Superoxide Dismutase Deficiency Research Articles

Characterization of the Bacillus stearothermophilus manganese superoxide dismutase gene and its ability to complement copper/zinc superoxide dismutase deficiency in Saccharomyces cerevisiae

Recombinant clones containing the manganese superoxide dismutase (MnSOD) gene of Bacillus stearothermophilus were isolated with an oligonucleotide probe designed to match a part of the previously determined amino acid sequence. Complementation analyses, performed by introducing each plasmid into a superoxide dismutase-deficient mutant of Escherichia coli, allowed us to define the region of DNA which encodes the MnSOD structural gene and to identify a promoter region immediately upstream from the gene. These data were subsequently confirmed by DNA sequencing. Since MnSOD is normally restricted to the mitochondria in eucaryotes, we were interested (i) in determining whether B. stearothermophilus MnSOD could function in eucaryotic cytosol and (ii) in determining whether MnSOD could replace the structurally unrelated copper/zinc superoxide dismutase (Cu/ZnSOD) which is normally found there. To test this, the sequence encoding bacterial MnSOD was cloned into a yeast expression vector and subsequently introduced into a Cu/ZnSOD-deficient mutant of the yeast Saccharomyces cerevisiae. Functional expression of the protein was demonstrated, and complementation tests revealed that the protein was able to provide tolerance at wild-type levels to conditions which are normally restrictive for this mutant. Thus, in spite of the evolutionary unrelatedness of these two enzymes, Cu/ZnSOD can be functionally replaced by MnSOD in yeast cytosol.

Superoxide dismutase deficiency and the toxicity of the products of autooxidation of polyunsaturated fatty acids in yeast

Mutants of Saccharomyces cerevisiae, deficient in cytosolic superoxide dismutase and catalase activities were used to study the role of various oxygen species in the process of lipid peroxidation in yeast cells. Lipid peroxidation does not occur normally in yeast, because this organism is unable to form fatty acids with more than one double bond, whereas under physiological conditions, only fatty acids with at least two double bonds undergo this process. The fatty acid content of cellular lipids was modified by growing the cells in anoxia in the presence of oleic or linolenic acid. Toxic effects of oxygen were observed almost exclusively in those cells of yeast mutants deficient in superoxide dismutase, which contain linolenic acid in cellular lipids. Hypersensitivity of the mutant cells, however, results mainly from toxic effects of the products of autooxidation of extracellular fatty acids. These facts suggest that superoxide dismutases are in some way involved in preventing toxic effects of the products of lipid peroxidation and to some extent prevent the process of lipid peroxidation.

ACRODERMATITIS ASSOCIATED WITH ZINC DEFICIENCY: FEATURES AND POSTULATED MECHANISM

SummaryIt is currently unexplained why zinc deficiency should result in a cutaneous eruption of such a distinctive and characteristic distribution (acrodermatitis), affecting the groin, periorificial surfaces and periungual areas of fingers and toes. In this study, we provide ultrastructural evidence suggestive of excessive damage of the epidermis by neutrophils, with other features resembling tissue damage from free radicals. We propose that these changes are consistent with a deficiency of superoxide dismutase, a zinc metalloenzyme, which is mainly responsible for neutralizing the destructive effects of the reactive oxygen species released by neutrophils. The various zymosans on many yeast cells, including Candida, activate complement, resulting in the generation of C5a, a potent chemattractant for neutrophils, and the location of the eruption in the characteristic sites in zinc deficiency may be explained by the natural occurrence of Candida albicans in these areas.

Bovine ocular squamous cell carcinoma: superoxide dismutase and catalase levels

Considerable evidence exists for the production of free radicals in the eye region after exposure to ultraviolet light. Failure to remove these radicals adequately may be important in the predisposition of Hereford cattle to ocular carcinoma. The levels of two enzymes, superoxide dismutase and catalase, involved in the removal of superoxide anion and hydrogen peroxide, were compared in Hereford cattle predisposed to ocular carcinoma and a resistant breed, Droughtmaster cattle. The results demonstrate comparable levels of superoxide dismutase in the two breeds in both eye tissue and erythrocytes. Catalase levels also were the same in both breeds. Increased incidence of ocular carcinoma in Herefords does not appear to be due to a deficiency in either superoxide dismutase or catalase.

A modelin vitrosystem for assessing the effects of oxidant stressor agents on red cells with chemically‐induced superoxide dismutase deficiency

Abstract An in vitro test system was established to determine the oxidant Stressor effects of various agents on erythrocytes having low Superoxide dismutase (SOD) activity. The compound, diethyldithiocarbamate (DDC) was used to inhibit SOD activity in normal human erythrocytes. SOD activity was reduced to 67%, 50% and 41% of normal via the action of 1 mM, 2 mM and 4mM DDC, respectively. Five environmental agents, carbaryl, copper, nitrite, paraquat and p‐chloroaniline, were evaluated. P‐chloroaniline (5 mM) and carbaryl (2 mM) caused significantly raised methemoglobin levels (p = .001) in erythrocytes with 41% of normal SOD activity. Linear regression analyses indicated that a significant linear increase in methemoglobin levels (p = .001) occurred for erythrocytes, having reduced SOD activity, incubated with p‐chloroaniline and carbaryl.

Deficiency of erythrocyte superoxide dismutase and catalase activities in patients with malignant lymphoma and acute myeloid leukemia

The activity of erythrocyte superoxide dismutase (SOD) and catalase was determined in 38 patients with lung cancers ( n = 15), malignant lymphoma ( n = 11) and acute myeloid leukemia ( n = 12). The patients with malignant lymphoma and acute myeloid leukemia showed a significant decrease in both enzyme activities ( P < 0.01) while the patients with lung cancer (9 squamous cell carcinomas and 6 small cell carcinomas) showed normal values of SOD and catalase activities. Furthermore, the patients with malignant lymphoma and acute myeloid leukemia, in remission, but not treated with anticancer drugs, also showed a significant decrease in SOD and catalase activity. These observations therefore indicate that a deficiency of erythrocyte SOD and catalase activities is caused by cancer and varies with the type of the cancer.

Manganese-containing superoxide-dismutase deficiency in polymorphonuclear leukocytes of adults with rheumatoid arthritis.

Superoxide dismutase (SOD) is known to regulate the level of superoxide radicals inside cells. The purpose of this work was to investigate the role of SOD activity in tissue damage produced by superoxide radicals. SOD was measured in polymorphonuclear cells of patients with rheumatoid arthritis and controls. The distinct SOD activities, including manganese-containing and copper-zinc-containing enzymes, were evaluated in cytoplasma and mitochondria of human granulocytes. Except for the comparison between total SOD and cytoplasmic copper-zinc SOD, no correlation was found among the different SOD levels. Moreover, a significant decrease was observed only for cytoplasmic manganese-containing enzyme in granulocytes of adults with rheumatoid arthritis. These data confirm the necessity of evaluation of various SOD classes and suggest the interest of biochemical tests in granulocytes for early diagnosis and better comprehension of tissue damage due to inflammation.

Superoxide dismutase, glutathione peroxidase and catalase in oxidative hemolysis. A study of Fanconi's anemia erythrocytes

Superoxide dismutase, glutathione peroxidase and catalase were assayed in the erythrocytes of three patients of Fanconi's anemia. Superoxide dismutase was found to be significantly decreased, as previously reported. The enzymes metabolizing H 2O 2 are normal (glutathione peroxidase in the higher limits of the normal value). The abnormal erythrocytes were found to be as resistant (perhaps more resistant) as normal red blood cells to oxidative hemolysis induced by drugs. Malonyl dialdehyde production was found to be comparable to that of normal erythrocytes. It is concluded that a significant (30–40%) deficiency of superoxide dismutase, when associated to normal values of H 2O 2-removing enzymes, does not affect the antioxidative defense capability of erythrocytes, even in conditions of augmented oxidative injury.

A hypothesis for the molecular mechanism of tumor killing by porphyrins and light

Tumor cells have an absence or deficiency of mitochondrial superoxide dismutase. Superoxide is produced when hematoporphyrin derivative is exposed to visible light. A significant portion of the antitumor effect of porphyrin: light therapy is hypothesized to be mediated by superoxide.

Superoxide dismutase deficiency in patients with nitrogen mustard therapy-induced intravascular hemolysis

Superoxide dismutase deficiency in patients with nitrogen mustard therapy-induced intravascular hemolysis

The effect of superoxide dismutase, catalase and l-cysteine on spontaneous and on mitomycin C induced chromosomal breakage in Fanconi's anemia and normal fibroblasts as measured by the micronucleus method

Fanconi's anemia (FA) is characterized by high frequencies of chromosome aberrations and predisposition to cancer. There are two suggestions concerning the nature of the biochemical defect in FA: (a) that the condition is the result of a defect in DNA repair specifically of DNA crosslinks and (b) that the condition is associated with a deficiency in superoxide dismutase. Since it is not evident how either of these biochemical alterations would lead to the other, these hypotheses may be regarded as mutually exclusive. If a deficiency in superoxide dismutase is the primary biochemical defect in FA, then treatment with this enzyme should have a much greater effect in Fanconi than in normal cells. Furthermore, superoxide dismutase should be more effective than catalase and l-cysteine. On the repair-deficiency hypothesis, any effect of superoxide dismustase should be proportionately the same in Fanconi and normal cells, and need not be more effective than the catalase or l-cysteine. To test these predictions we have measured chromosomal breakage in 3 Fanconi fibroblast strains and 1 normal strain treated with superoxide dismutase, catalase, and l-cysteine. All 3 of the Fanconi strains were more sensitive to a DNA crosslinking agent, mitomycin C (MMC), than the normal strain and all had a high spontaneous level of chromosomal damage. Treatment by superoxide dismutase reduced both the spontaneous and MMC induced chromosomal breakage in all strains. There was no systematic difference among the Fanconi and the normal strains in the extent of the reductions. Treatment by catalase was even more effective than treatment by superoxide dismutase. We conclude that the effect of superoxide dismustase and of catalase and l-cysteine is not specific, at least in Fanconi cell strains that are MMC-sensitive. These results are in accord with the repair-deficiency hypothesis, but not with the superoxide dismutase deficiency hypothesis.

Erythrocyte superoxide dismutase deficiency in Fanconi's anaemia established by two independent methods of assay

Erythrocyte superoxide dismutase deficiency in Fanconi's anaemia established by two independent methods of assay

Erythrocyte superoxide dismutase deficiency in Fanconi's anaemia established by two independent methods of assay

Erythrocyte superoxide dismutase (SOD; E.C.1.15.1.1) levels were quantitated in human haemolysates, both by an immunological technique and by an activity assay. The erythrocyte SOD activity level in a group of seven Fanconi's anaemia patients was significantly decreased (27%, on the average) compared to a control group of nineteen healthy individuals. The fact that both the activity per antigenic unit and the electrophoretic mobility of the erythrocyte SOD was normal indicates that the deficiency in these Fanconi's anaemia patients is most probably not due to a mutation in the structural gene for the enzyme but more likely due to a disturbed mechanism regulating the SOD level in the erythrocyte.

Superoxide dismutase in bovine fetal ductus arteriosus, throacic aorta, and pulmonary and umbilical arteries.

SummarySoon after birth the lumens of the ductus arteriosus (DA) and umbilical artery (UA) are obliterated. It has been suggested that oxygen toxicity, resulting from an increased oxygen tension, is the cause of this destruction with superoxide radicals and hydroxyl radicals being implicated as mediators. A deficiency of superoxide dismutase (SOD) in these tissues was hypothesized as being responsible for an increase in the levels of superoxide and hydroxyl radicals. SOD levels were determined enzymatically and immunochemically in four tissues obtained from thirteen bovine fetuses. SOD levels in the DA and UA were found by both assays to be statistically significantly lower than that found in such permanent vessels as the pulmonary artery and thoracic aorta. These data are in support of the hypothesis that a lower level of SOD in the ductus arteriosus and umbilical artery may contribute to the rapid deterioration of these tissues upon exposure to greatly increased oxygen tensions.The kind assistance of Io...

The organelle pathology and demonstration of mitochondrial superoxide dismutase deficiency in two patients with Dubin--Johnson--Sprinz syndrome.

1. Liver biopsies from two patients with the Dubin-Johnson-Sprinz syndrome were subjected to analytical subcellular fractionation by sucrose-density-gradient centrifugation and enzymic microassays. 2. A selective deficiency of mitochondrial superoxide dismutase has been demonstrated in these patients. 3. The significance of this enzyme deficiency is discussed in relation to the organelle pathology of the syndrome.

NEWBORN RED BLOOD CELLS (RBC's), THE NATURE OF OXIDANT INJURY

Unlike adult cells, newborn RBC's readily undergo oxidant damage. The nature of the defect causing this is unclear. In order to determine those factors related to newborn RBC oxidant susceptability, cord blood was obtained from 88 term infants. As expected, acetylphenylhydrazine (APH) induced Heinz Body (HB) formation was greater in newborn vs adult RBC's (3.24 HB/RBC vs 0.27). Although newborn RBC superoxide dismutase (SOD) activity, at 91% of adult levels, was slightly but significantly lower than adult RBC's (paired T-4.0 p<0.001), higher levels of SOD were associated with greater APH induced HB and H2O2 formation (as determined by the aminotriazole - catalase inhibition assay). The addition of SOD to newborn RBC's produced additional significant increases in HB and H2O2. No correlation existed between %HbF, amount of HbF oxidized and HB or H2O2 formed although H2O2 production was 82% > in newborn RBC's. With equivalent H2O2 production, HB formed readily in newborn cells only. Glutathione peroxidase (GSH Px) levels were equivalent in newborn and adult RBC's. Despite the H2O2 formed, the fall in newborn RBC reduced glutathione (GSH) levels induced by menadione failed to exceed that of adult RBC's when incubated in the absence of glucose.These data demonstrate that SOD deficiency is not a cause of the vulnerability of newborn RBC's to oxidant damage; in fact > SOD is associated with > oxidant effect. It appears that > H2O2 generation and ineffective detoxification by GSH (inspite of “normal” levels of GSH Px) are the major determinants of oxidative damage.

NEWBORN RED BLOOD CELLS (RBC's), THE NATURE OF OXIDANT INJURY

Unlike adult cells, newborn RBC's readily undergo oxidant damage. The nature of the defect causing this is unclear. In order to determine those factors related to newborn RBC oxidant susceptability, cord blood was obtained from 88 term infants. As expected, acetylphenylhydrazine (APH) induced Heinz Body (HB) formation was greater in newborn vs adult RBC's (3.24 HB/RBC vs 0.27). Although newborn RBC superoxide dismutase (SOD) activity, at 91% of adult levels, was slightly but significantly lower than adult RBC's (paired T-4.0 p in newborn RBC's. With equivalent H2O2 production, HB formed readily in newborn cells only. Glutathione peroxidase (GSH Px) levels were equivalent in newborn and adult RBC's. Despite the H2O2 formed, the fall in newborn RBC reduced glutathione (GSH) levels induced by menadione failed to exceed that of adult RBC's when incubated in the absence of glucose. These data demonstrate that SOD deficiency is not a cause of the vulnerability of newborn RBC's to oxidant damage; in fact > SOD is associated with > oxidant effect. It appears that > H2O2 generation and ineffective detoxification by GSH (inspite of “normal” levels of GSH Px) are the major determinants of oxidative damage.

RED BLOOD CELL (RBC) ABNORMALITIES IN CHRONIC GRANULOMATOUS DISEASE (CGD) WITH MCLEOD PHENOTYPE

Unlike adult cells, newborn RBC's readily undergo oxidant damage. The nature of the defect causing this is unclear. In order to determine those factors related to newborn RBC oxidant susceptability, cord blood was obtained from 88 term infants. As expected, acetylphenylhydrazine (APH) induced Heinz Body (HB) formation was greater in newborn vs adult RBC's (3.24 HB/RBC vs 0.27). Although newborn RBC superoxide dismutase (SOD) activity, at 91% of adult levels, was slightly but significantly lower than adult RBC's (paired T-4.0 p<0.001), higher levels of SOD were associated with greater APH induced HB and H2O2 formation (as determined by the aminotriazole - catalase inhibition assay). The addition of SOD to newborn RBC's produced additional significant increases in HB and H2O2. No correlation existed between %HbF, amount of HbF oxidized and HB or H2O2 formed although H2O2 production was 82% > in newborn RBC's. With equivalent H2O2 production, HB formed readily in newborn cells only. Glutathione peroxidase (GSH Px) levels were equivalent in newborn and adult RBC's. Despite the H2O2 formed, the fall in newborn RBC reduced glutathione (GSH) levels induced by menadione failed to exceed that of adult RBC's when incubated in the absence of glucose. These data demonstrate that SOD deficiency is not a cause of the vulnerability of newborn RBC's to oxidant damage; in fact > SOD is associated with > oxidant effect. It appears that > H2O2 generation and ineffective detoxification by GSH (inspite of “normal” levels of GSH Px) are the major determinants of oxidative damage.