Effect of aspirin and vitamins C and E on synovial rheumatoid arthritic and other cells

Summary Normal and rheumatoid arthritic human synovial cells, normal rat muscle and bone cells, were cultured with combinations of aspirin (acetylsalicytic acid), vitamins C and E. Aspirin reduced percent growth of all cells by about 1/5 relative to controls. High vitamin C eradicated arthritic cells. In combinations, vitamin C was most important in eradicating arthritic cells. A low-aspirin, low-vitamin C combination was most effective in reducing arthritic cell populations, while having little effect on normal cells. Vitamin E retarded but did not prevent the action of vitamin C.Acknowledgment. This work was supported by NRC grant No. A 6445 to Dr Bhakthan, Department of Kinesiology, Simon Fraser University.     

Chemical evidence for interactions between vitamins E and C

Experimental proof is provided for interactions between radicals of vitamin E/vitamin C as generated by air-oxidized lipids (liquid fraction of subcutaneous chicken fat). Using ESR spectroscopy, hydrogen atom exchange is shown to take place between vitamin C and the radical of vitamin E. Sequential consumption of these two vitamins in oxidized lipid, first vitamin C then vitamin E, is demonstrated by means of differential pulse polarography. These results elucidate the in vitro radical scavenging functions attributed to vitamin E and vitamin C as well as their synergism in lipid antioxidation.     

Chemical evidence for interactions between vitamins E and C

Summary Experimental proof is provided for interactions between radicals of vitamin E/vitamin C as generated by air-oxidized lipids (liquid fraction of subcutaneous chicken fat). Using ESR spectroscopy, hydrogen atom exchange is shown to take place between vitamin C and the radical of vitamin E. Sequential consumption of these two vitamins in oxidized lipid, first vitamin C then vitamin E, is demonstrated by means of differential pulse polarography. These results elucidate the in vitro radical scavenging functions attributed to vitamin E and vitamin C as well as their synergism in lipid antioxidation.The authors very much thank Dr A. Dieffenbacher and P. Ducret for the preparation of the chicken fat fraction.     

Reproductive consequences of mega vitamin E supplements in female rats

Summary Female rats fed 0, 25, 2500 and 10,000 IU vitamin E/kg diet for 3 months were examined for reproductive performance. On 10,000 IU vitamin E/kg diet, the fertility of inseminated rats was significantly reduced as compared to rats given normal or nutritional levels of vitamin E.This work was supported by research grants from the National Research Council of Canada and the Research Committee of The University of British Columbia. We thank Mrs Virginia Green for her help in statistical analysis. Reprint requests should be addressed to Dr I. D. Desai, Professor of Nutrition.     

A highly active glutathione peroxidase in red blood cells of normal and acatalasemic mice

The activities of catalase (E.C.1.11.1.6) and glutathione peroxidase (E.C.1.11.1.9) were compared in red blood cells from humans, ducks and normal and acatalasemic mice. In the cells from both strains of mice, an equally high activity of GSH-Px was found which could be inhibited completely by iodoacetate but was not sensitive to N-ethylmaleimide.     

Antioxidant and prooxidant roles of copper in Tween 20-induced hemolysis of hamster and pig erythrocytes containing marginal vitamin E

The concentration-dependent effects of copper acting either as an antioxidant or as a prooxidant were examined in vitro using Tween 20-induced hemolysis. When cupric ion concentration was more than 10 M, free copper(II) acted as a prooxidant; both extensive hemolysis and production of unknown thiobarbituric acid-reactive substance occurred in hamster and pig erythrocytes irrespective of vitamin E status. However, when cupric ion concentration was 2–4 M in the incubation medium, copper showed a clear antioxidant activity, reducing both hemolysis and malondialdehyde production induced either by diluted peroxide-containing Tween 20 with ascorbic acid and sodium azide in vitamin E-deficient hamster erythrocytes, or by peroxide-containing Tween 20 in pig erythrocytes containing marginal amounts of vitamin E. Copper(II) is taken up by the erythrocytes, where copper(I)-complexes may contribute to the protection of cells with membrane vitamin E against oxidative radical attack.     

Vitamin C and the immune response.

The inclusion of vitamin C in the drinking water of BALB/c mice was without effect on the humoral antibody response to sheep red blood cells and bacterial lipopolysaccharide. However, there was a significantly increased cell-mediated immune response as determined by increased T-lymphocyte responses to concanavalin A. This might suggest a mechanism, along with interferon enhancement, for the possible protection by vitamin C against some viral infections.     

Vitamin D receptors in heart: effects on atrial natriuretic factor.

We report that receptors for vitamin D exist in distinct regions of the heart in female and male mice, predominantly in the right atrium where most of the cardial atrial natriuretic peptide (ANF) is produced. Tritiated 1,25-dihydroxyvitamin D3 (1,25-D3, vitamin D, soltriol) and ANF are colocalized in nuclei and cytoplasm respectively in identical cardiomyocytes. Changes of ANF tissue and blood levels under dietary deficiency and treatment with 1,25-D3 suggest direct genomic actions of vitamin D on myoendocrine cells of the atrium for the regulation of ANF manufacture and secretion. These results were obtained by combining thaw-mount autoradiography with immunocytochemistry using tritiated 1,25-D3 and an antibody against rat ANF. This antibody was also used in a radioimmunoassay to determine atrial natriuretic factor in plasma, atria and ventricles of normal or vitamin D-deficient mice.     

A highly active glutathione peroxidase in red blood cells of normal and acatalasemic mice

Summary The activities of catalase (E.C.1.11.1.6) and glutathione peroxidase (E.C.1.11.1.9) were compared in red blood cells from humans, ducks and normal and acatalasemic mice. In the cells from both strains of mice, an equally high activity of GSH-Px was found which could be inhibited completely by iodoacetate but was not sensitive to N-ethylmaleimide.Acknowledgment. This work was supported by a postdoctoral fellowship from the Gesellschaft für Strahlen- und Umwelt-forschung, München, Federal Republic of Germany, and was performed under contract with the U. S. Energy Research and Development Administration at the University of Rochester Biomedical and Environmental Research Project and has been assigned Report. No. UR-3490-1138.     

Vitamin C and the immune response

Summary The inclusion of vitamin C in the drinking water of BALB/c mice was without effect on the humoral antibody response to sheep red blood cells and bacterial lipopolysaccharide. However, there was a significantly increased cell-mediated immune response as determined by increased T-lymphocyte responses to concanavalin A. This might suggest a mechanism, along with interferon enhancement, for the possible protection by vitamin C against some viral infections.We thank Miss Wendy Treat for excellent technical assistance.     

Vitamin D receptors in heart: Effects on atrial natiuretic factor

We report that receptors for vitamin D exist in distinct regions of the heart in female and male mice, predominantly in the right atrium where most of the cardial atrial natriuretic peptide (ANF) is produced. Tritiated 1,25-dihydroxyvitamin D₃ (1,25-D₃, vitamin D, soltriol) and ANF are colocalized in nuclei and cytoplasm respectively in identical cardiomyocytes. Changes of ANF tissue and blood levels under dietary deficiency and treatment with 1,25-D₃ suggest direct genomic actions of vitamin D on myoendocrine cells of the atrium for the regulation of ANF manufacture and secretion. These results were obtained by combining thaw-mount autoradiography with immunocytochemistry using tritiated 1,25-D₃ and an antibody against rat ANF. This antibody was also used in a radioimmunoassay to determine atrial natriuretic factor in plasma, atria and ventricles of normal or vitamin D-deficient mice.     

Effects of lipid soluble antioxidants on cytotoxicity induced by photochemical products of cholesterol

Summary Lipid soluble antioxidants, butylated hydroxytoluene and vitamin E, were shown to suppress cytotoxicity induced by cholesterol-derived photoproducts in Chinese hamster embryo cells. These cholesterol-derived photoproducts were rather toxic.This study was supported by USPHS grant CA23684 awarded by the National Cancer Institute, NIH, DHEW.     

Protective effects of vitamin E and dithiothreitol against the hemolysis of rat and goat erythrocytes induced by Tween 20 with or without ascorbic acid and azide

Summary Exrtensive in vitro hemolysis of erythrocytes, induced in vitamie E-deficient rats by 0.001% Tween 20 with ascorbic acid and azide, or in goats by 2.5% Tween 20, could be counteracted by either the inclusion of vitamin E in the cells or by the in vitro addition of 0.25–0.4 mM dithiothreitol.     

A 41, XYY mouse

Summary Cytogenetic analyses of bone marrow and meiotic cells in an apparently normal male mouse clearly revealed the presence of an extra Y chromosome leading to 2n=41, XYY. During meiosis the sex chromosomes formed all possible types of combinations (XYY, XY/Y, YY/X and X/Y/Y). Compared to 40, XY normal mice, the sperm count was significantly less, but the incidence of sperm head abnormality was at the normal level.Acknowledgment. The authors are grateful to Prof. G. K. Manna, Department of Zoology, Kalyani University, for his valuable comments and Prof. M. C. Dash, School of Life Sciences, Sambalpur University for providing necessary facilities. Thankful acknowledgment to CSIR, India for a SRF awarded to RNK.     

Targeting mitochondria by α-tocopheryl succinate kills neuroblastoma cells irrespective of MycN oncogene expression

Amplification of the MycN oncogene characterizes a subset of highly aggressive neuroblastomas, the most common extracranial solid tumor of childhood. However, the significance of MycN amplification for tumor cell survival is controversial, since down-regulation of MycN was found to decrease markedly neuroblastoma sensitivity towards conventional anticancer drugs, cisplatin, and doxorubicin. Here, we show that a redox-silent analogue of vitamin E, α-tocopheryl succinate (α-TOS), which triggers apoptotic cell death via targeting mitochondria, can kill tumor cells irrespective of their MycN expression level. In cells overexpressing MycN, as well as cells in which MycN was switched off, α-TOS stimulated rapid entry of Ca(2+) into the cytosol, compromised Ca(2+) buffering capacity of the mitochondria and sensitized them towards mitochondrial permeability transition and subsequent apoptotic cell death. Prevention of mitochondrial Ca(2+) accumulation or chelation of cytosolic Ca(2+) rescued the cells. Thus, targeting mitochondria might be advantageous for the elimination of tumor cells with otherwise dormant apoptotic pathways.     

Binding of fluoresceinated lectins to normal and dinitrofluorobenzene treated human leucocytes

Summary Using fluoresceinated lectins we have shown the receptor distribution on normal human granulocytes and lymphocytes following tagging with 1-fluoro- 2, 4-dinitrobenzene (DNFB). DNP-tagged cells exhibited strong, smooth membrane staining and produced smaller patches dispersed uniformly over the entire cell surface.This work was supported by Grant CA 16503 from the National Cancer Institute.Acknowledgment. We thank Miss C. Pickett for technical help and Mrs E. Olson and N. Shear for secretarial assistance.     

Reduction in dietary vitamin E prevents onset of hypertension in developing spontaneously hypertensive rats

Summary Reduction in dietary vitamin E intake in developing spontaneously hypertensive rats abolished the onset of hypertension which is normally evident by 3 months of age.Supported by a grant No. 4-6 to C.P.-A. from the Canadian Ontario Heart Foundation.     

Wirkungen von Gallensäure und Vitamin D auf die Kalziumresorption bei der Ratte

Summary Comparison of the effects of vitamin D and the bile salt Taurochenodeoxycholate on calcium absorption in young rats suggests that bile salt enhancement of calcium absorption occurs mainly in the ileum in normal animals but occurs in both duodenum and ileum in rachitic rats. Also suggested is that bile salts mainly assist the entry of calcium into mucosal cells from the gut lumen while vitamin D chiefly aids the egress of calcium from the cells into the extracellular fluid.     

Vitamin E,catalase, manganous or cobaltous ions and dithiothreitol protect against Tween 20-induced hemolysis of vitamin E-deficient chick and kid erythrocytes

Summary The effective Tween 20 concentration at which 70% hemolysis was achieved in vitro correlated with the plasma vitamin E content of chicks (r=0.85). Addition of catalase, MnCl₂, CoCl₂ or dithiothreitol in vitro showed significant protection against the hemolysis induced by Tween 20 in vitamin E-deficient chick and kid erythrocytes.     

Binding of D-alpha-tocopherol to rat liver nuclear components.

When D-alpha-tocopherol is administered i.v. to vitamin E deficient rats, significant amounts of this vitamin are bound to a nucleoprotein complex in hepatic nuclei, and this complex can be solubilized by high concentrations of sodium chloride (0.6 M). The bound vitamin in this complex, extractable by ethanol, was found to be identical with authentic alpha-tocopherol by thin layer chromatography.