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On the possible involvement of ascorbic acid and copper proteins in leukemia. III. ESR investigations on the interaction between ascorbic acid and some transition metal ions.

The interaction between lyophilized samples of ascorbic acid and Cu2+, Fe3+ or Mn2+ has been investigated by means of ESR spectroscopy. All of the three transition metal ions form complexes with vitamin C, but only in the case of Cu2+ and Fe3+ the interaction results in a reduction of the metal ions. Cu2+ and ascorbic acid seem to form 2 : 1 complexes with an equilibrium constant of about K = 1 X 10(7) mol-1. None of these metal ion complexes exhibits, however, the ESR spectrum obtained with leukemic blood.

Ascorbic Acid

The influence of amino acid source on the stability of ascorbic acid in TPN mixtures.

This study was undertaken to investigate the stability of ascorbic acid and its primary degradation product, dehydroascorbic acid, in total parenteral nutrition (TPN) mixtures. The influence of the type of bag and the commercial source of amino acid on ascorbate degradation was examined, using a stability-indicating high-pressure liquid chromatography (HPLC) method. Ascorbic acid was most stable in multilayered bags, compared with ethylvinyl acetate (EVA) bags. Results indicated that, in multilayered bags, the initial rapid ascorbic acid degradation was greatest in TPN mixtures containing amino acid infusions without reducing activity. In contrast, degradation in TPN mixtures containing amino acids with reducing compounds (Vamin 14 and Freamine III 8.5%) was less than 10% of the added amount. Dehydroascorbic acid degraded approximately in parallel with ascorbic acid, and it contributed to the total available ascorbate activity. The addition of air to TPN mixtures in multilayered bags caused accelerated degradation of both ascorbic acid and dehydroascorbic acid. It is concluded that TPN mixtures compounded in multilayered bags can be safely assigned extended shelf lives, especially if compounded using an amino acid with reducing activity. This is principally due to the protective effect of the bag wall in preventing oxygen transmission, the cause of ascorbic acid oxidation, because oxygen transmission through the bag wall is minimized during storage. TPN mixtures stored in EVA bags should be administered within 2-4 d of compounding, depending on the amino acid infusion used.

Air

Studies on the cholesterol synthesis in the human adipose tissue. II. Mechanism of metabolic shifts and regulation of cholesterol through ascorbic acid.

Ratios of yields of cholesterol in different media with different metabolites (200 estimations) show that if the initial normal quantity in a system is X, glucose makes it 4X which is then reduced by the amino acids and ascorbic acid. Alanine, Serine, Threonine, Cysteine, Cystine and Lysine reduce it to 2X. Glycine, Valine, Leucine, Aspartic, Phenylalanine, Tyrosine, Tryptophan roughly increase it to 6X. Other amino acids are intermediate between the above sets. Balanced amino acids and fatty acids keep up the glucose value 4X but the individual amino acids behave as above. Ascorbic acid reduces the quantity of cholesterol everywhere from one third to one half bringing it back to normal X. Since it is not directly involved in the cholesterol synthesis, it must be acting through the pyridine nucleotides. The interesting feature is that the C/H ratio in the amino acids corresponds to the yield of cholesterol with respect to glucose saline yield = 1. Mechanism of metabolic shifts has been discussed on the empirical basis as also on the basis of the change in ratios of NAD+ (NADP+) to NADH2 (NADPH2) through ascorbic acid.

Acetates

Growth, ascorbic acid and iron contents of tissues of young guinea-pigs whose dams received high or low levels of dietary ascorbic acid or Fe during pregnancy and suckling.

1. Guinea-pig dams were fed on purified diets containing high (5 g/kg diet plus 1 g/l drinking water) or moderate (0.5 g/kg diet) levels of ascorbic acid, in combination with high (1 g/kg diet) or moderate (0.043 g/kg diet) levels of iron, during pregnancy and suckling. Their offsprings' diets contained 0.1 g ascorbic acid/kg and 0.04 g Fe/kg. 2. High ascorbic acid intake clearly enhanced both tissue ascorbate and Fe storage in the dams, and high Fe intake increased both the dams' and the pups' tissue Fe stores. 3. In the animals receiving high Fe intake, a co-existing high ascorbate intake by the dams reduced the growth rate of the offspring, but only during the early stages of development, not during the later stages of post-weaning growth. All the pups' tissue ascorbate levels fell after weaning, but those born of the dams receiving the high ascorbic acid diets did not fall to levels lower than those of the other pups. 4. Thus, although certain disadvantages to the offspring resulting from very-high ascorbic acid intake by pregnant guinea-pig dams were detected, these did not include permanently increased ascorbate requirements, and hence a progression to scurvy as the pups grew and matured.

Animals

Alleviating mortality associated with a vitamin E-selenium deficiency by dietary ascorbic acid.

Adding ascorbic acid to a practical ration deficient in vitamin E and selenium for the growing duck substantially reduced associated mortality. The continued appearance of various myopathies but absence of vascular faults supported implication of a reduced de novo ascorbate synthesis as part of the syndrome. Presumably, alleviation of this induced secondary inadequacy with its more lethal pathology was the primary reason for the lower death rate.

Animal Feed

Isolation and characterization of rat hepatic ascorbic acid-2-sulfatases.

Ascorbic acid-2-sulfatase was isolated from rat liver by a multistep procedure. DEAE Sephacel ion-exchange chromatography resolved crude ascorbic acid-2-sulfatase into cationic and anionic fractions. These fractions were purified 75- and 230-fold, respectively. The comparative biochemical properties suggest that arylsulfatase B is responsible for the cationic ascorbic acid-2-sulfatase activity, while arylsulfatase A appears to be responsible for the anionic ascorbic acid-2-sulfatase activity. Partially purified arylsulfatase A hydrolyzed ascorbic acid-2-sulfate at 4% the rate of p-nitrocatechol sulfate hydrolysis, while arylsulfatase B hydrolyzed ascorbic acid-2-sulfate at 0.6% the p-nitrocatechol sulfate rate.

Animals

Interactions between hepatic ascorbic acid, cytochrome P-450 and lipids in female guinea pigs with different ascorbic acid intake.

Changes in serum and liver lipids, hepatic ascorbic acid (AA) and cytochrome P-450 were investigated in female guinea pigs divided into three groups with different AA intake in drinking water (10, 100 and 1000 mg AA per liter) for 10 weeks. Serum and liver total cholesterol significantly decreased in guinea pigs receiving 100 and 1000 mg AA per liter of drinking water when compared with guinea pigs with suboptimal AA intake (10 mg/l). Similarly, serum triglycerides were decreased in the groups with higher AA intake. Liver AA concentration increased significantly in accordance with rising AA doses. High AA intake (1000 mg/l) for 10 weeks resulted in significant increase of both cytochromes P-450 and cytochrome b5 and total haeme content in liver microsomes when compared to guinea pigs with suboptimal AA intake. A significant positive correlation between hepatic AA concentration and cytochrome P-450 content was observed. A close negative correlation between liver total cholesterol and cytochrome P-450 content in hepatic microsomes was also seen. Long-term suboptimal AA intake may unfavourably alter the blood and liver lipid profile as well as the capacity of hepatic drug metabolizing enzymes in both male and female guinea pigs.

Animals

Type II epithelial cells of the lung. VII. The effect of ascorbic acid and glutathione.

Ascorbic acid and glutathione deficiencies during the isolation of tissue cells could compromise metabolic functions involving their participation. This possibility was assessed by supplementing the medium used in the isolation of rabbit pulmonary type II cells with varying concentrations and combinations of ascorbic acid, glutathione, and glutathione's constituent amino acids, glutamine, cysteine, and glycine. Isolated type II cells were monitored for changes in their biosynthetic capabilities by examination of 3H-choline and 3H-leucine incorporation into phospholipids and proteins. In the presence of glutathione's constituent amino acids and low concentrations of ascorbic acid (0.056 and 0.56 mM) incorporation of radiolabeled choline and leucine by isolated type II cells was decreased by 50% compared with unsupplemented controls. Morphologic examination revealed that type II cells isolated in the presence of glutathione's constituent amino acids had reduced surface membrane blebbing and dilation of endoplasmic reticulum cisternae compared with unsupplemented controls. Intracellular glutathione levels in these cells was also increased by 100% compared with those isolated without the constituent amino acids. Type II cells isolated in the presence of ascorbic acid and the constituent amino acids attached to culture dishes with a 5-fold greater efficiency compared with unsupplemented controls. The supplementation of type II cell's isolation media with ascorbic acid and glutathione's constituent amino acids produces several beneficial affects possibly through reducing intracellular oxidant damage, although other mechanisms of action cannot be ruled out.

Animals

Ascorbic acid and fertility.

Ascorbic acid has long been associated with fertility, but no consistent study of its mechanism of action in reproductive tissues has been made. This article considers how three of ascorbic acid's principal functions, namely its promotion of collagen synthesis, its role in hormone production, and its ability to protect cells from free radicals, may explain its reproductive actions. Data relating to both ovary and testis are reviewed since ascorbate accumulates in both tissues. Both gonads exhibit cycles of tissue remodeling and of peptide and steroid secretion that can be assumed to be ascorbate-dependent. Ascorbic acid may also prevent gametes from damage by free radicals during production and fertilization. Preliminary data on the concentrations of ascorbic acid in serum and follicular fluid from women undergoing in vitro fertilization are presented. They suggest that the supply of ascorbic acid to the ovary might be a limiting factor in the ability of the preovulatory follicle to grow in response to gonadotropin stimulation. It is concluded that ascorbic acid is a key compound in gonadal physiology on which further research is needed and that a reappraisal of its potential clinical value in the treatment of various types of male and female infertility would be timely.

Animals

Participation of urinary Na+, K+, pH, and L-ascorbic acid in the proliferative response of the bladder epithelium after the oral administration of various salts and/or ascorbic acid to rats.

Changes in urinary parameters (particularly electrolyte levels and pH), and DNA synthesis and the morphology of the bladder epithelium were investigated in rats that were fed for 4 or 8 wk on diets containing various Na, K, Mg or Ca carbonate salts, with or without L-ascorbic acid (AsA). [The carbonate salts were fed at a level of 3% in the diet, and AsA or AsA-Na was administered at 5% in the diet. NH4Cl was at 1% in the diet.] The effects of treatment with NH4Cl (used as a urine acidifier), and of combined treatment with sodium ascorbate (AsA-Na) and NH4Cl were also investigated. Urinary pH was significantly elevated in groups given NaHCO3, K2CO3, AsA + NaHCO3, AsA + K2CO3 and AsA-Na, whereas treatment with AsA or NH4Cl alone caused a significant drop in urinary pH. An increase in urinary electrolytes or ascorbic acid was associated with the corresponding dosing regimen. DNA synthesis in the bladder epithelium was increased in groups given NaHCO3, K2CO3, AsA + NaHCO3, AsA + K2CO3 or AsA-Na. Furthermore, all treatments that induced an elevation of DNA synthesis also induced some morphological alterations in the bladder epithelium. The administration of AsA in conjunction with NaHCO3 or K2CO3 induced levels of change greater than those with either salt alone. In contrast, the degree of response in the bladder epithelium of rats given AsA-Na was reduced by the simultaneous administration of NH4Cl. These results suggest that the degree of DNA synthesis and/or morphological alteration in the rat-bladder epithelium after treatment with various bases may depend on changes in urinary concentrations of Na+ or K+ ions and/or pH, and the presence of ascorbic acid in the urine. The results are discussed in relation to the possible promotion by various treatment regimens (salts +/- AsA) of urinary bladder carcinogenesis.

Administration, Oral

Resolution of the facilitated transport of dehydroascorbic acid from its intracellular accumulation as ascorbic acid.

We performed a detailed kinetic analysis of the uptake of dehydroascorbic acid by HL-60 cells under experimental conditions that enabled the differentiation of dehydroascorbic acid transport from the intracellular reduction/accumulation of ascorbic acid. Immunoblotting and immunolocalization experiments identified GLUT1 as the main glucose transporter expressed in the HL-60 cells. Kinetic analysis allowed the identification of a single functional activity involved in the transport of dehydroascorbic acid in the HL-60 cells. Transport was inhibited in a competitive manner by both 3-O-methyl-D-glucose and 2-deoxy-D-glucose. In turn, dehydroascorbic acid competitively inhibited the transport of both sugars. A second functional component identified in experiments measuring the accumulation of ascorbic acid appears to be associated with the intracellular reduction of dehydroascorbic acid to ascorbic acid and is not directly involved in the transport of dehydroascorbic acid via GLUT1. Transport of dehydroascorbic acid by HL-60 cells was independent of the presence of external Na+, whereas the intracellular accumulation of ascorbic acid was found to be a Na(+)-sensitive process. Thus, the transport of dehydroascorbic acid via glucose transporters is a Na(+)-independent process which is kinetically and biologically separable from the reduction of dehydroascorbic acid to ascorbic acid and its subsequent intracellular accumulation.

3-O-Methylglucose

Inhibition of paraquat accumulation in rabbit kidney cortex slices by ascorbic acid.

Effects of ascorbic acid and Fe2+ on the accumulation of paraquat into rabbit kidney cortical slices were studied. Ascorbic acid showed dose-dependent inhibition of paraquat accumulation. Fe2+ had a more powerful stimulatory effect on the lipid peroxidation of cortical slices than ascorbic acid, but it did not reduce paraquat accumulation. These results suggest that ascorbic acid is able to inhibit the accumulation of paraquat independent of its peroxidative action, and that ascorbic acid may serve as an antidote for the toxicity of paraquat.

Animals