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Dehydroascorbic acid and ascorbic acid transport systems in the guinea pig ileum.

The transport properties of dehydroascorbic acid and ascorbic acid in membrane vesicle preparations of guinea pig ileum were evaluated. Na-dependent transport of ascorbic acid in the brush-border membrane was confirmed, and an Na-independent mechanism was found in the basolateral membrane. The electrically neutral oxidized form of vitamin C is transported by an Na-independent mechanism at both cell surfaces. Transport of each substrate is saturable and exhibits cis-inhibition and trans-stimulation in the presence of structural analogues. Additional studies on ascorbate metabolism will be necessary to support a comprehensive model of intestinal handling of vitamin C.

Animals↗

Proposed role for a combination of citric acid and ascorbic acid in the production of dietary iron overload: a fundamental cause of disease.

This paper presents a review of the significant body of literature liking dietary iron overload, not only to heart disease, but also to cancer, diabetes, osteoporosis, arthritis, and possibly other disorders. Following an analysis of our understanding of the mechanistic role iron plays in oxidative damage, an interpretation of the fact that plasma concentrations of several antioxidants are decreased in the presence of disease is offered. Evaluation of (1) age-related dietary trends over time and (2) factors involved in iron absorption leads to the hypothesis that the combination of citric acid and ascorbic acid (a synergistic pair of strong enhancers) is instrumental in causing a deleterious increase in iron load in aging populations. Iron overload may be the most important common etiologic factor in the development of the diseases mentioned; therefore, the synergistic combination of citric and ascorbic acids may play a major role in our worsening disease statistics. Evidence to support this hypothesis and possible experiments to test it are included. This combination needs further study, particularly because the iron overload produced may be correctable.

Aging↗

2-O-(beta-D-Glucopyranosyl)ascorbic acid, a novel ascorbic acid analogue isolated from Lycium fruit.

A novel stable precursor of ascorbic acid (vitamin C), 2-O-(beta-D-glucopyranosyl)ascorbic acid, was isolated from both the ripe fresh fruit and dried fruit of Lycium barbarum L., a plant of the Solanaceae family. The chemical structure was inferred by instrumental analyses and confirmed by chemical synthesis. The dried fruit of Lycium barbarum L. contained ca. 0.5% of it, which is comparable to the ascorbic acid content of fresh lemons. It increased the blood ascorbic acid by oral administration to rats, and it was also detected in blood from the portal vein.

Animals↗

Enhancing effect of 2-O-alpha-D-glucopyranosyl-L-ascorbic acid, a stable ascorbic acid derivative, on collagen synthesis.

The objective of the present study was to compare 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G) with ascorbic acid (AA) and ascorbic acid 2-phosphate (AA-2P) concerning the promotion of collagen production in human skin fibroblasts. Though AA-2G was still observed to be promoting collagen synthesis at the same level on the 8th day of the culture, collagen synthesis was seen to decrease on the fifth day of culturing with AA and AA-2P. This sustained collagen synthesis-promoting action is considered to be a major feature of the novel vitamin C derivative, AA-2G by conducting an experiment in which an alpha-glucosidase inhibitor was present, it was shown that AA-2G exerts its collagen synthesis-promoting action after being decomposed to AA by alpha-glucosidase. Further, we observed that for AA-2G, even on the 8th day of the culture, the amount of AA in the fibroblasts was virtually unchanged from the beginning of the experiment, whereas, in the case of adding AA and AA-2P, virtually no AA was detectable in the culture medium on the fifth day. These findings suggests that AA-2G is decomposed to AA by alpha-glucosidase in the cells. This AA promotes collagen synthesis, which is prolonged through AA-2G's sustained decomposition.

Ascorbic Acid↗

Protection and recycling of alpha-tocopherol in human erythrocytes by intracellular ascorbic acid.

Ascorbic acid can recycle alpha-tocopherol from the tocopheroxyl free radical in lipid bilayers and in micelles, but such recycling has not been demonstrated to occur across cell membranes. In this work the ability of intracellular ascorbate to protect and to recycle alpha-tocopherol in intact human erythrocytes and erythrocyte ghosts was investigated. In erythrocytes that were 80% depleted of intracellular ascorbate by treatment with the nitroxide Tempol, both 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) and ferricyanide oxidized alpha-tocopherol to a greater extent than in cells not depleted of ascorbate. In contrast, in erythrocytes in which the intracellular ascorbate concentration had been increased by loading with dehydroascorbate, loss of alpha-tocopherol was less with both oxidants than in control cells. Protection against AAPH-induced oxidation of alpha-tocopherol was not prevented by extracellular ascorbate oxidase, indicating that the protection was due to intracellular and not to extracellular ascorbate. Incubation of erythrocytes with lecithin liposomes also generated an oxidant stress, which caused lipid peroxidation in the liposomes and depleted erythrocyte alpha-tocopherol, leading to hemolysis. Ascorbate loading of the erythrocytes delayed liposome oxidation and decreased loss of alpha-tocopherol from both cells and from alpha-tocopherol-loaded liposomes. When erythrocyte ghosts were resealed to contain ascorbate and challenged with free radicals generated by AAPH outside the ghosts, intravesicular ascorbate was totally depleted over 1 h of incubation, whereas alpha-tocopherol decreased only after ascorbate was substantially oxidized. These results suggest that ascorbate within the erythrocyte protects alpha-tocopherol in the cell membrane by a direct recycling mechanism.

Amidines↗

Evolution and the need for ascorbic acid.

Ascorbic acid differs from other vitamins in that an exogenous source is required by only a few animal species. It is pointed out that this fact indicates that the amount contained in a diet of raw natural plant food is less than the optimum intake, corresponding to the best health. This argument leads to the conclusion that the optimum daily intake is about 2.3 g or more, for an adult with energy requirement 2500 kcal day(-1).

Adenosine Triphosphate↗

Effects of humic acids, para-aminobenzoic acid and ascorbic acid on the N-nitrosation of the carbamate insecticide propoxur and on the mutagenicity of nitrosopropoxur.

Nitrosation of the carbamate insecticide propoxur at pH 3 and 37 degrees C was determined colorimetrically and found to be time- and sodium nitrite concentration-dependent. Nitrosated propoxur was mutagenic when exposed to the seeds of the higher plant Arabidopsis thaliana but the formation of nitrosopropoxur, the presumed mutagen, was inhibited by humic acids, para-aminobenzoic acid and ascorbic acid. These agents also reduced the mutagenicity of preformed nitrosopropoxur.

4-Aminobenzoic Acid↗

Dental caries and mutans streptococci in relation to plasma ascorbic acid.

Ascorbic acid (AA) affects in vitro growth of bacteria and may also act in vivo to decrease caries activity. The aim of this study was to evaluate the possible association of AA level in plasma with number of caries lesions, relative numbers of some species of oral cariogenic flora, and rate of salivary secretion. The caries status and some bacteriologic variables of dentulous adult subjects with a low level of AA in the plasma (< or = 25 mumol/l; n = 75) were compared with those of controls (plasma level > or = 50 mumol/l; n = 75) matched for age, sex, and number of teeth. For each subject, site-specific recordings of the presence or absence of plaque, dental caries, fillings, and erosions were recorded clinically by the same dentist in a double-blind system. The amounts of visible plaque and numbers of decayed tooth surfaces were significantly higher in the low AA group than in the controls. No between-group differences were found in the number of fillings and the amount of oral bacterial growth. The frequencies of consumption of vegetables, berries, and other fruit were significantly lower in the low AA group than in the controls.

Adult↗

Quantification of L-ascorbic acid and total ascorbic acid in fruits and spinach by capillary zone electrophoresis.

A standard curve for the quantification of L-ascorbic acid (L-AA) by capillary zone electrophoresis (CZE) was established, and the quantification of ascorbic acid and total ascorbic acid in fruits (lemon, Sunkist, and pineapple) and spinach were performed using D-isoascorbic acid (D-IAA) as an internal standard. The minimum detection limits (MDLs) for L-AA and D-IAA were determined to be 1 and 2 microg/mL, respectively, at 265 nm. Dehydroascorbic acid (DHAA) in fruits and spinach was quantified in the presence of DL-homocysteine. The recoveries for L-AA in these juices were between 95 and 105%.

Ascorbic Acid↗

Uptake of dehydroascorbic acid and ascorbic acid to isolated nerve terminals and secretory granules from ox neurohypophyses.

When uptake of L-[14C]ascorbic acid ([14C]AA) to various organs in guinea-pigs was studied after intracardiac injection, the adenohypophysis, pars intermedia, and the neurohypophysis had an uptake per milligramme protein which was about half of the uptake to the adrenals. Adrenal uptake was 20 +/- 2.8 pmol mg-1 protein microCi-1 injected. The uptake to the different parts of the hypophysis was considerably higher than the uptake to pancreas, liver, kidney, spleen and other organs. When isolated nerve endings (neurosecretosomes) from ox neurohypophyses were incubated with a medium containing labelled dehydroascorbic acid ([14C]DHA), the uptake was much slower than when the medium contained labelled ascorbic acid. The uptake of [14C]DHA showed a linear dependence on concentration, and was not influenced by addition of Mg2+ and ATP. Addition of Mg2+ + ATP, omission of Ca2+ and Mg2+ or exchange of Na+ in the medium with K+ had no effect on the uptake of ascorbic acid. When isolated secretory granules from ox neurohypophyses were incubated with a medium containing [14C]DHA, uptake was considerably faster than the uptake when they were incubated in a medium containing [14C]AA. The uptake of dehydroascorbic acid was linear with the concentration in the medium and was not changed by addition of Mg2+ ATP. Addition of 10 mM NH4Cl or exchange of 120 mM K+ in the incubation medium with Na+ did not change the uptake of dehydroascorbic acid. The contents of copper, zinc, iron and cobalt were determined in isolated nerve endings (A) and membranes (B) as well as in lysate (C) from isolated neurosecretory granules. The results (in nmol mg-1 protein) were for Cu: (A): 0.25 +/- 0.01 (SEM), (B): 0.67 +/- 0.16, (C): 0.22 +/- 0.06; for Zn: (A): 0.53 +/- 0.13, (B): 6.97 +/- 0.75, (C): 1.8 +/- 0.53; and for Fe: (A): 15.6 +/- 1.9, (B): 6.92 +/- 0.32, (C): 3.15 +/- 0.43. In all preparations the cobalt content was below the detection limit (less than 5 pmol mg-1 protein).

Animals↗

Manganese absorption in humans: the effect of phytic acid and ascorbic acid in soy formula.

The absorption of manganese from soy formula was studied in adult volunteers by extrinsic labeling of test meals with 54Mn, followed by whole-body retention measurements for approximately 30 d after intake. Eight subjects participated twice in each of the two studies, acting as his or her own control. Soy formula containing the native content of phytic acid was compared with a similar dephytinized formula: geometric mean manganese absorption increased 2.3-fold from 0.7% (range: 0.2-1.1%) to 1.6% (range: 1.0-7.2%) (P < 0.01) with the dephytinized formula. In addition, the effect of the ascorbic acid content of the phytic acid-containing formula was investigated. Manganese absorption was not influenced by an increase in the ascorbic acid from 625 mumol/L (110 mg/L) to 1250 mumol/L (220 mg/L): the geometric mean manganese absorption was 0.6% (range: 0.3-1.0%) and 0.6% (range: 0.3-1.1%), respectively. In conclusion, fractional manganese absorption was approximately doubled by the dephytinization of soy formula but was not influenced by an increase in the ascorbic acid content of a soy formula containing the native amount of phytic acid.

Adult↗

Metabolism of ascorbic acid and ascorbic-2-sulfate in man and the subhuman primate.

Man does not catabolize ascorbate to CO2, whereas the monkey does catabolize ascorbate and ascorbate sulfate to CO2 when these compounds are given orally. However, it takes the same length of time to produce frank scurvy in both man and the monkey, thus indicating that the comparative storage, rate of use, and mode of metabolism of ascorbate is similar in both species. Preliminary feeding and isotope studies conducted on monkeys are in agreement with the fact that only a small amount of labeled ascorbate or ascorbate sulfate equilibrated with body stores. These results are in agreement with published ascorbic acid requirements of 10 mg/kg body weight. In our experiments, 250 mg/day had to be fed to a 10-kg monkey to completely clear all signs of scurvy and return blood ascorbate levels to normal. Ascorbic acid administered intravenously to monkeys appears to equilibrate completely with the ascorbate pool(s). Ascorbate sulfate was found to be a urinary metabolite of both ascorbic-1-14C acid and ascorbic-6-14C acid fed orally to monkeys.

Animals↗

Determination of L-ascorbic acid and total ascorbic acid in vascular and nonvascular plants by capillary zone electrophoresis.

A capillary zone electrophoresis (CZE) procedure for the determination of l-ascorbic acid (AA) and total ascorbic acid (TAA, as the sum of AA and dehydroascorbic acid) in vascular plants, lichens, bryophytes, and liverworts is described. The samples were frozen in liquid nitrogen and extracted with 2% metaphosphoric acid. To determine AA, an aliquot was directly injected in a fused-silica capillary. The determination of TAA was performed upon its reduction with dl-homocysteine at pH 7. The background electrolyte contained 60 mM sodium chloride, 60 mM sodium dihydrogen phosphate, 1 x 10(-4)% hexadimetrine bromide, and NaOH up to pH 7. The procedure was rapid and highly reproducible; the limit of detection was 0.2 microg ml-1, which corresponded to 0.4 mg/100 g of sample. The method was validated by comparison with an enzymatic assay. While the enzymatic assay failed to quantify AA in some plants owing to strong background UV absorption, CZE was successful in all the extracts assayed.

Ascorbic Acid↗

Biologically significant scavenging of the myeloperoxidase-derived oxidant hypochlorous acid by ascorbic acid. Implications for antioxidant protection in the inflamed rheumatoid joint.

Ascorbic acid, at physiological concentrations, can scavenge the myeloperoxidase-derived oxidant hypochlorous acid at rates sufficient to protect alpha 1-antiprotease against inactivation by this molecule. The rapid depletion of ascorbic acid at sites of inflammation, as in the inflamed rheumatoid joint, may therefore facilitate proteolytic damage.

Antioxidants↗

Simultaneous determination of acetaminophen, acetylsalicylic acid and ascorbic acid in tablet form using HPLC.

The purpose of the present study was to develop a simple and accurate HPLC method to measure the amount of each agent in a multidrug pharmaceutical formulation. Three drugs, acetaminophen, acetylsalicylic acid and ascorbic acid, were analyzed simultaneously. A commercial pharmaceutical effervescent tablet was examined and the amount of each of these agents successfully determined. The present method appears to be more convenient than the current procedures described in American and British Pharmacopoeias in which each drug is assayed separately.

Acetaminophen↗

The determination of dehydroascorbic acid and ascorbic acid in the serum and synovial fluid of patients with rheumatoid arthritis (RA).

Using a novel high performance liquid chromatography (HPLC) determination of ascorbic acid and dehydroascorbic acid, we have measured the relative amounts of ascorbate and dehydroascorbate in 20 normal controls and in paired sera and synovial fluid from 13 patients with rheumatoid arthritis (RA). In complete contrast to previous published findings we were able to detect dehydroascorbate in normal human sera (12.0 +/- 3.7 mumol/l), while the mean and range of ascorbate measured in normals was 69.6 +/- 20.6 mumol/l. These findings were completely reversed in rheumatoid sera (21.8 +/- 8.6 mumol/l and 5.1 +/- 5.0 mumol/l for dehydroascorbate and ascorbic acid respectively). In several rheumatoid sera no ascorbate could be detected. In paired synovial fluid and serum samples, there was always more dehydroascorbate detected in synovial fluids than in the corresponding sera (p less than 0.01). The data suggests that the reduced level of ascorbate and increased level of dehydroascorbate may be a reflection of the increased antioxidant and free-radical scavenging activity of the vitamin in RA, especially within the inflamed joint.

Adult↗

Comparison of absorption of erythorbic acid and ascorbic acid in guinea pig small intestine.

Both the ascorbic acid (AsA) and erythorbic acid (ErA) absorption in the small intestine of guinea pigs were determined by the perfusion of the small intestine using isotonic phosphate buffer recycled in situ. The absorption rate of AsA in the small intestine of guinea pigs was higher than that of ErA; however, Km of AsA absorption was lower than that of ErA in normal guinea pigs. In AsA-deficient guinea pigs, the absorption rates of both AsA and ErA were higher than those in normal ones. The absorption of AsA and ErA in the small intestine of guinea pigs was inhibited by ouabain. Furthermore, AsA and ErA inhibited each other's absorption. Based on the results, the net amount of the absorbed ErA in the small intestine may be lower than that of AsA, and ErA absorption mechanism seemed to be similar to that of AsA. The absorption rate of both AsA and ErA in the small intestine of guinea pig might be dependent on the AsA level in the tissues.

Animals↗