The experiments in this thesis tested <i>in vitro</i> and <i>in vivo</i> the proposal that zinc-deficient superoxide dismutase, resulting from mutations or oxidative damage to the enzyme, gains ascorbate oxidase activity that contributes to the pathology of amyotrophic lateral sclerosis (ALS). They also tested whether flavonoids can help protect against this activity.<p>The <i>in vitro</i> experiments showed that zinc-extracted Cu/Zn-SOD (Cu-SOD) as well as SOD treated with H2O2 or H2O2 plus ascorbate accelerated ascorbate oxidation 100 to 300 %, while native SOD had no effect. With Cu-SOD, the activity was unaffected by EDTA, EGTA, or catalase, showing that the catalytic copper was firmly bound and that the H2O2 product of SOD activity was not responsible. Catechin and uric acid slowed ascorbate oxidation by Cu-SOD by 72% and 67%, respectively.<p>The <i>in vivo</i> study investigated tissue levels of ascorbate and biomarkers of oxidative stress in a transgenic mice bearing a mutation in Cu/Zn-SOD as a model of familial ALS (FALS mice), and the effects of dietary ascorbate and quercetin. In FALS mice on control modified AIN93G diet for 10 weeks compared to the wild-type, liver thiobarbituric acid reactive substances (TBARS) were 47% higher and liver oxidized vitamin C was 2800% higher. These results support, in liver, that mutant SOD acquired ascorbate oxidase activity and increased oxidative stress. The only difference in other tissues was a 136% increase in GSH/GSSG ratio in thigh muscle of FALS mice.<p>In dietary treatments of FALS mice, spinal cord TBARS was 93 % higher with ascorbate-supplemented diet compared to control diet, suggesting that dietary ascorbate increased oxidative stress. Also in spinal cord, oxidized-vitamin C was 250% higher in ascorbate + quercetin-fed FALS mice, which suggests there is no protection by quercetin against ascorbate oxidation. In brain, protein thiols were 56% and 58% lower in quercetin-fed and ascorbate + quercetin-fed FALS mice, suggesting that quercetin worsened oxidative damage. In liver, quercetin feeding produced a 40% decrease in vitamin C, total vitamin C and oxidized-vitamin C, perhaps by down-regulating ascorbate biosynthesis. Overall the results support a gain of ascorbate oxidase activity of mutant SOD in ALS, but do not support protection by dietary treatment with ascorbate or quercetin.

Zugl.: Zürich, Univ., Diss., 2008

Thesis (M. Phil.)--University of Hong Kong, 1983.

No description available.

No description available.

No description available.

A reverse-phase high pressure liquid chromatographic method for the detection and quantitation of unchanged ascorbic acid in human urine is described. Twenty-four hour ascorbate excretion profiles from nine subjects were determined. The urinary excretion profiles for orally ingested and intravenously infused ascorbic acid in these nine subjects are compared.

Ascorbic acid has been found to promote nitrosothiol decomposition via two pathways. In the first, ascorbic acid acts as a reducing agent for added or adventitious copper (II), producing copper (I). This reacts with the nitrosothiol, giving nitric oxide and disulfide as the ultimate products. The reaction requires only small quantities of ascorbic acid, and is catalytic in copper. The second pathway requires higher concentrations of ascorbic acid, the stoichiometry being one mole of ascorbic acid to two moles of nitrosothiol. The products are nitric oxide and thiol, and the reaction has been interpreted in terms of rate limiting nucleophihc attack by ascorbate at the nitroso nitrogen, followed by decomposition of the 0-nitroso ascorbate formed to nitric oxide and dehydroascorbic acid. The rate equation is first order in both the nitrosothiol and ascorbic acid, and the entropy of activation is significantly negative. pH - rate profiles reveal the ascorbate dianion is much more reactive than the monoanion, and that the neutral form has negligible reactivity. Nitrosation of thione-containing nitrogen heterocycles by nitrous acid leads to the equilibrium formation of =SNO(^+) species; large equilibrium constants are observed. The reactions exhibit many of the features generally observed in nitrosation, including catalysis by halides and thiocyanate, and some participation by dinitrogen trioxide as a nitrosating agent. The nitrosation rate constants are large, approaching values representing the encounter-controlled limit. The =SNO(^+) species are generally unstable, decomposing under acidic conditions to nitric oxide and a disulfide. Decomposition of S-nitrosated 4-thiopyridine showed hydrolysis occurs at pH 7.4, re-forming the thione. The nitroso species reacts rapidly with ascorbate, forming nitric oxide and thione.

Commercially bottled, vitamin C-fortified, cranberry juice cocktail and cranberry-apple (cranapple) juice drinks were stored at 5°C under simulated home-use conditions. Ascorbic acid (AA) and dehydroascorbic acid (DHA) levels were evaluated over a period of seven days by high performance liquid chromatography (HPLC). Total vitamin C content was also determined by a classical spectrophotometric method. The HPLC data revealed a significant decrease of AA during the storage of both juice drinks resulting in losses of 35% and 27% in the cranberry and cranapple drinks, respectively. The loss of ascorbic acid was offset by the simultaneous increase of DHA (a biologically active vitamin) so that the sum of AA+DHA decreased 13% and 0% in the cranberry and cranapple drinks, respectively. Thus ascorbic acid retention was apparently influenced by the nature of the juice system. The spectrophotometric results indicated no significant loss of total vitamin C activity during the storage of either juice drinks. The observed differences between the spectrophotometric and HPLC results were attributed to interference problems associated with the spectrophotometric method. / Graduation date: 1988

Graduation date: 1943