金电极上Cu,Zn-SOD的表面增强拉曼光谱

应用原位共焦拉曼光谱技术在经表面增强处理后的金电极上,观察Cu,Zn-SOD分子的吸附方式及其随电位变化的过程。实验结果表明,在400mV-700mV的电位区间内,Cu,Zn-SOD分子始终以氨基酸残基上COO-和NH2在电极表面吸附,使活性中心所处的平面远离电极表面,且随电位负移发生脱附。上述结果为解释SOD分子在金电极表面无法实现快速电子传递提供了直接依据。     

巯基乙胺修饰电极法测定植物体内超氧阴离子自由基

盐酸羟胺与超氧阴离子自由基反应生成NO2-,通过检测NO2-的生成量,可间接测定超氧阴离子自由基。基于上述原理,建立了一种用巯基乙胺自组装修饰金电极测定生物体系中超氧阴离子自由基的新方法。实验结果表明,在pH4.8的HAc-NaAc缓冲溶液中,修饰电极对NO2-的氧化具有良好的电催化作用,差分脉冲溶出伏安法测定其氧化峰电流与NO2-的浓度在5.0×10-8~1.0×10-4mol/L范围内呈线性关系,其线性回归方程为ipa(μA)=3.726C(μmol/L) 0.0257,相关系数为0.9983;检出限为1.0×10-8mol/L。该法用于玉米幼苗和叶片中超氧阴离子自由基产生速率的测定,结果令人满意。     

稳定直线型硝酮-O_2~(-·)加合物的构型因素解析

对生命体系中的氧自由基,Spin trapping-ESR方法是公认的特征分析手段[1].然而,对超氧阴离子自由基(O2-·)而言,其自由基加合物稳定性差.对此,人们通过改变硝酮的取代基团,设计出更为有效的O2-·捕获探针[2,3].另外,Zweier研究组[4,5]还应用密度泛函理论研究了影响五元环状硝酮     

New briaranes from the octocorals Briareum excavatum (Briareidae) and Junceella fragilis (Ellisellidae)

Six 12-hydroxybriaranes, including four new diterpenoids, briaexcavatins I-L (1-4), and two known metabolites, excavatolides C (5) and E (6), have been isolated from the cultured scleraxonia Briareum excavatum. In addition, the gorgonian coral Junceella fragilis yielded a new chlorinated briarane, fragilide C (10). The structures of above compounds were determined by spectroscopic methods and the structures of 5 and 6 were further confirmed by X-ray data analysis for the first time. The absolute configuration of 6 was elucidated by chemical conversion. Some of these briaranes have displayed inhibitory effects on superoxide anion generation by human neutrophils.     

Development of a new biosensor for superoxide radicals

A superoxide dismutase (SOD) biosensor for determination of superoxide radicals has been developed by immobilization of superoxide dismutase within gelatin (G) on a Pt electrode surface. The properties of the biosensor have been investigated and optimum conditions–enzyme concentration, glutaraldehyde concentration, and pH–were determined. The response of the G-SOD biosensor was proportional to concentration and the detection limit was 0.01 mmol L⁻¹ at a signal-to-noise ratio of 3. The biosensor retained 89% and 60% of its sensitivity after use for three and four weeks, respectively. Immobilization of SOD on gelatin provides a biocompatible microenvironment around the enzyme and stabilizes the activity of the enzyme very efficiently. The superoxide dismutase biosensor was used to determine the antioxidant properties of acetylsalicylic acid-based drugs and the anti-radical activity of healthy and cancerous human brain tissues.     

Effect of hydrostatic pressure on the fluorescence of indole derivatives

Effects of hydrostatic pressure on the fluorescence emission of L-tryptophan, N-acetyl-L-trytophanamide and indole were investigated. An increase in pressure ranging from 1 bar to 2.4 kbar results in reversible red-shifts of the emission of the three fluorophores. The pressure-induced redshift amounts to about 170 cm^–1 at 2.4 kbar, and appears related to changes in Stokes shift of the fluorophores caused by pressure effects on the dielectric constant and/or refractive index of the medium. As the pressure range investigated here is the range commonly used in studies of protein subunit association and/or folding, these observations raise the need for caution in interpreting pressure-induced spectral shifts. The significance of these observations to pressure studies of proteins is illustrated by investigation of pressure effects on human Cu,Zn Superoxide dismutase (SOD) and azurin fromPseudomonas aeruginosa. A reversible 170 cm^–1 red-shift of the emission of SOD was observed upon pressurization to 2.4 kbar. This might be interpreted as pressure-induced conformational changes of the protein. However, further studies using SOD that had been fully unfolded by guanidine hydrochloride, and fluorescence anisotropy measurements indicated that the observed red-shift was likely due to a direct effect of pressure on the fluorescence of the single tryptophan residue of SOD. Similar pressure-induced red-shifts were also observed for the buried tryptophan residue of azurin or for azurin that had been previously denatured by guanidine hydrochloride. These observations further suggest that the effective dielectric constant of the protein matrix is affected by pressure similarly to water.     

Sonochemistry of cytochrome c. Evidence for superoxide formation by ultrasound in argon-saturated aqueous solution

Formation of superoxide anion radicals (O₂^·−) induced by 50 kHz ultrasound in argon (Ar)-saturated aqueous solution was studied. Although EPR-spin trapping study with 5,5-dimethyl-1-pyrroline N-oxide (DMPO;100 mM) revealed the formation of DMPO-adducts of hydroxyl radicals (^·OH) and hydroperoxy radicals (HO₂^·; acid form of O2^·−) in O₂-saturated solution after sonication, no evidence of HO₂^· was found in Ar-saturated solution. When ferricytochrome c (cytochrome c) in Ar-saturated aqueous solution was sonicated, the reduced form of cytochrome c was observed and 80% of its formation was inhibited by the addition of superoxide dismutase (SOD). Sodium formate enhanced the production of the reduced form of cytochrome c. The % inhibition by SOD for the reduction increased in the order of Xe > Ar > He in accord with the higher temperatures of the cavitation bubbles. These results indicate that the O₂^·− is formed by directly by the sonolysis of water in the absence of O₂ when the temperature of cavitation bubble collapse is sufficiently high.     

超氧阴离子

介绍了超氧阴离子 O-·2 的产生、检测、化学性质、反应动力学、生物学研究、应用等方面的研究进展。     

用清除超氧阴离子自由基法评价竹叶提取物抗氧化能力

通过对邻苯三酚反应体系的吸收光谱,对自氧化速率及邻苯三酚浓度和缓冲液pH值对反应体系产生的超氧阴离子自由基清除率的影响进行了研究.分光光度法测定邻苯三酚反应体系的检测波长为319.5眦,反应体系体积10 mL,反应时间为9 min,3 mmol·L-1邻苯三酚加入量为0.3 mL,Tris-HCl缓冲液pH值为8.2,自氧化速率为0.035;用上述方法研究合成抗氧化剂叔丁基对苯二酚(TBHQ)和竹叶提取物样品浓度和清除率的关系,以IC50值(清除率为50%时的浓度值)作为评价指标,测得TBHQ和效果最好样品的IC50值分别为TBHQ(95.01 mg·L-1)和M40(298.69 mg·L-1),M40等竹叶提取物可以作为天然抗氧化剂进行开发.