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.

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.

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.

Relationship of Uric Acid with Superoxide Dismutase (Sod) in Induced Hyperuricemic Rat Model

Increase uric acid levels have been found in oxidative stress. Urate radicals do not react with oxygen to form another peroxy radical, thus increasing the efficacy of uric acid as an antioxidant. Therefore, this study is designed to measure the level of uric acids and find out the relationship of uric acid with superoxide dismutase in induced hyperuricemic model. Forty male albino rats with an average weight of 180 ± 2 g were selected. The rats were grouped. The animals were fed on standard diet and given tap water ad libitum until treatment. Albino rats were divided into four groups. Group A(10)-control given only standard diet, group B(10) fed on 60% fructose with standard diet , group C(10) fed on fructose, standard diet and intraperitonially oxonic acid 250 mg/kg and group D (10) only on injection intraperotonially oxonic acid 250 mg/kg. At the end of study 10 mL of blood was drawn from heart of rats. Then blood was estimated for superoxide dismutase and uric acids done by kit methods randox-manual/Rx monza UA230/UA 233. Results: In Group C superoxide dismutase was found to be 32 % (244 mg/dL ± 2.23) more than control. In the same group the uric acid concentration was highly significantly correlated with control. Conclusion: The uric acid concentration increases when we take fructose up to 60% in our diet. It also increases superoxide dismutase concentration. More than this value may have inverse effect on the uric acid level and its role as an antioxidant may become inversed.

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.

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.

真空预冷压强对鲜枸杞SOD酶活性的影响及分子动力学模拟

为探究真空预冷处理对鲜枸杞超氧化物歧化酶(superoxide dismutase,SOD)活性的影响及其分子水平机理,本研究将宁夏新鲜枸杞置于真空预冷900、1100 Pa压强下处理,提取粗酶液进行SOD酶活性测定,并进行150 ns分子动力学模拟研究SOD酶在不同真空预冷压强处理下的内部变化,分析均方根误差、回旋半径、总能量、势能、动能、键角能、均方根涨落、氢键数量、表面结构、溶剂可及表面积的改变,进而筛选出真空预冷处理的最佳压强条件。结果显示,900 Pa处理下SOD酶活最高。分子动力学模拟结果表明,体系总能量、势能、动能及键角能均处于平衡稳定状态,压强增大使得蛋白质表面结构改变,氢键数量减少,溶剂可及表面积不断减小。在900 Pa处理下,蛋白质结构最紧密,活性口袋变宽且易与蛋白结合。而1100 Pa处理下酶蛋白活性降低,蛋白结构破坏,活性口袋变小。本研究可为研究鲜枸杞保鲜及加工提供科学参考。

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.

金匮肾气丸联合维生素D钙片治疗2型糖尿病性骨质疏松症的疗效及对血清骨转换标志物、IL-1β、MMP-9、SOD水平的影响

目的探讨金匮肾气丸联合维生素D钙片治疗2型糖尿病性骨质疏松症(T2DOP)的临床疗效,以及对患者血清骨转换标志物、白细胞介素-1β(IL-1β)、基质金属蛋白酶-9(MMP-9)、超氧化物歧化酶(SOD)水平的影响。方法选择该院2021年1月至2022年10月收治的84例T2DOP患者作为研究对象,按随机数字表法将患者分为观察组(42例)和对照组(42例)。观察组采用金匮肾气丸联合维生素D钙片治疗,对照组单用维生素D钙片治疗,治疗24周后比较两组临床疗效。治疗前后检测两组患者第1~4腰椎(L 1~4)、股骨颈的骨密度(BMD),并对两组患者进行疼痛视觉模拟量表(VAS)评分。治疗前后检测两组患者的糖代谢指标[空腹血糖(FPG)、餐后2 h血糖(2 h PG)、糖化血红蛋白(HbA1c)]、血清骨转换标志物{25羟基维生素D[25(OH)D]、β-胶原特殊序列(β-Crosslaps)、总Ⅰ型胶原氨基端延长肽(Total-P1NP)、骨钙素N端中分子片段(N-MID)}以及IL-1β、MMP-9、SOD水平。统计两组不良反应发生情况。结果观察组总有效率为95.24%,明显高于对照组的76.19%(P<0.05)。治疗后,两组L 1~4 BMD以及血清25(OH)D、Total-P1NP、N-MID、SOD水平均高于治疗前(P<0.05),VAS评分、FPG、2 h PG、HbA1c以及血清β-Crosslaps、IL-1β、MMP-9水平均低于治疗前(P<0.05)。治疗后,观察组L 1~4 BMD以及血清25(OH)D、Total-P1NP、N-MID、SOD水平均高于对照组(P<0.05),VAS评分、FPG、2 h PG、HbA1c以及血清β-Crosslaps、IL-1β、MMP-9水平均低于对照组(P<0.05)。两组不良反应总发生率比较,差异无统计学意义(P>0.05)。结论金匮肾气丸联合维生素D钙片治疗T2DOP能有效调节患者血清骨转换标志物、IL-1β、MMP-9、SOD水平,改善糖代谢与骨代谢,提高临床疗效。

Superoxide Dismutase: Therapeutic Targets in SOD Related Pathology

There are growing evidences on the role of adaptive mechanisms of all cell types in pathological processes: atherosclerosis, ischemic attack, bacterial infections, etc. All kinds of these processes involve as main mechanism oxidative stress. Aerobic organisms use oxygen in processes that accidentally or deliberately generate aggressive species for the biologic components in the form of radicals. Radicals were looked initially as “harmful” molecules and this is true for large quantities but in small or even moderate amounts these molecules prove to have a physiological role. Reactive species are highly reactive and as a consequence are short living species. Their impact is supposed to be limited in the proximity area of their formation. Instead recent evidences indicate their implications in cellular signaling suggesting that individual chemical properties of reactive species make a difference in their biological role. This paper presents superoxide, nitric oxide and peroxide radical generation under cellular changing conditions, the adapting behavior of the enzymes that synthesize and remove them as well as some therapeutic target in superoxide related pathology.

GroESL protects superoxide dismutase (SOD)— Deficient cells against oxidative stress and is a chaperone for SOD

Superoxide dismutase (SOD)-deficient Escherichia coli OX326Acells are protected against chemically-induced oxidative stress by expression of the chaperonin GroESL. This protection is equivalent to expression of superoxide dismutase even though GroESL has no inherent SOD activity. Co-overexpression of GroESL and SOD in the same cells results in higher protein yields of SOD and greater metallation of SOD when compared with expression of SOD alone. Greater metallation results in the higher specific activity of SOD that is observed in heat shock, and is not due to increased synthesis of SOD mRNA or protein.

Effect of N+ Beam Exposure on Superoxide Dismutase and Catalase Activities and Induction of Mn-SOD in Deinococcus Radiodurans

Though bacteria of the radiation-resistant Deinococcus radiodurans have a high resistance to the lethal and mutagenic effects of many DNA-damaging agents, the mechanisms involved in the response of these bacteria to oxidative stress are poorly understood. In this report, the superoxide dismutase (SOD) and catalase (CAT) activities produced by these bacteria were measured, and the change of SOD and CAT activities by 20 keV N+ beam exposure was examined. Their activities were increased by N+ beam exposure from 8x 1014 ions/cm2 to 6x1015 ions/cm2.The treatment of H2O2 and [ CHC13 +CH3 CH2OH ] and the measurement of absorption spectrum showed that the increase in SOD activity was resulted from inducible activities of MnSOD in D. radiodurans AS1.633 by N+ beam exposure . These results suggested that this bacteria possess inducible defense mechanisms against the deleterious effects of oxidization.

Binding studies of trans-resveratrol with superoxide dismutase (SOD1): Docking assessment and Thermoanalysis

The binding pursuits of trans-resveratrol(t-RSV),an amazing health supplement are investigated with an antioxidant enzyme,superoxide dismutase(SOD1).The aim of the study is to dock t-RSV on the adrenaline binding site on SOD1 in order to explore its potential to act as a safety net against amyotrophic lateral sclerosis(ALS),a fatal neurodegenerative disorder that affects motor neurons.In silico GLIDE docking methodology and in vitro microcalorimetry technique is utilized for the investigation of binding parameters of t-RSV with SOD1.The study provides useful and distinct information about the amino acids involved in the interactions at molecular level along with the nature of forces involved in binding of t-RSV with SOD1.The docking analysis using the scoring functions of Schrodinger’s Glide package depicts that GLU100,PRO28,LYS23,TRP32 residues of the peptide backbone on SOD1 interact with phenolic groups of t-RSV.The information on thermodynamic parameters,i.e.binding constant(Kb),free energy(△G)and enthalpy(△H)generated through calorimetric titrations suggests that the reaction between t-RSV and SOD1 is spontaneous and exothermic.Both the studies are found to be in close agreement with each other based as far as the magnitude of binding constant(Kb=9.9×10^4)is concerned.

Effects of Hg^(2+) on Isozymes of Peroxidase,Catalase and Superoxide Dismutase in Wheat Seedlings

[Objective] This work was aimed to explore the mechanism of Hg2+ toxicity on plants.[Method]Activities of peroxidase(POD),catalase(CAT)and superoxide dismutase(SOD)were investigated in wheat(Triticum aestivum L.)seedlings under Hg2+ stress at different concentrations.[Result]① There were no obvious effects on the growth of seedlings when the concentration of Hg2+ was lower than 0.10 mmol/L.However,toxic effects on the growth of seedling were observed when the concentration of Hg2+ was higher than 0.10 mmol/L.② Different tissues showed different resistant ability in response to Hg2+ stress.The leaves and roots of wheat seedlings were more insensitive to Hg2+ toxicity.③ CAT was more sensitive to Hg2+ stress compared to POD and SOD.[Conclusion]The toxic effect was related to the concentration of Hg2+(0.10 mmol/L).The higher concentration of Hg2+ could affect the expression of POD,CAT,and SOD isozymes in the leaves,roots of wheat seedlings and germinated seeds,which further affect the normal metabolism of membrane lipid and inhibit the growth of wheat seedlings at last.

Comparison of Superoxide Dismutase Isoenzymes in Three Shrimps

[Objective] The aim of the study is to compare the activities of superoxide dismutase(SOD) isoenzymes in three species of shrimps Penaeus japonicus, Procambarus clarkia and Litopenaeus vannamei. [Method] The experimental materials were used to measure SOD activities after pretreatment, meanwhile the differences in SOD isoenzymes from different materials were assayed by polyacrylamide gel electrophoresis (PAGE). [Result] There are specific and histological differences in SOD activities of shrimps. With a similar electrophoresis pattern and migration rate, Penaeus japonicus and Litopenaeus vannamei showed remarkable differences with that of Procambarus clarkia. [Conclusion] The result showed the differences of cognation and origin of three shrimps.

Analysis of Amylase and Superoxide Dismutase Isozymes During the Germination Process of Emmenopterys henryi Oliv Seeds

[ Objective] The study was to understand the changes of amylase（AMY） and superoxide dismutase（SOD） isozymes during the ger- mination process of Emmenopterys henryi Oliv seeds. [ Metbod] By employing polyacrylamide gel electrophoresis method, the expressions of AMY and SOD isozymes during seed germination process were analyzed. ~ Result] The main AMY bands remained strong during the whole peri- od and a new band A2 appeared in the middle and late period of seed germination. Some new SOD bands occurred at the early stage, then be- came weak or disappeared in the middle period, and band S6 became intense in the late peried. [ Conclusion.] The expression of AMY and SOD isozyme gene has temporal difference during germination of E. henryi Oliv seeds.

牛血超氧化物歧化酶(bovine superoxide dismutase)生产工艺研究

将红细胞连续分离、热变性以及超滤技术应用在牛血 SOD生产制备中 ,使 SOD生产成本大幅降低 ,实现牛血 SOD生产工业化 .实验表明 ,冷冻血球和新鲜血球在酶的收率、纯度以及比活等方面无明显差别 ;热变性、超滤、丙酮沉淀等各工艺步骤酶的活性以及收率基本稳定 ;通过重组 SOD技术使失去铜锌离子失活的酶蛋白恢复酶的催化活性 .

Combined Extracting Process and Activity Detection of Porcine Blood Superoxide Dismutase

[Objective] TO study the combined extracting process of porcine blood superoxide dismutase （SOD） and other bioactive substances and thus to provide technical basis for making full use of blood resources and large-scale production of SOD. [Method] Fibronectin, immunoglobulin, and hemoglobin were isolated from porcine blood, and SOD was extracted. Trace pyrogallol self-oxidation method to determine SOD activity was modified by optimizing the volume of pyrogallol and SOD samples, reaction temperature, and buffer pH. The specific activity of SOD was determined with the optimized extraction conditions. [ Result] The improved experimental conditions of SOD activity detection were as follows： 7 pyrogallol （50 mmol/L）, 3 ml Tris-HCI （50 mmol/L, pH 8.2）, reactive temperature at 25（3, and 10 pl SOD sample solution. The specific activity of extracted SOD was 5 056 U/mg protein. [ Conclusion] Four kinds of bioactive substance can be isolated from porcine blood by modern biological engi- neering integration technology, and the extracted SOD has better activity.

棉花SOD基因家族生物信息学分析

研究基于镉胁迫下棉花幼苗根系的转录组测序数据,鉴定获得16个棉花SOD基因,并对其进行生物信息学分析。结果表明:16个棉花SOD基因编码的蛋白长度为127~333 aa,相对分子质量为13048.69~35974.31 Da,等电点为4.43~8.50,均为非分泌蛋白,且都无跨膜结构特征,表现为亲水性;其中75%属于酸性蛋白,69%属于稳定蛋白;大部分亚细胞定位在叶绿体;16个棉花SOD可分为Fe-SOD、Mn-SOD和Cu/Zn-SOD 3个亚家族,3个亚家族均含有各自的保守基序,其中Fe-SOD和Mn-SOD亚家族亲缘关系较近,推测两者功能更相似。棉花SOD家族基因的鉴定、分析对棉花SOD基因功能及镉响应机制研究具有借鉴意义,可为筛选耐镉棉花品种提供基因资源。

CaR调节TRPC6介导的自噬对软骨细胞SOD及NOX2的影响

目的观察钙敏感受体(CaR)调节瞬时感受器电位通道6(TRPC6)介导的自噬对软骨细胞超氧化物歧化酶(SOD)、NADPH氧化酶-2(NOX2)的影响。方法2023年7月将出生3~5 d SD大鼠关节软骨用胶原酶消化后进行传代培养,取第3代培养的细胞接种于96孔板,用白细胞介素-1β(10 ng/mL)诱导软骨细胞,分为对照组、CaR(10μg/L)组、U73122(5μg/L)组和CaR(10μg/L)+U73122(5μg/L)组,共培养24 h后应用酶联免疫吸附试验检测各组上清液SOD、NOX2水平,采用实时定量-聚合酶链反应检测各组SOD、NOX2、人自噬基因Beclin-1 mRNA表达,Western-blot检测SOD、NOX2、Beclin-1蛋白表达。结果原代关节软骨细胞在显微镜下呈三角形或不规则形或多角形,分布稀疏,培养24 h后可见贴壁生长;第3代软骨细胞显示其蓝染的细胞核,细胞间质及细胞质内偶见紫红色异染颗粒;细胞核染成蓝色,细胞质区呈棕黄色。对照组软骨细胞上清液SOD水平,SOD、Beclin-1 mRNA及其蛋白相对表达量,NOX2 mRNA相对表达量均明显低于CaR(10μg/L)组,NOX2水平、NOX2蛋白相对表达量均明显高于CaR(10μg/L)组,差异均有统计学意义(P<0.05);U73122(5μg/L)组和CaR(10μg/L)+U73122(5μg/L)组软骨细胞上清液SOD水平,SOD、Beclin-1 mRNA及蛋白相对表达量均明显低于对照组,NOX2水平、NOX2 mRNA及蛋白相对表达量均明显高于对照组,差异均有统计学意义(P<0.05)。结论CaR通过TRPC6介导的自噬降低软骨细胞氧化应激水平,可能是延缓骨关节炎发生、发展的重要机制。