfusion, microcirculation was analyzed by intravital fluorescence microscopy. Graft damage was assessed by histology and nitrotyrosine immunostaining, and tetrahydrobiopterin levels were determined by HPLC. Recipient survival served as ultimate readout. Prolonged cold ischemia time resulted in microcirculatory breakdown. Only tetrahydrobiopterin pretreatment succeeded to preserve the capillary net, whereas all other compounds showed no beneficial effects. Along with increased intragraft tetrahydrobiopterin levels during recovery and implantation, only tetrahydrobiopterin pretreatment led to significant reduction of IRI-related parenchymal damage enabling recipient survival. These results show a striking superiority of tetrahydrobiopterin in preventing lethal IRI compared with related compounds and suggest nitric oxide synthases as treatment target.

Topics: Animals; Antioxidants; Ascorbic Acid; Biopterins; Cold Ischemia; Immunohistochemistry; Ischemia; Liver; Male; Mice; Mice, Inbred C57BL; Microcirculation; Microscopy, Confocal; Nitric Oxide; Organ Preservation; Pancreas; Pancreas Transplantation; Reperfusion Injury; Tetrahydrofolates; Time Factors

Bioactive compounds and their relationship with antioxidant activity were determined in three tomato cultivars (Ronaldo, Siena and Copo) during vine ripening. The lycopene, chlorophyll (total, a and b), total phenolic, flavonoid, vitamin C and folate contents, and the antioxidant activity, by the ferric reducing/antioxidant power assay and the beta-carotene lineolate system, were determined in the samples. Tomato ripening involved the breakdown of chlorophylls, accompanied by a continuous increase in the lycopene content. Total phenolics, flavonoids and vitamin C increased significantly during ripening, whereas the folate content fell markedly as tomatoes turned from green to red. The lycopene and flavonoid content was highest in the Copo cultivar, vitamin C and folate highest in Ronaldo, and total phenolics highest in Siena. The antioxidant activity, as measured with the ferric reducing/antioxidant power assay, increased significantly during ripening in all extracts, and showed a positive correlation with the total phenolic and flavonoid contents. However, when measured with the beta-carotene lineolate system, the antioxidant activity decreased significantly during ripening; perhaps due to the antioxidant activity of chlorophylls and the peroxidation activity of vitamin C.

Topics: Antioxidants; Ascorbic Acid; Carotenoids; Chlorophyll; Flavonoids; Fruit; Lycopene; Oxidation-Reduction; Phenols; Pigmentation; Plant Extracts; Solanum lycopersicum; Species Specificity; Tetrahydrofolates

The known functions of folate are to support one-carbon metabolism and to serve as photoreceptors for cryptochromes and photolyases. We demonstrate that 5-methyltetrahydrofolate (5-MTHF, the predominant folate in plasma) is also a potent, near diffusion limited, scavenger of singlet oxygen and quencher of excited photosensitizers. Both pathways result in decomposition of 5-MTHF, although ascorbate can protect against this loss. In the absence of photosensitizers, 5-MTHF is directly decomposed only very slowly by UVA or UVB. Although synthetic folic acid can promote DNA damage by UVA, submicromolar 5-MTHF inhibits photosensitization-induced strand breaks. These observations suggest a new role for reduced folate in protection from ultraviolet damage and have bearing on the hypothesis that folate photodegradation influenced the evolution of human skin color.

Topics: Ascorbic Acid; Chromatography, High Pressure Liquid; Depression, Chemical; DNA Breaks; DNA Damage; DNA, Superhelical; Folic Acid; Free Radical Scavengers; Oxidation-Reduction; Pentetic Acid; Photochemistry; Photosensitizing Agents; Pteridines; Rose Bengal; Singlet Oxygen; Sodium Azide; Superoxide Dismutase; Tetrahydrofolates; Ultraviolet Rays

The endothelial dysfunction induced by hyperhomocysteinemia can be reversed by 5-methyltetrahydrofolate (5-MTHF) via homocysteine (Hcy) lowering. An additive antioxidant action of 5-MTHF has been suggested to ameliorate endothelial dysfunction through increased nitric oxide production and superoxide radical scavenging, independent of Hcy lowering. The aim of the study was to assess whether 5-MTHF affects the redox state in hyperhomocysteinemia. We examined the effect of 3 months of oral 5-MTHF treatment (15 mg/day) on the redox pattern in 48 hyperhomocysteinemic subjects compared to 24 untreated hyperhomocysteinemic subjects. By analysis of variance with repeated measures in the 72 subjects, 5-MTHF markedly decreased plasma total Hcy (p-tHcy; P = 0.0001) and blood-total glutathione (GSH; b-tGSH; P = 0.002). By multivariate linear regression in the treated subjects, p-tHcy changes from baseline to 3 months (adjusted by baseline p-tHcy levels) correlated only with changes in reduced cysteinylglycine (P = 0.001). The effects of treatment on Hcy lowering and GSH metabolism were greater in medium than in moderate hyperhomocysteinemia. In conclusion, high-dose 5-MTHF treatment for 3 months ensures marked Hcy lowering to normal values even in subjects with high Hcy levels, and should be the treatment of choice in medium hyperhomocysteinemia. Furthermore, 5-MTHF shows a favorable interaction with GSH metabolism.

Topics: Adult; Antioxidants; Ascorbic Acid; Chromatography, High Pressure Liquid; DNA; Female; Folic Acid; Homocysteine; Humans; Hyperhomocysteinemia; Lipid Peroxidation; Male; Malondialdehyde; Middle Aged; Oxidation-Reduction; Sulfhydryl Compounds; Tetrahydrofolates; Vitamin B 12; Vitamin E

A comparative study on the pressure and temperature stability of 5-methyltetrahydrofolic acid (5-CH(3)-H(4)folate) was performed in model/buffer systems and food products (i.e., orange juice, kiwi puree, carrot juice, and asparagus). Effects of pH and ascorbic acid (0.5 mg/g) on 5-CH(3)-H(4)folate stability in buffer systems were studied on a kinetic basis at different temperatures (from 65 to 160 degrees C) and different pressure/temperature combinations (from 100 to 700 MPa/from 20 to 65 degrees C). These studies showed that (i) the degradation of 5-CH(3)-H(4)folate in all model systems could be described by first-order reaction kinetics, (ii) the thermostability of 5-CH(3)-H(4)folate was enhanced by increasing pH up to 7, (iii) 5-CH(3)-H(4)folate was relatively pressure stable at temperatures lower than 40 degrees C, and (iv) ascorbic acid enhanced both the thermo- and barostabilities of 5-CH(3)-H(4)folate. In food products, temperature and pressure stabilities of 5-CH(3)-H(4)folate were studied at different temperatures (70-120 degrees C) and different pressure/temperature combinations (from 50 to 200 MPa/25 degrees C and 500 MPa/60 degrees C). 5-CH(3)-H(4)folate in orange juice and kiwi puree was relatively temperature (up to 120 degrees C) and pressure (up to 500 MPa/60 degrees C) stable in contrast to carrot juice and asparagus. Addition of ascorbic acid (0.5 mg/g) in carrot juice resulted in a remarkable protective effect on pressure (500 MPa/60 degrees C/40 min) and temperature degradation (120 degrees C/40 min) of 5-CH(3)-H(4)folate.

Topics: Ascorbic Acid; Beverages; Drug Stability; Food; Fruit; Hydrogen-Ion Concentration; Pressure; Temperature; Tetrahydrofolates; Vegetables

We sought to investigate the effects of short- and long-term vitamin C therapy on endothelial dysfunction in patients with homocystinuria.. Untreated homocystinuria due to cystathionine beta-synthase deficiency is associated with premature atherothrombotic disease; 25% of untreated patients suffer a vascular event by the age of 16 years and 50% by 29 years. Treatment directed at reducing homocysteine accumulation significantly reduces this risk. However, despite 'optimal' treatment and compliance, hyperhomocysteinaemia usually persists and individuals exhibit endothelial dysfunction indicative of an adverse cardiovascular prognosis. Additional intervention is therefore required to further reduce cardiovascular risk.. We investigated the endothelial effects of acute (2 g single dose) and chronic (1 g/day for 6 months) administration of oral vitamin C in 5 patients with homocystinuria (mean age 26 years, 1 male) and 5 age- and sex-matched controls. Brachial artery endothelium-dependent flow-mediated dilatation (FMD) and endothelium-independent responses to nitroglycerin (NTG) were measured using high-resolution ultrasonic vessel wall-tracking.. Baseline: Plasma total homocysteine was 100.8 +/- 61.6 and 9.2 +/- 1.9 micromol/L in the patient and control groups, respectively (p < 0.001). FMD responses were impaired in the patient group (20 +/- 40 microm) compared with the controls (116 +/- 30 microm) (p < 0.001). Vitamin C administration: FMD responses in the patient group improved both acutely, 160 +/- 65 microm at 4 h (p < 0.001), and chronically, 170 +/- 70 microm at 2 weeks (p < 0.001) and 170 +/- 40 microm at 6 months (p < 0.001). FMD responses in the control group were unaltered (p = 0.526). Within both groups, neither the vascular response to NTG nor plasma homocysteine was altered (p > 0.4).. Vitamin C ameliorates endothelial dysfunction in patients with homocystinuria, independent of changes in homocysteine concentration and should therefore be considered as an additional adjunct to therapy to reduce the potential long-term risk of atherothrombotic disease.

Topics: Adult; Ascorbic Acid; Blood Flow Velocity; Blood Pressure; Brachial Artery; Endothelium, Vascular; Female; Heart Rate; Homocystine; Homocystinuria; Humans; Male; Methionine; Nitric Oxide; Nitroglycerin; Tetrahydrofolates; Vasodilation

Binding of folate (pteroylglutamate) and 5-methyltetrahydrofolate, the major endogenous form of folate, to folate binding protein purified from cow's milk was studied at 7 degrees C to avoid degradation of 5-methyltetrahydrofolate. Both folates dissociate rapidly from the protein at pH 3.5, but extremely slowly at pH 7.4, most likely due to drastic changes in protein conformation occurring after folate binding. Dissociation of 5-methyltetrahydrofolate showed no increase at 37 degrees C suggesting that protein-bound-5-methyltetrahydrofolate is protected against degradation. Binding displayed two characteristics, positive cooperativity and a binding affinity that increased with decreasing concentrations of the protein. The binding affinity of folate was somewhat greater than that of 5-methyl tetrahydrofolate, in particular at pH 5.0. Ligand-bound protein exhibited concentration-dependent polymerization (8-mers formed at 13 microM) at pH 7.4. At pH 5.0, only folate-bound forms showed noticeable polymerization. The fact that folate at pH 5.0 surpasses 5-methyltetrahydrofolate both with regard to binding affinity and ability to induce polymerization suggests that ligand binding is associated with conformational changes of the protein which favor polymerization.

Topics: Animals; Ascorbic Acid; Carbon Radioisotopes; Carrier Proteins; Folate Receptors, GPI-Anchored; Folic Acid; Hydrogen-Ion Concentration; Milk; Polymers; Radioligand Assay; Receptors, Cell Surface; Tetrahydrofolates

Erythrocyte (RBC) folates occur mainly as 5-methyltetrahydrofolate polyglutamates. Determination of RBC folate concentration requires an initial deconjugation of these polyglutamates. In this study, existing HPLC methods were adapted to investigate the rate and extent of this deconjugation process. The action of endogenous plasma pteroyl-polyglutamate hydrolase activity was strongly affected by the conditions of sample preparation, with pH of the incubation mixture more critical to effective deconjugation than incubation time. Dilution of whole blood with 10 g/L ascorbic acid yielded fast hydrolysis of long-chain polyglutamates, and total conversion to 5-methyltetrahydrofolate monoglutamate occurred after 90 min of incubation at 37 degrees C. In contrast, dilution of whole blood with 10 g/L sodium ascorbate, with up to 90 min of incubation at 37 degrees C, yielded a mixture of polyglutamates of 5-methyltetrahydrofolate (glun = 1-8). As documented by direct HPLC analysis and in concurrent assays with Lactobacillus casei, acidification provided by ascorbic acid can have dramatic effects on the measurement of RBC folates.

Topics: Ascorbic Acid; Chromatography, High Pressure Liquid; Erythrocytes; Folic Acid; gamma-Glutamyl Hydrolase; Humans; Hydrogen-Ion Concentration; Hydrolysis; Kinetics; Pteroylpolyglutamic Acids; Tetrahydrofolates

We investigated the oxidative degradation pathway of 5CH3-H4PteGlu, the main extracellular folate and the predominant form of the vitamin found in food and blood. 5CH3-H4PteGlu is oxidized to 5CH3-5,6-H2PteGlu which subsequently undergoes C9-N10 bond cleavage yielding a pteridine residue and P-ABG, the latter step resulting in irreversible loss of vitamin activity. Under moderately acid conditions typical of the postprandial gut (pH 3.5) 5CH3-H4PteGlu is fairly stable (t1/2 = 273.6 min), while 5CH3-5,6-H2PteGlu is rapidly degraded (t1/2 = 16.9 min). In a neutral environment (pH 6.4) stability is reversed; 5CH3-H4PteGlu t1/2 = 12.0 mins, 5CH3-5,6-H2PteGlu t1/2 = 1504.6 min. Ascorbic acid was efficacious in the facile salvage of 5CH3-H4PteGlu from 5CH3-5,6-H2PteGlu which occurred rapidly and with significant efficiency (100% conversion) under acid (pH 3.5) conditions, t1/2 = 1.3 min (1 mmol/liter ascorbate), but was less efficient under neutral (pH 6.4) conditions t1/2 = 273.6 min (36% conversion). The presence of zinc and iron broadly maintains the pattern of effect, but increases all reaction rates. PteGlu was stable under all conditions studied. These results obtained in an artificial environment were supported by findings in human gastric juice: at a gastric pH of 1.47 with low endogenous ascorbate (7.0 mumol/liter), 5CH3-5,6-H2PteGlu and 5CH3-H4PteGlu both degrade instantly via C9-N10 bond cleavage to yield an equimolar amount of P-ABG. If the same gastric juice is spiked at 58.0 mumol/liter ascorbate (moderate endogenous concentration), 5CH3-H4PteGlu is stable (t1/2 = 334.7 min), while 5CH3-5,6-H2PteGlu is instantly salvaged to 5CH3-H4PteGlu with 43.3% efficiency, and the remaining 5CH3-5,6-H2PteGlu is degraded to P-ABG. In gastric juice with an elevated pH of 7.0 and no endogenous ascorbate, 5CH3-5,6-H2PteGlu and 5CH3-H4PteGlu are both stable, with no C9-N10 bond cleavage. This, for 5CH3-H4PteGlu, is in apparent contrast to findings at pH 6.4 in an artificial environment. The same gastric juice spiked to 50 mumol/liter ascorbate did not result in 5CH3-H4PteGlu salvage from 5CH3-5,6-H2PteGlu.(ABSTRACT TRUNCATED AT 400 WORDS)

Topics: Ascorbic Acid; Biological Availability; Chemical Phenomena; Chemistry, Physical; Diet; Folic Acid; Gastric Juice; Humans; Hydrogen-Ion Concentration; In Vitro Techniques; Kinetics; Oxidation-Reduction; Tetrahydrofolates

1. The response of Lactobacillus casei was measured for a number of the monoglutamyl forms of folate derivatives. 2. At the concentrations of folate commonly used in the assay of folate vitamin in foods the response of L. casei to folic acid, (pteroylglutamic acid) and 5-formyl-tetrahydrofolic acid was similar, but 5-methyl-tetrahydrofolic acid gave as little as half the response of folic acid. 3. The response was modified by altering pH but not by concentration of ascorbate. 4. These results have implications for the assays of foods for folate where mixtures of folate derivatives are present. 5. A modified procedure is suggested in which the monoglutamates give similar responses.

Topics: Ascorbic Acid; Biological Assay; Culture Media; Folic Acid; Food Analysis; Formyltetrahydrofolates; Hydrogen-Ion Concentration; Lacticaseibacillus casei; Structure-Activity Relationship; Tetrahydrofolates

The rescue of lymphocytes from methotrexate (MTX) growth inhibition by 5-methyltetrahydrofolate (5-methyl-THF) and 5-formyltetrahydrofolate (5-formyl-THF) has been studied. Rescue by 5-methyl-THF is selective for cells with high levels of homocysteine:5-methyl-THF methyl-transferase (methyltransferase). At MTX concentrations which inhibited growth greater than or equal to 85% in both leukemic T-lymphocytes (CCRF-CEM) and Epstein-Barr-transformed B-lymphocytes (LAZ-007), 5 micro M 5-formyl-THF rescued more effectively than did 5-methyl-THF, in either the presence or absence of the methyltransferase inhibitor, nitrous oxide. At less inhibitory MTX concentrations, both reduced folates rescued equally, except when methyltransferase was inhibited by nitrous oxide in which case 5-formyl-THF was clearly superior. In the absence of nitrous oxide, both cell lines contained approximately equal amounts of methyltransferase. Some apparent differences in the rescue of these cell lines with 5-methyl-THF were attributable to their different sensitivity to MTX. When metabolism of reduced folates was severely impaired by MTX and nitrous oxide, lymphocytes were rescued with 5-[methyl-14C]methyl-THF, and the uptake of 14C into DNA was measured. In corporation was very low, indicating that cellular oxidation of 5-methyl-THF to 5,10-methylene-tetrahydrofolate is minimal even under forcing conditions. MTX selectively in vivo will be influenced by the level of methyltransferase in tumor and normal tissues.

Topics: 5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase; Ascorbic Acid; Cell Line; DNA; Humans; Leucovorin; Lymphocytes; Methotrexate; Nitrous Oxide; Oxidation-Reduction; Tetrahydrofolates

Topics: Animals; Ascorbic Acid; Biopterins; Child, Preschool; Humans; Hydroxocobalamin; In Vitro Techniques; Infant; Leucovorin; Male; NADH, NADPH Oxidoreductases; Phenylketonurias; Rats; Tetrahydrofolates