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Sex variation in ascorbic acid catabolism.

Male and female albino rats of same age and body weight were pair fed with laboratory stock diet and ascorbic acid, dehydroascorbic acid and diketogulonic acid were determined in the liver and urine, while in blood only ascorbic acid was estimated. Male rats had concentration higher of ascorbic acid in liver and urine as compared with females, while there were no significant variations in the contents of dehydroascorbic acid and diketogulonic acid. Hepatic and renal 2, 3-diketoaldonate decarboxylase, and hepatic dehydroascorbatase were also found to be significantly higher in male rats. Similar sex variations were also observed in ascorbic acid catabolism in guinea pigs without any differences in urinary ascorbic acid contents.

Animals

Manganese catalysis of dopamine oxidation.

Manganese catalysis of the oxidation of dopamine by air was studied as part of an investigation of possible manganese intoxication amongst a group of Aborigines living on manganese-rich soil on Groote Eylandt, in the Northern Territory of Australia. Manganese significantly increased the oxidation rate of dopamine, and the manganese complexes with some purines were especially efficient catalysts. An oxidation mechanism, involving a manganese(II)/(III) redox couple and a semiquinone free radical intermediate, is proposed. Stoichiometric hydrogen peroxide was produced by the oxidation, and the oxidation products of dopamine were highly toxic to the marine diatom Nitzschia closterium. Hydrogen peroxide and the superoxide radical did not oxidize dopamine at physiological pH. Some electrophilic compounds, including ascorbic acid, dehydroascorbic acid and thiamine, effectively inhibited dopamine oxidation. The Groote Eylandt Aborigines are likely to be deficient in ascorbic acid (vitamin C) and thiamine (vitamin B1), and these deficiencies, as well as their lifestyle, may predispose them to manganese intoxication.

Catalysis

The influence of irradiation and packaging on the quality of prepacked vegetables.

During storage, transport and sale of prepacked vegetables, their quality deteriorates very quickly by microbiological decay, discolouration and desiccation. The experiments described were carried out with endive (Chichorium endiva L.) as a representative of the leaf vegetables. After processing (cutting, washing and drying), the product was packed in polythene bags without and with 4 perforations of 2 mm diameter. The endive was irradiated with 0 and 100 krad gammarays and afterwards stored at 10 degrees C, thus simulating the transport and sale temperature. The quality was studied on the basis of the following parameters. The gas composition in the bags, the microbiological composition, the Vitamin C (Ascorbic acid + Dehydroascorbic acid) content and the sensory quality. The experiments showed that the modified gas composition in the non-perforated bags, caused by the respiratory activity of the cut product, improved the quality. The low O2 and high CO2 content prevented discolouration, retarded the senescence and diminished the Vitamin C losses. The initial Total Viable Count of endive was 10(6) to 10(7) and the number of Enterobacteriaceae amounted to approx. 10(5) g-1 product, and increased in a couple of days to 10(8) and 10(7) g-1 respectively, resulting in decay of the product. The contamination with aerobic and anaerobic spores was low, viz. 10(1) to 10(2) g-1 and did not increase at 10 degrees C. An irradiation treatment of 100 krad reduced the Total Viable Count and the Enterobacteriaceae with 3-4 decimals, by which the shelf life increased with about 100% and almost all the Enterobacteriaceae were eliminated, so that the hygienic quality was improved. In prepacked endive nearly half of the Total Vitamin C content was lost within two days. The retention was strongly affected by the presence or absence of perforations; the effect of an irradiation treatment was slight.

Enterobacteriaceae

[On the mechanism of ascorbic acid induced methemoglobin reduction of human erythrocytes (author's transl)].

Ascorbic acid and dehydroascorbic acid penetrate the human erythrocyte membrane. In vitro methemoglobin is reduced nonenzymatically by both substances in concentrations of 10(-2) M to 10(-3) M. Dehydroascorbic acid is reduced nonenzymatically to ascorbic acid by GSH, even with low GSH-content of erythrocytes. Under physiological conditions ascorbic acid induced methemoglobin reduction is far less important than reduction by the NADH dependent methemoglobin reductase system. In methemoglobinemic conditions caused by toxic effects or by congenital methemoglobin reductase deficiency treatment with ascorbic acid is possible. However, critically increased methemoglobin content of the blood higher than 30% makes therapy with methylene blue necessary.

Ascorbic Acid

Assessment of human vitamin C status.

Since no reliable functional markers of human vitamin C status have been demonstrated, determination of vitamin C levels in blood plasma and/or leukocytes remains the current choice for individual and population assessments. Newer analytical techniques, especially high-performance liquid chromatography, allow determination of reduced (ascorbic acid), oxidized (dehydroascorbic acid), or total amounts of vitamin C in biological specimens or foods. Plasma levels of vitamin C forms are easily determined but may not reflect tissue content as well as leukocyte levels. The vitamin C content of leukocyte cell types varies severalfold and, unlike plasma, leukocytes may contain an appreciable fraction of dehydroascorbic acid. The effects of sex, age, cigarette smoking, drugs, and physiological factors on vitamin C levels are better known for plasma than leukocytes. To realize the potential of leukocytes as measures of vitamin C status, continued work is needed in standardizing the methodology and interpretive guidelines and simplifying the technique for blood processing. The search for specific functional markers of vitamin C deficiency should continue. Candidate markers may involve pathways of carnitine or collagen metabolism, immunocompetence, or antioxidant defense.

Ascorbic Acid

Ascorbate- and dehydroascorbic acid-mediated reduction of free radicals in the human erythrocyte.

Nitroxides were used as models of persistent free radicals to study the antioxidant function of ascorbic acid in the human erythrocyte. It was concluded that: 1) ascorbate and other reductant(s) derived from dehydroascorbic acid (DHA) in the presence of thiols are the only significant reducing agents for nitroxides, 2) glutathione and DHA reduce nitroxides by a process that cannot be inhibited by ascorbic acid oxidase, 3) erythrocytes can be depleted of ascorbic acid by exhaustive washing in the presence of membrane-permeable cationic nitroxides such as N,N-dimethylamino-Tempo, 4) ascorbate-depleted cells do not reduce nitroxides; however, nitroxide reduction is restored when the cells are incubated with DHA, 5) reduction of nitroxides in ascorbate-depleted, DHA-treated cells is significantly faster than in buffered solutions of DHA and glutathione, 6) several equivalents of nitroxide are reduced relative to the intracellular ascorbate pool, 7) sustained nitroxide reduction is observed even when most of the intracellular ascorbate is oxidized, 8) spin trapping of oxyradicals in tert-butyl hydroperoxide-treated cells is accelerated with ascorbate depletion and inhibited with ascorbate loading, 9) ascorbate can be quantified within intact cells by analyzing the initial reduction rates of membrane-permeable cationic nitroxides, and 10) DHA-stimulated reduction of cationic nitroxides is slower and less extensive in erythrocytes deficient in glucose-6-phosphate dehydrogenase than in normal erythrocytes.

Ascorbic Acid

Active transport of ascorbic acid across the retinal pigment epithelium of the bullfrog.

Known functions of the RPE include glucose, water and retinoid transports; an ion transport mechanism utilizing a Na(+)-K(+)-ATPase pump located in the apical membrane has been proposed. Recent studies with cultured RPE cells of cat and bovine indicate that the RPE takes up ascorbate by an active mechanism. In this study we use a mounted bullfrog RPE preparation to study unidirectional and net fluxes of radiolabeled (14C)-ascorbic acid (AA), (14C)-dehydroascorbic acid, (3H)-L-glucose(L-glu) and (14C)-3-O-methyl-D-glucose(mD-glu) in an effort to explore the mechanism whereby AA moves across this tissue. Comparative flux studies with AA indicated that the retina to blood side (apical to basal:AB) flux of AA was more than 6x that of L-glu, a passive marker of comparable size. The reverse BA flux of AA was not significantly different from that of L-glu. Flux studies of L-glu, mD-glu and dehydroascorbic acid revealed no "net" flux across the mounted RPE; significantly, only AA demonstrated a net flux from retina to choroid (AB). The AB flux of reduced ascorbate was significantly greater than that of dehydroascorbic acid indicating specificity of carrier mediation. Apical ouabain (10(-4) M) and sodium replacement in the bathing medium reduced the AB and net flux of AA significantly suggesting the requirement of a functioning Na(+)-K(+)-ATPase on the apical side membrane of the RPE. Energy blocker, dinitrophenol decreased unidirectional AB and net AA fluxes.(ABSTRACT TRUNCATED AT 250 WORDS)

3-O-Methylglucose

Compartmentation of redox metabolites in the anterior eye segment?

In bovine corneal epithelium, stroma, and aqueous humor the levels of ascorbic acid (ASC) and dehydroascorbic acid (DHA) were investigated. Two methods were used, the photometric assay with 2,6-dichlorophenolindophenol and the formation of osazone by 2,4-dinitrophenylhydrazine. The ASC levels in the corneal epithelium and aqueous humor were found to be in the millimolar range, the ASC/DHA ratio being about 10. The stromal ASC and DHA levels were much lower, with a ratio of 0.7. ASC and DHA had similar levels and ratios to those of reduced and oxidized glutathione (GSH/GSSG) reported in the literature. In the corneal epithelium the redox ratio of glutathione was higher than that of ascorbic acid. Therefore, glutathione was supposed to reduce dehydroascorbic acid.

Animals

Effect of species differences and dietary vitamin C on the concentration of ascorbate- and acid-soluble thiol in fish eye.

Data presented confirm the essentiality of modification of the dinitrophenylhydrazine (DNPH) method to analyze the total ascorbic acid and dehydroascorbic acid in ocular tissues and stress the need of corrections for the interfering substances. Variations in ascorbate and thiol concentrations in the lens, retina and aqueous humour of freshwater fish belonging to the Cyprinidae family were examined. The interspecific variability of ascorbate concentration was highest in the aqueous humour and lowest in the retina. The high ascorbate concentration in the retina seems to reflect the importance of the sense of vision in fish life-style as compared to chemo- and acoustico lateralis senses. The regional distribution of the total ascorbate is in the order of decreasing concentrations: retina, lens and aqueous humour. However, the retinal ascorbate is almost exclusively in the oxidized form, and the lenticular ascorbate is almost exclusively in the reduced form. Thiol concentration in the lens is five- to tenfold that in the retina and aqueous humour. This explains the oxidation status of ascorbate in different eye compartments of the eye. After 30 days on diets containing various levels of ascorbic acid or ascorbic acid sulphate, the ascorbate concentration in the eye compartments of common carp (Cyprinus carpio L.) was determined. Ocular tissue can be used to monitor the development of the ascorbate status in fish, and the retina is the most responsive tissue to the enhanced or depleted ascorbate levels.

Animals

Further investigations into the relationship between the dopaminergic system, ascorbic acid and uric acid in the rat striatum.

Levels of dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), ascorbic acid (AA), dehydroascorbic acid (DHAA), and uric acid were determined in the rat striatum following single apomorphine (1 mg/kg), scopolamine (0.6 mg/kg), pilocarpine (4 mg/kg), or pilocarpine + scopolamine (4 and 0.6 mg/kg, respectively) injections. The decrease in DOPAC levels and in the DOPAC/DA ratio, induced by the pharmacological manipulation, was linearly correlated with the increase in DHAA levels (r = -0.9060, P less than 0.05) and with the increase in the DHAA/AA ratio (r = -0.9004, P less than 0.05), respectively. It is concluded that dopaminergic activation or cholinergic inhibition both increase striatal AA oxidation, which is correlated with a decrease in DA turnover.

3,4-Dihydroxyphenylacetic Acid

Effect of organic mercurials and sulfhydryl compounds on the urease activity of Proteus: inhibition by urine and ascorbic acid.

Meralluride, mercaptomerin, ethacrynic acid, and penicillamine inhibited urease activity of Proteus mirabilis. The activity of the organic mercurials and ethacrynic acid was markedly inhibited by human and dog urine. Antiurease activity could not be detected in the urine of a human and a dog given meralluride by injection. Urine from patients receiving penicillamine also failed to inhibit urease activity. Ascorbic acid inhibited, whereas dehydroascorbic acid enhanced, the activity of the mercurials, but neither agent altered the inhibitory effect of urine. The lethal effect of meralluride against Proteus occurred at the same concentration at which urease activity was inhibited, but penicillamine inhibited the enzymatic activity without affecting viability of the organism. The data suggest that these sulfhydryl-reactive compounds will not be useful against Proteus infections of the urinary tract.

Animals

Ascorbic acid status in uremics.

Plasma levels of ascorbic acid (AA) and dehydroascorbic acid (DHA) were estimated in 27 patients of end stage renal failure (ESRF) on standard conservative therapy (group A) and 9 patients of ESRF on maintenance haemodialysis (MHD; group B). Fourteen healthy subjects matched for age and sex served as control (group C). The dietary intake of vitamin C was significantly decreased in group A than in group B compared to control. Similarly, plasma AA was significantly lowered to 0.801 +/- 0.283 mg per cent in group A compared to 1.421 +/- 0.47 mg per cent in control. While it was just lowered to 1.058 +/- 0.272 mg per cent in group B. Although plasma level of DHA was raised to 0.243 +/- 0.486 mg per cent and 0.166 +/- 0.54 mg per cent in groups A and B respectively, the increase was not statistically significant. In our present study, the DHA/AA ratio was found to be inversely proportional to the plasma AA. Further, this ratio has been claimed to be a better indicator of overall reducing atmosphere (i.e., profile of vitamin C) of the body.

Ascorbic Acid

Ascorbic acid and diabetes mellitus.

Dehydroascorbic acid, the oxidized form of vitamin C, causes diabetes when injected into animals and has been reported to be present in increased amounts in the blood of patients with diabetes mellitus and even in prediabetics. One of the earliest changes in diabetes mellitus is electron-microscopic evidence of damage to the inner endothelial lining of the blood vessels. Certain bioflavonoids, which are natural non-toxic food substances from plants, like rutin from buckwheat, prevent the oxidation of ascorbic acid and seem to protect the endothelium when given with vitamin C; it is therefore suggested that all vitamin C tablets should be combined with these flavonoids.

Animals

Reduction of extracellular methemoglobin by erythrocytes.

Erythrocytes, suspended in a glucose-containing buffer, catalyzed the partial reduction of extracellular methemoglobin. Physiological concentrations of ascorbic acid or dehydroascorbic acid greatly enhanced the rate of reaction and the ultimate extent of reduction. The relationship between erythrocyte concentration and initial reaction rate was nonlinear, which suggested that the rate limiting factor was not an erythrocyte membrane enzyme. Also, significant dehydroascorbate-stimulated reduction occurred even when the erythrocytes and methemoglobin were separated by a dialysis membrane. The above observations indicate that the transfer of reducing equivalents across the erythrocyte membrane and reduction of extracellular methemoglobin can be accomplished by release and recycling of ascorbic acid.

Ascorbic Acid

Analysis of water-soluble vitamins in total parenteral nutrition solution by high pressure liquid chromatography.

A stability-indicating high pressure liquid chromatographic method was developed to determine the stability of water-soluble vitamins in total parenteral nutrition mixtures. Folic acid and thiamine were determined by direct injection and precolumn enrichment, followed by UV detection. Nicotinamide and pyridoxine were determined simultaneously without sample pretreatment by UV detection of nicotinamide and fluorescence detection of pyridoxine. Riboflavin 5'-phosphate was also determined without elaborate sample handling and by fluorescence detection. Ascorbic acid was determined as the sum of ascorbic acid and dehydroascorbic acid. After derivatization to a quinoxaline the latter substance was determined by direct injection and fluorescence detection.

Ascorbic Acid

Effects of cortical ablation on apomorphine- and scopolamine-induced changes in dopamine turnover and ascorbic acid catabolism in the rat striatum.

Levels of dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), ascorbic acid and dehydroascorbic acid (DHAA) were measured by HPLC in the striatum of rats whose fronto-parietal cortex had been unilaterally ablated after a single injection of apomorphine (1 mg/kg s.c.), scopolamine (0.6 mg/kg s.c.) or L-glutamate (500 mg/kg i.p.). Unilateral cortical ablation decreased striatal levels of glutamate in both striata ipsilateral (35%) and contralateral (17-25%) to the lesion. Apomorphine and scopolamine significantly increased (+94 and +122%, respectively) the DHAA/ascorbic acid ratio in the striata ipsilateral to the lesion in unoperated and sham-operated rats (+72 and +34%, respectively), but both drugs failed to increase it in ablated rats. L-Glutamate significantly increased the DHAA/ascorbic acid ratio in unoperated (+53%) and ablated rats (+37%). The increase in sham-operated rats (+34%) did not reach statistical significance. Apomorphine and scopolamine significantly decreased the DOPAC/DA ratio in the striata ipsilateral to the lesion of unoperated, sham-operated and ablated rats. The decrease in the DOPAC/DA ratio induced by apomorphine and scopolamine was greater in ablated rats than in sham-operated rats. L-Glutamate induced only minor changes in striatal DA and DOPAC levels. We conclude that the apomorphine- and scopolamine-induced increase in ascorbic acid oxidation in the striatum requires intact cortico-striatal glutamatergic pathways. Cortical ablation potentiates the apomorphine- and scopolamine-induced inhibition of striatal DA turnover.

3,4-Dihydroxyphenylacetic Acid

Vitamin C and vitamin E status in the spontaneously diabetic BB rat before the onset of diabetes.

Ascorbic acid (AA), dehydroascorbic acid (DHAA), and vitamin E were measured in tissues and plasma of 30 control and 30 spontaneously diabetic BioBreeding rats (BBdp) during development and before the onset of diabetes. At weaning, rats were fed an AIN-76 semisynthetic diet for 30, 64, or 113 days, after which plasma and tissues from 10 rats of each group were collected and analysed for AA, DHAA, and vitamin E. AA and DHAA levels were significantly increased in plasma and spleen of the diabetes-prone rats compared with those of the control group at 30 and 64 days, but the difference disappeared by 113 days. No differences were observed in liver, adrenals, thymus, and pancreas at any of the time periods. However, lower levels of vitamin E were observed in adrenal gland, thymus, and pancreas of the diabetes-prone rats. It is concluded that BBdp rats have an altered metabolism of AA, DHAA, and vitamin E, before the onset of diabetes. These changes could be due to genetic and physiological factors operating during development of this rat strain.

Adrenal Glands

2-Imidazolethiones protect ascorbic acid from oxidation induced by copper.

The oxidation of ascorbic acid to dehydroascorbic acid was accelerated by metal ions such as copper. This stimulation of ascorbate oxidation was inhibited by the addition of 2-imidazolethiones and other sulfhydryl-containing compounds, but not by 2-imidazolones or phenytoin. Although the 2-imidazolethiones interacted with copper, as shown by a decrease in the ultraviolet absorbance of the compounds, the product formed still protected ascorbate from oxidation. The 2-imidazolethiones are proposed to complex copper through their free -SH groups.

Ascorbic Acid