ascorbic-acid and deoxypyridinoline

Vitamin C is an essential micronutrient. Absence from the diet will result in the deficiency disease scurvy, typically characterised by weakening of collagenous structures. High intakes of vitamin C have been associated with decreased incidence or severity of a number of diseases, including cancer and cardiovascular disease. These beneficial effects may be attributed to its antioxidant properties, although the exact mechanisms of action remain elusive. It is also unclear what intake levels are required for optimal health benefits. The task of defining optimal intakes is hindered by the lack of a reliable functional marker of tissue vitamin C status in man. Many different pathways have been investigated, but none of them have measurable outcome variables relating directly to scorbutic changes. The bone-collagen formation pathway has the potential to provide a functional index of tissue vitamin C adequacy. Vitamin C acts as a cofactor for the enzyme lysyl hydroxylase, which is required for the hydroxylation of lysine residues in procollagen chains. Pyridinoline is a mature collagen cross-link formed from three hydroxylysine residues, deoxypyridinoline is formed from two hydroxylysine and one lysine residue. Guinea-pig studies have shown an alteration in the pyridinium cross-link ratios in response to graded vitamin C intakes (Tsuchiya & Bates, 1998). In order to investigate whether these changes can be seen in a human population group, a study was carried out in rural Gambia, where there is a marked seasonal variation in dietary vitamin C. The present review discusses the rationale behind the study and presents some preliminary results.

Topics: Amino Acids; Antioxidants; Ascorbic Acid; Ascorbic Acid Deficiency; Bone and Bones; Collagen; Humans; Hydroxylation; Seasons

1. This study examines whether the supplementation of isoflavones (ISO) exerts beneficial effects on serum and urinary biomarkers of bone metabolism. 2. A total of 102 women were randomly assigned to three groups taking either ISO (40 mg) tablets, tablets containing vitamins C (25 mg) and E (5 mg) (V) or placebo tablets (vehicle only) (P) daily for 4 weeks, in a double-blind parallel placebo controlled design. 3. Among the 67 women who completed the study (24 on ISO, 24 on V, 19 on P), only ISO tablets were proven to decrease significantly urinary deoxypyridinoline (Dpd) excretion (P < 0.05 vs before), a specific biomarker of bone resorption, but there was no significant difference in serum bone gamma-carboxyglutamic acid-containing protein (BGP), a specific serum biomarker of bone formation. 4. Among the 67, 25 women were postmenopausal (8 on ISO, 12 on V, 8 on P) and only ISO tablets decreased significantly urinary Dpd excretion (P < 0.05 vs before) in them. The reduction rate of Dpd in ISO group was also significantly greater than that in P group (P < 0.01). 5. Dietary supplementation of vitamins C 25 mg and E 5 mg did not affect urinary Dpd.

Topics: Adult; Amino Acids; Antioxidants; Ascorbic Acid; Asian People; Biomarkers; Bone and Bones; Bone Resorption; Double-Blind Method; Female; Glycine max; Humans; Isoflavones; Middle Aged; Osteocalcin; Postmenopause; Premenopause; Vitamin E

Prelabor rupture of the fetal membranes affects approximately 10% of women at term, resulting in an increased risk of maternal and neonatal infection. Evidence suggests that membrane rupture is related to biochemical processes involving the extracellular matrix of the membranes. We tested the hypothesis that prelabor ruptured membranes are characterized by reduced collagen concentrations, altered collagen cross-link profiles, and increased concentrations of biomarkers of oxidative damage. We also set out to determine whether these effects are modulated by ascorbic acid status. In a case-control study, we explored the role that ascorbic acid, oxidative stress, collagen, and collagen cross-links play in determining membrane integrity and developed a functional assay to assess membrane proteolytic susceptibility. Prelabor ruptured membrane had a reduced ascorbic acid concentration in comparison with controls while protein carbonyl and malondialdehyde concentrations were increased. Collagen concentrations were also reduced in prelabor ruptured membrane, and while the concentration of collagen cross-links was not significantly different between prelabor and timely ruptured membrane, there was a regional variation in cross-link ratio within the amniotic sac. Proteolytic resistance in vitro was reduced in prelabor ruptured membrane and also exhibited regional variation within the amniotic sac. Our findings are strongly supportive of a role for the enhanced degradation of membrane collagen in the determination of prelabor rupture of fetal membranes. The formation of the rupture initiation site is a function of a regional variation in collagen cross-link ratio. Tissue ascorbic acid status may be an important mediator of these processes.

Topics: Adult; Amino Acids; Ascorbic Acid; Case-Control Studies; Collagen; Disease Susceptibility; Extraembryonic Membranes; Female; Fetal Membranes, Premature Rupture; Humans; Hydroxyproline; Infant, Newborn; Malondialdehyde; Oxidative Stress; Pregnancy; Proteins; Reference Values

Vitamin C (ascorbate) is essential for hydroxylation of prolyl and lysyl residues in nascent collagen, the failure of which leads to connective tissue lesions of scurvy. Of the pyridinium-type cross-links in mature collagen, pyridinoline requires more hydroxylysyl residues than does deoxypyridinoline. Our study tested the hypothesis that pyridinoline:deoxypyridinoline ratios in urinary degradation products may vary with ascorbate status in man. These ratios were compared between British and Gambian prepubertal boys, mean age 8.3 years, and in Gambian boys between two seasons with contrasting ascorbate availability. The mean cross-links ratio in 216 British boys was 4.36 (SD 0.71), significantly greater (P<0.0001) than in sixty-two Gambian boys: 3.83 (SD 0.52). In the Gambians the cross-links ratio was significantly higher in the dry season (with high ascorbate intake and status) than in the rains (with low intake and status). A 7-week controlled intervention was carried out in Gambian boys during the rainy season (the 'hungry' season, when vitamin C-containing foods are virtually unavailable): 100 mg ascorbate/d was given to one group of thirty-two Gambian boys and placebo to another group. The intervention did not, however, significantly alter the cross-link ratio, possibly because the response time and/or intervention-response delay is >7 weeks. If confirmed, the putative association between ascorbate and collagen cross-link ratios in man could become the basis for a functional test for adequacy of ascorbate status.

Topics: Amino Acids; Anthropometry; Ascorbic Acid; Ascorbic Acid Deficiency; Biomarkers; Body Mass Index; Bone and Bones; Child; Child, Preschool; Collagen; Creatinine; Gambia; Humans; Male; Rain; Seasons; United Kingdom

Mild-to-moderate vitamin C depletion in weanling guinea-pigs affects pyridinoline:deoxypyridinoline (collagen cross-link) ratios in femur shaft and urine, attributed to impairment of hydroxylation of collagen lysine. We investigated: (1). whether the picture at two time points is compatible with progressive accumulation of abnormal collagen; (2). whether any changes are seen in skin, where little deoxypyridinoline occurs; (3). whether total food restriction has similar effects. Male weanling Dunkin-Hartley guinea-pigs were fed diets containing either 0.5 (vitamin C-restricted) or 160.0-320.0 (vitamin C-adequate) mg vitamin C/d. Two groups receiving the vitamin C-adequate diet received it ad libitum. Two other groups received the vitamin C-adequate diet in a restricted amount, limited to that which permitted nearly the same growth rate as in the vitamin C-restricted groups. Animals were fed for 4 or 8 weeks; urine was collected, and vitamin C and collagen indices were measured. In the femur shaft, the hydroxyproline content per unit weight was unaffected by vitamin C restriction or by total food restriction. Deoxypyridinoline was increased and the pyridinoline:deoxypyridinoline ratio was decreased in vitamin C-restricted groups, but not in food-restricted groups. Changes in the value of the ratio were greater after 8 than after 4 weeks. Urine indices mirrored bone indices. In skin, the main effect of vitamin C restriction was to reduce hydroxyproline. Here, the cross-link ratios changed less markedly than in bone, and there was less deoxypyridinoline. We conclude that the picture at two time points is compatible with a progressive accumulation of pyridinoline-enriched collagen in vitamin C-deprived animals, that the picture in skin differs from that of bone and urine, and that cross-link changes are not produced by total food restriction.

Topics: Amino Acids; Animals; Ascorbic Acid; Ascorbic Acid Deficiency; Body Weight; Bone and Bones; Collagen; Eating; Food Deprivation; Guinea Pigs; Male; Skin

Ehlers-Danlos syndrome type VI (EDS VI) is an autosomal recessive disorder of connective tissue characterized by hyperextensible, friable skin and joint hypermobility. Severe scoliosis and ocular fragility are present in some patients. This disease is caused by defective collagen lsyl hydroxylase, a vitamin C-dependent enzyme that converts lysyl residues to hydroxylysine on procollagen peptides. Hydroxylysine is essential for the formation of the covalent pyridinium cross-links pyridinoline (Pyr) and deoxypyridinoline (Dpyr), among mature collagen molecules. Pyr derives from three hydroxylysyl residues, whereas Dpyr derives from one lysyl and two hydroxylysyl residues. Patients with EDS VI have high urinary excretion of Dpyr, resulting in a high ratio of Dpyr-Pyr. In this study, we evaluate content and production of pyridinium cross-links in the skin and cultured fibroblasts from patients with EDS VI. The skin of normal controls contained both Pyr and Dpyr, with a marked predominance of Pyr as observed in normal urine. The skin of patients with EDS VI had reduced total content of pyridinium cross-links, with the presence of Dpyr but not Pyr. Long-term cultures of control fibroblasts produced both Pyr and Dpyr, with a pattern resembling that of normal skin. By contrast, cross-links were not detected in dermal fibroblasts cultured from patients with EDS VI. Vitamin C, which improves the clinical manifestations of some patients with EDS VI, decreased Dpyr accumulation though only minimally affecting Pyr content in control cells. By contrast, addition of vitamin C to fibroblasts from patients with EDS VI stimulated the formation of Dpyr more than that of Pyr and greatly increased total pyridinium cross-link formation. These results indicate that qualitative and quantitative alterations of pyridinium cross-links occur in skin and in cultured dermal fibroblasts of patients with EDS VI and may be responsible for their abnormal skin findings. The vitamin C-stimulated production of Dpyr and Pyr in fibroblasts from patients with EDS VI may explain at least in part the therapeutic effects of this vitamin in EDS VI.

Topics: Adolescent; Amino Acids; Ascorbic Acid; Cells, Cultured; Child; Collagen; Cross-Linking Reagents; Ehlers-Danlos Syndrome; Female; Fibroblasts; Humans; Hydroxylysine; Lysine; Pyridinium Compounds; Skin

The purpose of this study was, first to explore metabolic interactions between Cu and ascorbic acid in guinea-pigs, particularly with respect to any possible disadvantages of high ascorbate in the presence of low Cu intakes, and second, to test the hypothesis that variations in ascorbate and/or Cu status might influence collagen cross-linking, either by inducing a change in the crosslinks: hydroxyproline ratio, or by inducing a change in the pyridinoline: deoxypyridinoline crosslinks ratio. Four matched groups, each of eight male weanling Dunkin-Hartley guinea-pigs, were maintained on purified diets containing either no added Cu, or 150 mg Cu/kg diet, and either 0.1 g or 30 g ascorbic acid/kg diet. They were then killed 8 weeks later, and the following indices were measured: body and organ weights; blood haemoglobin; adrenal ascorbate concentrations; Cu concentrations in plasma, liver and femur; superoxide dismutase (EC 1.15.1.1) activity in whole blood and liver; hydroxyproline, pyridinoline and deoxypridinoline in femur and in urine. The principal observations were: Cu intake significantly affected blood and tissue Cu concentrations and superoxide dismutase activity; and ascorbic acid intake significantly affected adrenal ascorbate levels and the deoxypyridinoline: pyridinoline cross-links ratio, especially in bone (femur). There was evidence of a significant interaction between ascorbate and Cu with respect to adrenal and plasma Cu concentrations, blood superoxide dismutase activity and body weights. We conclude that interactions between ascorbate and Cu at the functional level were present but modest, and that a new and potentially powerful functional index of ascorbate status may exist within the deoxypyridinoline:pyridinoline collagen cross-link ratio.

Topics: Adrenal Glands; Amino Acids; Analysis of Variance; Animals; Ascorbic Acid; Bone and Bones; Copper; Diet; Guinea Pigs; Hemoglobins; Liver; Male; Superoxide Dismutase