铜锌超氧物歧化酶与过氧化氢反应中羟自由基的形成

应用脱氧核糖降解法研究了离体条件下Cu,Zn-SOD与H2O2反应产生·OH,并对其机理进行了探讨。H2O2可使Cu,Zn-SOD失活,在失活过程中有·OH产生,甲酸钠和苯甲酸钠均能不同程度地保护Cu,Zn-SOD和降低H2O2与Cu,Zn-SOD反应中·OH的产额;热失活SOD也可和H2O2反应生成·OH,且效能高于活性Cu,Zn-SOD;用螫合剂脱去Cu,Zn-SOD的金属辅基后,脱辅基的SOD蛋白不能和H2O2反应产生·OH;Cu2＋和H2O2反应产生·OH的效率很高,而Zn2＋产生·OH的效率很低。实验结果提示Cu,Zn－SOD与H2O2反应产生的·OH可能是SOD活性中心的Cu2＋与H2O2发生Fenton反应的结果．     

辐射及活性氧对DNA的损伤以及芥子碱的保护作用

在Ｘ射线照射下,小牛胸腺ＤＮＡ的碱基损伤及链断裂随着剂量升高而增加,其损伤主要集中于链断裂;活性氧可以引起ＤＮＡ损伤,Ｈ２Ｏ２仅造成少量伤害,当在含有Ｈ２Ｏ２的体系中加入微量的Ｃｕ＾２＋、Ｆｅ＾２＋时损伤急剧增加,这是由反应产生的．ＯＨ所致,Ｃｕ＾２＋的致损伤效果明显高于Ｆｅ＾２＋。ＯＨ清除剂芥子碱具有很强的抗辐射及抗氧化作用,且对ＤＮＡ无伤害。这说明ＯＨ在ＤＮＡ的氧化损伤中起重要作用。     

渥曼青霉素对端粒DNA链断裂重连接作用的影响

端粒是位于真核细胞染色体末端的DNA-蛋白质复合体,在维持染色体稳定上起着重要的作用,并且与细胞的衰老、癌变有着密切的关系。本实验观察了DNA依赖性蛋白激酶(DNA-dependent protein kinase,DNA-PK)抑制剂-渥曼青霉素(wortmannin,WM)对H2O2诱导的HeLa细胞端粒DNA链断裂重连接效应。结果表明WM能够显著地抑制H2O2诱导的HeLa细胞端粒DNA链断裂后的重连接作用,提示DNA-PK参与了端粒DNA链断裂损伤的修复过程。     

3-氨基苯酰胺对氧化应激诱导的HeLa细胞端粒DNA链断裂重连接作用的影响

端粒是位于真核细胞染色体末端的DNA-蛋白质复合体,在维持染色体稳定上起着重要的作用,并且与细胞的衰老和凋亡有着密切的关系.在各种DNA损伤中,单链断裂(single-strand breaks, SSBs)是最常见的类型之一,既可直接通过内源活性氧或离子化辐射产生,也可间接地在DNA代谢或碱基切除修复期间产生.已知多聚(ADP-核糖)聚合酶［poly(ADPribose) polymerase, PARP］在SSBs修复中起着极为重要的作用.本实验观察了PARP抑制剂3-氨基苯酰胺(3-aminobenzamide, 3-AB)对氧化应激诱导的HeLa细胞端粒DNA链断裂重连接的效应以及对过氧化氢(H2O2)抑制HeLa细胞增殖的影响.结果表明3-AB能够显著地抑制氧化应激诱导的HeLa细胞端粒DNA链断裂后的重连接作用,并能增强H2O2对HeLa细胞增殖的抑制作用,提示PARP参与了端粒DNA链断裂损伤的修复过程.     

泥鳅多糖清除活性氧和保护DNA链的作用

采用化学发光法和分光光度法在多种化学模拟体系中研究了泥鳅多糖清除活性氧的作用 ,并用化学发光法观察了泥鳅多糖对·OH导致DNA链损伤的抑制作用。结果表明 ,泥鳅多糖能够有效地清除O·-2 、·OH、H2 O2 等活性氧 ,对DNA链具有良好的保护作用     

夏枯草色素的抗氧化活性研究

目的:对夏枯草色素的抗氧化作用进行研究.方法:以蒸馏水为溶剂,从夏枯草中提取色素;采用比色法测定夏枯草色素对DPPH·、O2-·和·OH 3种自由基的清除作用以及还原能力.结果:夏枯草色素对三种自由基都有明显的清除作用,清除能力为DPPH·＞·OH＞ O2-·;对Fe3+也具有较强的还原能力.结论:清除率与浓度存在明显的量效关系,当色素浓度为0.13 mg/mL时,其清除率O2-·为4.1％、DPPH·为28.3％、·OH为17.9％;还原Fe3+能力随色素的浓度增加而增强.     

氧化胁迫下稻苗体内积累的脯氨酸的抗氧化作用

水稻幼苗在外源·OH和1O2氧化胁迫下,体内脯氨酸明显积累,而和H2O2处理对稻苗体内脯氨酸含量无影响。强光亦能诱导稻苗体内脯氨酸积累。脯氨酸预处理则显著抑制·OH、1O2及强光所诱导的稻苗膜脂过氧化作用。在活性氧的产生／检测系统中,脯氨酸对·OH和1O2所引发反应有明显的竞争效应,但对和H2O2的作用没有形响。这些都表明,氧化胁迫下稻苗体内积累的脯氨酸具有抗氧化作用;脯氨酸对活性氧的清除有一定的专一性,即只对·OH和1O2有明显的清除活性。     

沙棘果皮多糖清除氧自由基的活性研究

沙棘(Hippophae rhamnosides)果皮经80℃恒温水浴提取,乙醇沉淀得粗多糖.Sevag法去蛋白,经50%和70%乙醇分级,得3种级分沙棘多糖H1、H2和H3;以Fenton反应,即H2O2/Fe2 /水杨酸为·OH产生和检测体系;以邻苯三酚/EDTA/Tris-HCl为O2-.产生体系,对沙棘多糖H1、H2和H3进行抗氧自由基活性研究.结果表明,沙棘多糖对·OH和O2-.有较显著的清除能力.不同级分多糖H1、H2和H3浓度达200μg·mL-1时,对·OH的清除率分别为44.9%、49.0%和26.4%,抗O2-.活性分别为36.9%,15.4%和23.1%.多糖质量浓度增大时,两种自由基清除率增加,且呈量效关系.     

杨桃提取物体外清除氧自由基作用

从杨桃果中提取得到三种提取物为匀浆提取物、蛋白提取物和多糖提取物。采用化学发光法测定这三种提取物清除氧自由基的活性,实验结果:匀浆提取物清除羟自由基(·OH)和H2O2的活性大小相近,而清除超氧阴离子自由基(O2–·)的活性较小,其IC50约为前两者的4倍。蛋白提取物清除O2–·和·OH的活性大小相近,而清除H2O2的活性明显小于前两者,IC50约为前两者的9倍。多糖提取物清除.OH的活性明显大于清除O2–·和H2O2的活性,其IC50约为O2–·的1/22,约为H2O2的1/65。结果表明,杨桃果具有清除O2–·、·OH和H2O2的作用,不同提取物对这些活性氧自由基的清除能力有所不同。     

强光及外源活性氧对莴苣叶绿素荧光的影响

莴苣叶绿体在强光处理下发生先抑制,主要表现为叶绿素荧光参数Fv／Fm和ΦPSⅡ降低;外源活性氧H2O2、O2·OH和1O2均能引起叶绿体PSⅡ光化学效率Fv／Fm不同程度的下降,其中以1O2影响最明显;H2O2在诱导叶绿体荧光猝灭过程中,引起荧光产量降低,而使qp、qN、ΦPSⅡ、KD上升;在H2O2诱导的叶绿体荧光猝灭过程中,Fe2 能使qN和KD低于对照;由于1O2的产生对PSⅡ反应中心造成了损伤,引起qp和ΦPSⅡ下降。     

Oxidation of NADH by chloramines and chloramides and its activation by iodide and by tertiary amines

Irreversible oxidation of reduced nicotinamide nucleotides by neutrophil-derived halogen oxidants (HOCl, chloramines, HOBr, etc.) is likely to be a highly lethal process, because of the essential role of NAD(P)H in important cell functions such as mitochondrial electron transport, and control of the cellular thiol redox state by NADPH-dependent glutathione reductase. Chloramines (chloramine-T, NH(2)Cl, etc.) and N-chloramides (N-chlorinated cyclopeptides) react with NADH to generate the same products as HOCl, i.e., pyridine chlorohydrins, as judged from characteristic changes in the NADH absorption spectrum. Compared with the fast oxidation of NADH by HOCl, k approximately 3 x 10(5) M(-1) s(-1) at pH 7.2, the oxidation by chloramines is about five orders of magnitude slower; that by chloramides is about four orders of magnitude slower. Apparent rate constants for oxidation of NADH by chloramines increase with increasing proton or buffer concentration, consistent with general acid catalysis, but oxidation by chloramides proceeds with pH-independent kinetics. In presence of iodide the oxidation of NADH by chloramines or chloramides is faster by at least two orders of magnitude; this is due to reaction of iodide with the N-halogen to give HOI/I(2), the most reactive and selective oxidant for NADH among HOX species. Quinuclidine derivatives (QN) like 3-chloroquinuclidine and quinine are capable of catalyzing the irreversible degradation of NADH by HOCl and by chloramines; QN(+)Cl, the chain carrier of the catalytic cycle, is even more reactive toward NADH than HOCl/ClO(-) at physiological pH. Oxidation of NADH by NH(2)Br proceeds by fast, but complex, biphasic kinetics. A compilation of rate constants for interactions of reactive halogen species with various substrates is presented and the concept of selective reactivity of N-halogens is discussed.     

Polarity induced by chloramphenicol and relief by suA

The suA allele, known to relieve polarity in Escherichia coli, also relieves a unique polar effect on E. coli tryptophan operon messenger RNA produced by chloramphenicol.     

Hypobromous acid and bromamine production by neutrophils and modulation by superoxide

MPO (myeloperoxidase) catalyses the oxidation of chloride, bromide and thiocyanate to their respective hypohalous acids. We have investigated the generation of HOBr by human neutrophils in the presence of physiological concentrations of chloride and bromide. HOBr was trapped with taurine and detected by monitoring the bromination of 4-HPAA (4-hydroxyphenylacetic acid). With 100 microM bromide and 140 mM chloride, neutrophils generated HOBr and it accounted for approx. 13% of the hypohalous acids they produced. Addition of SOD (superoxide dismutase) doubled the amount of HOBr detected. Therefore we investigated the reaction of superoxide radicals with a range of bromamines and bromamides and found that superoxide radicals stimulated the decomposition of these species, with this occurring in a time- and dose-dependent manner. The protection afforded by SOD against such decay demonstrates that these processes are superoxide-radical-dependent. These data are consistent with neutrophils generating HOBr at sites of infection and inflammation. Both HOBr and bromamines/bromamides have the potential to react with superoxide radicals to form additional radicals that may contribute to inflammatory tissue damage.     

Pea seedling growth and development regulated by cotyledons and modified by irradiance

Growth and development of hydroponically grown pea seedlings ( Pisum sativum L. cv. Alaska) were measured using stem and root length as well as number of leaves and lateral roots. The growth was dependent on the presence of cotyledons and was modulated by the irradiance. All plants were grown in a full nutrient solution. If grown at low irradiance (73 μmol m^-2s^-1) they depended more and for a longer time on the cotyledons than plants grown at high irradiance (220 μmol m^-2s^-1). Low irradiance caused stem elongation but decreased root length and number of lateral roots as compared to plants grown at high irradiance. The dark respiration of the leaves was measured as oxygen uptake. In plants grown at the low irradiance, excision of the cotyledons caused the rate of oxygen uptake to increase by a factor of three, and the increase was sensitive to cyanide. Decotyledonized plants showed a high respiration rate and a diminished leaf growth for their entire life cycle. CO₂ fixation also increased in decotyledonized pea seedlings grown at either irradiance. The mobilization of food reserves from the seeds was positively correlated to seed dry weight, but only if the plants were grown at 73 μmol m^-2s^-1. Increasing dry weight of the seed enhanced top growth, whereas root growth was depressed, so that top and root responds differently with regard to that part of growth which depends on mobilization of reserves from the seed.     

Micronutrient Cation Sorption by Roots and Uptake by Plants

The sorption of ferric iron, copper, zinc and manganese by wheatseedling roots and by discs of cellulose filter paper was measured.The magnitude of sorption at pH 5-0 was Fe(III) > Cu(II)> Zn(II) > Mn(II). Sorption of Cu(II), Zn(II) and Mn(II)increased with increasing pH whilst sorption of Fe(III) decreased.The patterns of sorption are discussed in the light of our knowledgeof the hydrolysis of the metal ions. It is suggested that metalsadsorbed on root surfaces may be remobilized by organic ligandswhich leak from the root cells. Where an external liquid diffusionpath away from the root does not exist, soluble metal ligandcomplexes might accumulate in the water free space and superficialwater film of the root, thus facilitating their uptake intoroot cells and translocation within the plant. Under such conditionsthe amounts of metal translocated to the shoots of wheat seedlingsare shown to be related to the amounts of metal adsorbed bytheir roots. Key words: Adsorption, Micronutrients, Roots     

Diaphragmatic displacement measured by fluoroscopy and derived by Respitrace

In eight healthy volunteers we simultaneously measured the axial diaphragmatic motion by fluoroscopy and the cross-sectional area changes of the rib cage (RC) and abdomen (ABD) by Respitrace (RIP) during semistatic vital capacities (VC). We found that, if the fluoroscopic axial displacement of the posterior part of the diaphragm between residual volume (RV) and total lung capacity (TLC) is considered equal to 100%, the movement of the middle part is 90%, whereas that of the anterior part is only approximately 60%; the ratio of the axial displacements to mouth volume, furthermore, decreases at high lung volumes, especially for the anterior part. The RIP signal is nearly linearly related to mouth volume, but the contribution of the RC (delta RC) progressively increases (and is approximately 80% RIP at TLC), whereas the volume contribution of the ABD (delta ABD) levels off (to 20% RIP at TLC). The diaphragmatic volume displacement calculated from the theoretical analysis described by Mead and Loring also levels off at high volumes similarly as the ABD but is approximately 50% RIP at TLC. Finally, the axial movements of the three parts of the diaphragm are linearly related to the RC and ABD cross-sectional-area changes (r 0.91-0.97) and are even significantly better correlated with the "calculated" diaphragmatic volume displacement.     

Plaque Formation by Mumps Virus and Inhibition by Antiserum

Boston and ABC strains of mumps virus produced plaques approximately 1.0 mm in diameter in monolayers of BGM cells. The plaques were circular and either clear or target-like in form. Ricki strain virus produced plaques of similar size and form but, in addition, a red plaque was observed with this agent. The vaccine strain of mumps virus, Jeryl Lynn, produced minute clear plaques approximately 0.3 mm in diameter. Incorporation of diethylaminoethyl (DEAE)-dextran into the overlay medium did not affect the size difference between Jeryl Lynn plaques and those of the other strains. However plaques of the Jeryl Lynn and Ricki strains were more easily visualized when the overlay medium contained 400 mug/ml of DEAE-dextran. Simultaneous titration by plaque formation and roller tube infectivity showed that these two methods were of equal sensitivity. Virus neutralization by antibody was demonstrated by plaque reduction. Rise in antibody titer was observed in sera from human and animal infection, human vaccination, and rabbit immunization.