Multifaceted superoxide dismutase 1 expression in amyotrophic lateral sclerosis patients:a rare occurrence?

Amyotrophic lateral sclerosis(ALS)is a neuromuscular condition resulting from the progressive degeneration of motor neurons in the cortex,brainstem,and spinal cord.While the typical clinical phenotype of ALS involves both upper and lower motor neurons,human and animal studies over the years have highlighted the potential spread to other motor and non-motor regions,expanding the phenotype of ALS.Although superoxide dismutase 1(SOD1)mutations represent a minority of ALS cases,the SOD1 gene remains a milestone in ALS research as it represents the first genetic target for personalized therapies.Despite numerous single case reports or case series exhibiting extramotor symptoms in patients with ALS mutations in SOD1(SOD1-ALS),no studies have comprehensively explored the full spectrum of extramotor neurological manifestations in this subpopulation.In this narrative review,we analyze and discuss the available literature on extrapyramidal and non-motor features during SOD1-ALS.The multifaceted expression of SOD1 could deepen our understanding of the pathogenic mechanisms,pointing towards a multidisciplinary approach for affected patients in light of new therapeutic strategies for SOD1-ALS.

Research Progress on Synthesis of Superoxide Dismutase Mimics

Research on the synthesis of superoxide dismutase mimics by chemical and biologi-cal synthetic methods were reviewed.The advantages and limitations were analyzed.A prospect for the future development of superoxide dismutase mimics is proposed.

The landscape of cognitive impairment in superoxide dismutase 1-amyotrophic lateral sclerosis

Although mutations in the superoxide dismutase 1 gene account for only a minority of total amyotrophic lateral sclerosis cases,the discovery of this gene has been crucial for amyotrophic lateral sclerosis research.Since the identification of superoxide dismutase 1 in 1993,the field of amyotrophic lateral sclerosis genetics has considerably widened,improving our understanding of the diverse pathogenic basis of amyotrophic lateral sclerosis.In this review,we focus on cognitive impairment in superoxide dismutase 1-amyotrophic lateral sclerosis patients.Literature has mostly reported that cognition remains intact in superoxide dismutase 1-amyotrophic lateral sclerosis patients,but recent reports highlight frontal lobe function frailty in patients carrying different superoxide dismutase 1-amyotrophic lateral sclerosis mutations.We thoroughly reviewed all the various mutations reported in the literature to contribute to a comprehensive database of superoxide dismutase 1-amyotrophic lateral sclerosis genotype-phenotype correlation.Such a resource could ultimately improve our mechanistic understanding of amyotrophic lateral sclerosis,enabling a more robust assessment of how the amyotrophic lateral sclerosis phenotype responds to different variants across genes,which is important for the therapeutic strategy targeting genetic mutations.Cognition in superoxide dismutase 1-amyotrophic lateral sclerosis deserves further longitudinal research since this peculiar frailty in patients with similar mutations can be conditioned by external factors,including environment and other unidentified agents including modifier genes.

Purification and Properties of Superoxide Dismutase(SOD) in Allium Sativum

Superoxide dismutases(SODs) were purified to homogeneity from Allium Sativum by means of ammonium sulfate precipitation and column chromatography with DEAE-cellulose(DE52) and Sephadex G-75. Based on sodium dodecyl sulfate\|polyacrylamide gel electrophoresis(SDS-AGE), Allium Sativum is predicted to contain four SODs. The molecular weights of the native SODs are 41 3 kD, 37 0 kD, 35 2 kD and 31 0 kD, which consist of subunits of 20 7 kD, 18 4 kD, 17 7 kD and 15 4 kD respectively. Because of their specific sensitivity to hydrogen peroxide, cyanogens potassium and chloroform\|alcohol, the SODs in Allium Sativum appear to be Cu, Zn-SOD isoenzymes. The isoelectric analysis indicates that three of the four isoenzymes are acidic proteins with isoelectric points at pH 3 5, 3 7 and 4 0, respectively, and the fourth one is a basic protein with isoeletric point at pH 8 5.

Differential Expression of Two Cytosolic Ascorbate Peroxidases and Two Superoxide Dismutase Genes in Response to Abiotic Stress in Rice

Superoxide dismutase (SOD) and ascorbate peroxidase (APX) play central roles in the pathway for scavenging reactive oxygen species in plants, thereby contributing to the tolerance against abiotic stress. Here we report the responses of cytosolic SOD (cSOD; sodCc1 and sodCc2) and cytosolic APX (cAPX; OsAPX1 and OsAPX2) genes to oxidative and abiotic stress in rice. RNA blot analyses revealed that methyl viologen treatment caused a more prominent induction of cAPXs compared with cSODs, and hydrogen peroxide treatment induced the expression of cAPXs whereas cSODs were not affected. These results suggest that cAPXs play more important roles in defense against oxidative stress compared with cSODs. It is noted that cSODs and cAPXs showed coordinate response to abscisic acid treatment which induced both sodCc1 and OsAPX2. However, cSODs and cAPXs responded differentially to drought, salt and chilling stress, which indicates that cSOD and cAPX genes are expressed differentially in response to oxidative and abiotic stress in rice.

Expression and Characterization of an Active Chimeric Protein of Human Extracellular Superoxide Dismutase and hCuZnSOD in Pichia pastoris

Mammalian cells express two isoforms of Cu- and Zn-containing superoxide dismutases（SODs）, CuZn-SOD and extracellular SOD（EC-SOD）, involved in the defense system against reactive oxygen species（ROS）. The two SODs have structurally homologous centre domain with distinct N- and C-terminuses, resulting in the different characteristics of the structure and function of the two molecules. We generated a hybrid SOD molecule（namely hy- SOD） via replacing the N- and C-terminuses of hCuZnSOD with the counterparts of hEC-SOD. The hySOD was expressed in host Pichia pastoris and the purified protein was a dimer with a molecular weight of about 34000. A series of activity analyses indicates that the hySOD is similar to hEC-SOD in heat-stability, and has the activity of protecting the host cell against heat shock and oxidative stress. Our results show evidence for the study on the compound activity of multiple SOD molecules, and may be important for understanding the relationship between structure and function of hEC-SOD and hCuZnSOD.

Characterization of a thermostable manganese-containing superoxide dismutase from inshore hot spring thermophile Thermus sp.JM1

A thermostable superoxide dismutase （SOD） from the inshore thermophile Thermus sp. JM1 was purified to homogeneity by steps of fractional ammonium sulfate precipitation, DEAE-Sepharose chromatography and Phenyl-Sepharose chromatography. The specific activity of the purified native enzyme was 1 656 U/mg. A sod gene from this strain was cloned and overexpressed in Escherichia coli （E. coli）. The prepared apo-enzyme of the purified recombinant SOD （rSOD） was reconstituted with either Fe or Mn by means of incubation with appropriate metal salts. As a result, only Mn 2＋ - reconstituted rSOD （Mn-rSOD） exhibited the specific activity of 1 598 U/mg. SOD from Thermus sp. JM1 was Mn-SOD, judging by the specific activities analysis of Fe or Mn reconstituted rSODs and the insensitivity of the native SOD to both cyanide and H 2 O 2 . Both the native SOD and Mn- rSOD were determined to be homotetramers with monomeric molecular mass of 26 kDa and 27.5 kDa, respectively. They had high thermostability at 50 ° C and 60 ° C, and showed striking stability across a wide pH span from 4.0 to 11.0.

Regulation of Content of Malondialdehyde by Siqi Decoction via Increasing Activity of Superoxide Dismutase in Blood Serum of Rats with Myocardial Ischemia

Effect of Siqi decoction on myocardial ischemia is to prevent cardiac myocyte membrane from damage associated with oxygen free radicals related to NO. To research the regulatoin of the content of malondialdehyde by Siqi decoction, an index of lipid peroxidation, via increasing activity of superoxide dismutase in blood serum of rats with Myocardial Ischemia, the model of myocardium ischemia was made in Wistar rats with posterior pituitary injection through vein in tail. Siqi decoction, Diaoxinxuekang（DK） and Fufangdanshenpian（FD）, the latter two drugs of which are effective TCM drugs of anti-myocardial ischemia at present, were administrated to the rats with myocardium ischemia for 5 days to compare the effect of them on myocardium ischemia as reference drugs via measuring the changes of the content of malondialdehyde and the activity of superoxide dismutase in the rat blood serum with myocardial ischemia. There were a remarkable increase in the activity of superoxide dismutase and a decrease in the content of malondialdehyde in the serum of the rats administered Siqi decoction compared with those of the rats in control group, p〈0.05. The contents of MDA in the serum of the prevention group rats in the experiments are lower than those of the cure group rats. Anti-Myocardium Ischemia mechanism of Siqi decoction is the regulation of the content of malondialdehyde via increasing activity of superoxide dismutase in the serum of Rats with myocardial ischemia and stimulating the activity of NOS in serum so as to increase NO concentration.

Extracellular superoxide dismutase VdSOD5 is required for virulence in Verticillium dahliae

Plants produce reactive oxygen species(ROS) to defend pathogens. To counteract this attack, certain pathogens express superoxide dismutases(SODs) to scavenge host-derived ROS. However, the roles of SODs in Verticillium dahliae, an important vascular pathogen, are not clear. Our previous study has shown that a putative extracellular SOD(VdSOD5) of V. dahliae is significantly induced by culturing in cotton tissues, suggesting that VdSOD5 may play an important role in host–pathogen interactions and virulence. Here, we showed that VdSOD5 encoded a superoxide dismutase with a cofactor copper-binding site and a functional signal peptide that can conduct protein secretion in an invertase-mutated yeast strain. The mutations in VdSOD5(ΔVdSOD5) did not change the normal vegetative growth and conidial production but reduced the virulence of V. dahliae on susceptible host cotton. Further studies showed that the transcription of Vd SOD5 was significantly up-regulated during the early stage of infection, and the loss-of-function of VdSOD5 decreased culture filtrate and fungal tissue SOD activities of V. dahliae by 74 and 28%, respectively. Compared to the wild-type strain Vd991, the ΔVdSOD5 showed the same sensitivity to the intracellular ROS generator menadione. Furthermore, nitroblue tetrazolium(NBT) staining demonstrated that VdSOD5 functioned in the detoxification of superoxides generated by host roots during infection. These results suggest that VdSOD5 of V. dahliae is an important virulence factor, secreted out of cells to combat host-derived ROS.

αB-crystallin malondialdehyde,superoxide dismutase, and lutathione peroxidase changes in X-ray irradiated rat lens

AIM: To evaluate αB-crystallin malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GPx) changes in X-ray irradiated rat lens. METHODS: Eight-week-old Sprague-Dawley male rats received X-ray irradiation to the head with rest of the body protected. The exposure dose ranged from 2 to 25 Grays (Gy). The cataract status were examined by slit lamp and rated with 'four-grade systems' post-irradiation. The lens MDA level, and the activities of SOD and GPx were measured in a short-term experiment post-irradiation, and αB-crystallin protein levels were quantified. RESULTS: The lenses of normal control and the X-ray irradiated groups with the dose up to 10 Gy remained transparent throughout the experiment. The lens first appeared tiny scatters, and even lamellar opacities in the posterior capsule 45 days post-irradiation with the dose of 15 Gy, and progressed slowly to the advance stage of cataract; while, for the higher dose (25 Gy), the opacity of lens appeared much earlier, and progressed more rapidly to mature stage of cataract within 1 month. At the end of the observation (90 days post-irradiation), almost all lenses became complete opacity with the higher dose (25 Gy). The degree of lens opacity was rated accordingly. The lens MDA level was increased, and SOD and GPx activities were decreased with a dose-dependent manner post-irradiation. The αB-crystallin protein level was decreased dose-dependently at the end point of observation. CONCLUSION: Oxidative events and αB-crystallin may play important roles in the pathogenesis of cataract in X-ray irradiated rat lens.

A Novel Isoenzyme of CuZn-superoxide Dismutase from Nicotiana tobacum

An isoenzyme of CuZnsuperoxide dismutase, denoted as CuZnSODⅢ, has been separated and purified from Nicotiana Tobacum (tobacco) leaves to apparent homogeneity. Its molecular mass is 22976.6Da. It is composed of one subunit, which is consisted of 187 amine acid residues and contains 1 copper and 0.5 zinc atom. The activation energy of the thermal denaturation process has been obtained as about 143.5kJmol-1. Meanwhile, some properties of spectra were investigated.

Purification and Characterization of Superoxide Dismutase(SOD) from Camellia Pollen

A superoxide dismutase（ SOD ） was purified to homogeneity from fresh camellia pollen by means of ammonium sulfate precipitation and column chromatography with DEAE-cellulose（ DE52 ）, Sephadex G-100 and phenyl sepharose^TM 6 Fast Flow columns. Its specific activity could reach to 4034 U/mg protein and it was determined to be Cu/ Zn-SOD according to its different sensitivities to different inhibitors. The molecular weight of the SOD and its subunit were 69500 and 34700, respectively, based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS- PAGE）, which implicates that the SOD in camellia pollen is a dimmer composed of two identical subunits. The isoelectric point of the enzyme was determined to be 4. 1 by isoelectric focusing electrophoresis and the N-terminal amino acid was identified to be Gly by the DNS-Cl method. Its α-Helix was also calculated to be approximately 21.8% according to the circular dichroism（CD） spectra.

Combined effects of temperature and copper ion concentration on the superoxide dismutase activity in Crassostrea ariakensis

Superoxide dismutase（SOD） is a crucial antioxidant enzyme playing the first defense line in antioxidant pathways against reactive oxygen species in various organisms including marine invertebrates. There exist mainly two specific forms, Cu/Zn-SOD（SOD1） and Mn-SOD（SOD2）, in eukaryotes. SODs are known to be concurrently modulated by a variety of environmental stressors. By using central composite experimental design and response surface method, the joint effects of water temperature（18–34°C） and copper ion concentration（0.1–1.5 mg/L） on the total SOD activity in the digestive gland of Crassostrea ariakensis were studied. The results showed that the linear effect of temperature was highly significant（P〈0.01）, the quadratic effect of temperature was significant（P〈0.05）; the linear effect of copper ion concentration was not significant（P〉0.05）, while the quadratic effect of copper ion concentration was highly significant（P〈0.01）; the interactive effect of temperature and copper ion concentration was not significant（P〉0.05）; the effect of temperature was greater than that of copper ion concentration. The model equation of digestive gland SOD enzyme activity towards the two factors of interest was established, with R2 and predictive R2 as high as 0.961 6 and 0.820 7, respectively, suggesting that the goodness-offit to experimental data be very satisfactory, and could be applied to prediction of digestive gland SOD activity in C. ariakensis under the conditions of the experiment. Our results would be conducive to addressing the health of aquatic animals and/or to detecting environmental problems by taking SOD as a potential bioindicator.

Synphilin-1 siRNA increases superoxide dismutase expression in a rat model of Parkinson’s disease

BACKGROUND： Synphilin-1 has been shown to be involved in the pathogenesis of Parkinson＇s disease （PD）, and superoxide dismutase （SOD） reduction might induce PD onset. To date, the precise effect of synphilin-1 on SOD expression remains poorly understood. OBJECTIVE： To explore the influence of synphilin-1 small interfering RNA （siRNA） on SOD expression in a rat model of PD. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment was performed at the Laboratory of Neurology, Affiliated Hospital of Qingdao University Medical School from June to October 2008. MATERIALS： 6-hydroxydopamine was purchased from Wuhan Boster, China. METHODS： A total of 40 male, Wistar rats were randomly assigned to PD, siRNA, siRNA negative control, and control groups, with 10 rats in each group. Rats from the PD, siRNA, and siRNA negative control groups were injected with 6-hydroxydopamine into the right substantia nigra to establish PD models. In addition, synphilin-1 siRNA and siRNA negative control sequences were separately injected into the right substantia nigra of siRNA, and siRNA negative control groups. The control group rats were treated with a mixture of ascorbic acid and normal saline. MAIN OUTCOME MEASURES： Synphilin-1 and SOD protein and mRNA expression in the substantia nigra was detected by immunohistochemistry and in situ hybridization. RESULTS： Synphilin-1 and SOD protein and mRNA expression were significantly decreased in PD and siRNA negative control groups （P 〈 0.05）. However, the siRNA group exhibited opposite effects. CONCLUSION： Synphilin-1 resulted in altered SOD expression, which suggested a protective role for synphilin-1 siRNA in an animal model of PD.

Superoxide dismutase genes in Pyropia haitanensis:molecular cloning,characterization and mRNA expression

Superoxide dismutase（SOD,EC 1.15.1.1） is the first and most important line of cellular defense against oxidative stress.In this study,based on unigene sequences of P.haitanensis,three full-length Ph SOD genes were obtained by RACE technology,and named Ph MSD,Ph CSD1 and Ph CSD2.The full-length c DNAs of these genes comprised973,1 029 and 954 nucleotides,respectively.The c DNAs encoded proteins of 224,134 and 216 amino acids,with isoelectric points of 5.75,4.65 and 10.74,respectively.Based on their conserved motifs and phylogenetic tree analysis,the three Ph SODs were divided into two SOD types：Ph MSD is a Mn-SOD,and Ph CSD1 and Ph CSD2 are Cu Zn-SODs.q PCR was used to measure the expression of the three Ph SOD genes in different life phases of P.haitanensis,and under different periods of high-temperature stress and different levels of desiccation.In the different life phases of P.haitanensis,the expression levels of the three Ph SODs were all significantly higher in the thallus than in the conchocelis.During high-temperature and desiccation stress,the expression levels of Ph CSD1 and Ph CSD2 were highly induced by the O2-content,but the expression level of Ph MSD was highly depressed by high temperature stress and showed no significant change during desiccation.

Studies on Free Radical EPR,Superoxide Dismutase Water-soluble Lipidperoxide in Tears

By means of electron paramagnetic resonance(EPR), chemistry luminescence and fluorescent spectroscopy, the free radical, superoxide dismutase and water soluble lipid peroxide in tears of normal eyes(150 eyes, 100 cases), Moorens ulcer(9 eyes), coreal grafts rejection(16 eyes) were studied. The results showed that the spin density of the free radical was 36(±058)×1012 spins/mL tear, the content of the superoxide dismutase(SOD) was 384(±145) ng/mL tear, the opposite fluorescent density of the water soluble lipid peroxide was 12912(±1691) U/mL tear in normal tears. The normal values are 25—48×1012 spins/mL tear for free radical, 239—529 ng/mL tear for SOD, 9598—16225 U/mL tear for water soluble lipid peroxide. There are significant differences in different eyes, different sexes and different ages. The free radical and lipid peroxide are higher obviously in the tears of patients with Moorens ulcer and rejected corneal grafts, compared with those of the normal control subjects(P<001), SOD is lower evidently(P<001). The above fact shows the pathogenic mechanism of Moorens ulcer and keractoplasty rejection is significantly related to toxicities injuring effect of the free radical. These results have provided important experimental data for studying lacrimalogy and new methods for clinical diagnosis and treatrment.

Screening proteins that interact with mutant superoxide dismutase 1 from familial amyotrophic lateral sclerosis using a yeast two-hybrid system

The present study screened a human fetal brain cDNA library to find the proteins that interact with mutant superoxide dismutase 1 （SOD1） using a yeast two-hybrid system. Using BLAST software, 15 real proteins which interacted with mutant SOD1 were obtained, including 8 known proteins （protein tyrosine-phosphatase non-receptor type 2, TBCl D4, protein kinase family, splicing factor, arginine/serine-rich 2, SRC protein tyrosine kinase Fyn, β-sarcoglycan; glycine receptor a2, microtubule associated protein/microtubule affinity-regulating kinase 1, ferritin H chain）, and 7 unknown proteins. Results demonstrated interaction of mutant SOD1 with microtubule associated protein/microtubule affinity-regulating kinase 1 and β-sarcoglycan.

Purification and biochemical characterization of manganesecontaining superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3

Thermostable SOD is a promising enzyme in biotechnological applications. In the present study, thermo-phileGeobacillussp.EPT3 was isolated from a deep-sea hydrothermal field in the East Pacific. A thermo-stable superoxide dismutase （SOD） from this strain was purified to homogeneity by steps of fractional am-monium sulfate precipitation, DEAE-Sepharose chromatography, and Phenyl-Sepharose chromatography. SOD was purified 13.4 fold to homogeneity with a specific activity of 3 354 U/mg and 11.1% recovery. SOD fromGeobacillussp. EPT3 was of the Mn-SOD type, judged by the insensitivity of the enzyme to both KCN and H2O2. SOD was determined to be a homodimer with monomeric molecular mass of 26.0 kDa. It had high thermostability at 50＆#176;C and 60＆#176;C. At tested conditions,SOD was relatively stable in the presence of some inhibitors and denaturants, such asβ-mercaptoethanol （β-ME）, dithiothreitol （DTT）, phenylmethylsulfonyl fluoride （PMSF）, urea, and guanidine hydrochloride.Geobacillussp. EPT3 SOD showed striking stability across a wide pH range from 5.0 to 11.0. It could withstand denaturants of extremely acidic and alkaline conditions, which makes it useful in the industrial applications.

Effects of basic fibroblast growth factor on superoxide dismutase activity and malondialdehyde content in the rat brain following intracerebral hemorrhage

BACKGROUND： Studies have confirmed that basic fibroblast growth factor （bFGF） promotes neuronal survival and neurite outgrowth. OBJECTIVE： To compare and verify the effects of bFGF on superoxide dismutase activity and malondialdehyde content in rat brain tissues surrounding a hemorrhagic lesion, as well as the hippocampus at the hemorrhagic side. DESIGN, TIME AND SETTING： The randomized, controlled, neurobiological study was performed at the Science Experimental Center and Research Laboratory, Guangxi Medical University, China, from September to December 2006. MATERIALS： Ninety-two adult, healthy, Wistar rats of equal gender were used to establish intracerebral hemorrhage by infusing type VII collagenase into the left internal capsule. Type Ⅶ collagenase （Sigma, USA）, superoxide dismutase and malondialdehyde kits （Jiancheng, China）, and bFGF （Institute of Bioengineering, Ji＇nan University, China） were used for this study. METHODS： Ninety successfully lesioned rats were equally and randomly divided into three groups. Rats in the bFGF group were intramuscularly injected daily with bFGF （8 ug/kg）. Rats in the saline control group received an equal volume of saline. The rats in the model group did not receive other interventions. Superoxide dismutase activity was measured using the xanthine oxidase method. Malondialdehyde contents were detected using the thiobarbituric acid method. MAIN OUTCOME MEASURES： At 1, 3, and 7 days following intracerebral hemorrhage, superoxide dismutase and malondialdehyde were determined in the brain tissue surrounding the hematoma and in the hippocampus in the affected hemisphere. RESULTS： In brain tissue surrounding the hematoma, superoxide dismutase activity was significantly increased in the bFGF group at 3 and 7 days after intracerebral hemorrhage compared with the saline control group, whereas malondialdehyde content was significantly decreased （P 〈 0.05）. In the hippocampus, superoxide dismutase activity was significantly increased in the bFGF group at 7 days following intracerebral hemorrhage compared with the saline control group, whereas malondialdehyde content was significantly decreased （P 〈 0.05）. At 1, 3, and 7 days after intracerebral hemorrhage, there was no significant difference between the saline control group and the model group with regards to parameter or brain region （P 〉 0.05）. CONCLUSION： Increased superoxide dismutase activity and decreased malondialdehyde content were detected in tissue surrounding the hematoma, as well as the ipsilateral hippocampus, of intracerebral hemorrhage rats treated with bFGF. Changes in these parameters were detected earlier in tissue adjacent to the lesion, compared with the ipsilateral hippocampus.

EFFECTS OF SUPEROXIDE DISMUTASE ON SOME BIOLOGICAL CHARACTERISTICS OF LUNG CANCER CELLS IN VIVO AND IN VITRO

The present study observed the effects of superoxide diamutase (SOD) and its Inhibitor, diethyldithiocarbamate (DDC), on the metastasis of Lewis Lung cancer and some biological characteristics of A548 lung cancer cell line. It was found that SOD and DDC inhibited significantly the metastasis of Lewis lung cancer In C57 BL mice, which the effect of DDC was more significant than that of SOD, and decreased the proliferation of A549 lung cancer cell and its transplantation rate in nude mice.