Changes in antioxidative system and membrane damage of lettuce in response to salinity and boron toxicity

Individual and combined effects of salinity and B toxicity on growth, the major antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; ascorbate peroxidase, APX) activities, ascorbic acid, proline, and H₂O₂ accumulation, and stomatal resistance (SR), malondialdehyde (MDA), membrane permeability (MP) and the concentrations of sodium (Na), chloride (Cl) and boron (B) of lettuce were investigated. Boron toxicity and salinity reduced growth of lettuce plants. Under B toxicity, B concentration of the plants was increased, but in the presence of NaCl, the concentration of B was significantly reduced. Sodium and Cl concentrations were increased in B + NaCl and NaCl treatments. Membrane damage was more pronounced in NaCl and B + NaCl treatments. Stomatal resistance of the plants was significantly increased by salinity treatments. The accumulation of proline and ascorbic acid was the highest in the B + NaCl treatment. In general, stress conditions significantly increased H₂O₂ and antioxidant enzyme (SOD, CAT and APX) activities. The present results indicate that stomatal closure is an important response of lettuce against NaCl and B + NaCl stress. Furthermore NaCl and B + NaCl toxicity-induced oxidative stress in lettuce resulting in lipid peroxidation and membrane damage. Increased antioxidant enzyme activities and also accumulation of ascorbic acid and proline are involved in order to overcome B- and NaCl-induced oxidative stress.     

First Report of a Peroxiredoxin Homologue in Jellyfish: Molecular Cloning,Expression and Functional Characterization of CcPrx4 from Cyanea capillata

We first identified and characterized a novel peroxiredoxin (Prx), designated as CcPrx4, from the cDNA library of the tentacle of the jellyfish Cyanea capillata. The full-length cDNA sequence of CcPrx4 consisted of 884 nucleotides with an open reading frame encoding a mature protein of 247 amino acids. It showed a significant homology to peroxiredoxin 4 (Prx4) with the highly conserved F-motif (⁹³FTFVCPTEI¹⁰¹), hydrophobic region (²¹⁷VCPAGW²²²), ¹⁴⁰GGLG¹⁴³ and ²³⁹YF²⁴⁰, indicating that it should be a new member of the Prx4 family. The deduced CcPrx4 protein had a calculated molecular mass of 27.2 kDa and an estimated isoelectric point of 6.3. Quantitative real-time PCR analysis showed that CcPrx4 mRNA could be detected in all the jellyfish tissues analyzed. CcPrx4 protein was cloned into the expression vector, pET-24a, and expressed in Escherichia coli Rosetta (DE3) pLysS. Recombinant CcPrx4 protein was purified by HisTrap High Performance chelating column chromatography and analyzed for its biological function. The results showed that the purified recombinant CcPrx4 protein manifested the ability to reduce hydrogen peroxide and protect supercoiled DNA from oxidative damage, suggesting that CcPrx4 protein may play an important role in protecting jellyfish from oxidative damage.     

ROS对氯化镉诱导的人乳腺癌MCF-7细胞凋亡的影响

为研究活性氧(ROS)对氯化镉(CdCl2)诱导的人乳腺癌MCF-7细胞凋亡的影响,本文采用荧光探针DHR检测CdCl2诱导的MCF-7细胞内ROS的变化水平;分别采用MTT法和DNA Ladder法检测CdCl2对细胞的毒性作用和细胞DNA的凋亡情况;通过加入自由基清除剂LNAC抑制细胞内的ROS水平,从而研究ROS对细胞凋亡的影响。结果显示,1.00×10-3 mol/L的CdCl2处理MCF-7细胞6h,细胞内ROS水平为对照组的2.04倍,细胞存活率为26.54%,DNA出现片段化分解。用CdCl2+LNAC复合处理MCF-7细胞6h,细胞内ROS水平为对照组的1.25倍,细胞存活率为61.17%,DNA不出现凋亡条带。因此,由CdCl2所导致的MCF-7细胞凋亡和DNA片段化分解,与细胞内ROS水平的升高有关。     

漆酶活化处理条件对竹材活性氧类自由基变化的影响

采用电子自旋共振波谱检测技术,分析漆酶活化毛竹材产生的自由基种类,并结合单因素方差分析方法,分析漆酶活化处理条件对毛竹材产生的活性氧类自由基相对浓度的影响。未处理毛竹材产生的自由基属酚氧自由基,而漆酶处理毛竹材产生的自由基属典型的活性氧类自由基。酶用量、反应体系pH值、处理时间、处理温度及Cu~(2 )浓度,均对活性氧类自由基相对浓度有极显著影响。当采用酶用量为30u/g、反应体系pH值为4、处理时间为2h、处理温度为60℃、Cu~(2 )浓度为30mmol/L时,漆酶活化毛竹材产生的活性氧类自由基的相对浓度最高。     

线粒体中活性氧的产生途径和测定方法

活性氧(ROS)是自然界普遍存在的含氧化合物的总称,对生物体的生理和细胞稳态的调节具有重要意义.但活性氧的测定方法仍然是生物研究领域的一个难点,本文综述了线粒体中活性氧的产生途径和测定方法.     

Differential response of zucchini varieties to low storage temperature

Several parameters related to cold damage in zucchini have been measured in fruits from different varieties cultivated in the province of Almería (south-eastern Spain). The purpose of this work is to correlate physiological parameters with commercial quality and chilling damage, in order to establish reliable criteria for selecting varieties having better genetic adaptation to low-temperature storage. The results show that, for varieties harvested during February, the variety Natura was the best adapted to storage at 4 °C, as its fruits suffered less from chilling injuries, had a lower weight loss, and the levels of metabolites malonyldialdehyde and H₂O₂ were not as high as in the other varieties. Genetic variation was detected for some of the parameters analysed. The positive correlation among the levels of MDA, H₂O_2, and chilling injury, as well as the negative correlation among catalase and chilling injury, makes these parameters good indicators of chilling damage in our system.     

Stimulation of ROS-scavenging systems in squash (Cucurbita pepo L.) plants by compost supplementation under normal and low temperature conditions

The beneficial effect of compost, the final product of aerobic biodegradation of organic matter, on growth, lipid peroxidation [as malondialdehyde (MDA], hydrogen peroxide (H₂O₂) and superoxide anion (O₂^•−), activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR), as well as reduced ascorbate (ASC) and glutathione (GSH) and their oxidized forms was investigated in squash (Cucurbita pepo L. cv. Eskandarany) plants exposed to normal and low temperature (LT) conditions. LT stress of 8 °C significantly reduced the plant growth of untreated plants, but compost alleviated the adverse effect of stress and significantly increased the fresh and dry weights under normal and stress conditions. LT also induced accumulation of H₂O₂ and O₂^•− and resulted in increased lipid peroxidation, pointing out to cellular oxidative stress. Under compost application, such reactive oxygen species (ROS) and peroxidized lipids were markedly reduced, but SOD, CAT, APX and GR activities, key enzymes of ROS-scavenging systems, were significantly increased. Data also indicated that there were general reductions in total ascorbate and glutathione pool in LT control plants, but compost-treated ones considerably have maintained higher levels of such redox metabolites. Significantly higher ratios of ASC/DHA (dehydroascorbate) and GSH/GSSG (glutathione disulfide) were generally found in compost-treated plants than in untreated-ones. It is evident that compost induced enhancement of LT tolerance was related to up-regulation of enzymatic and non-enzymatic antioxidant systems. Such enhancement would eventually protect plant cells from LT-induced oxidative stress reactions via scavenging ROS.     

Hepatoprotective activity of angiotensin-converting enzyme (ACE) inhibitors, captopril and enalapril, against paraquat toxicity

Paraquat is a highly toxic herbicide that is used in most countries without restriction. The cytotoxic action of paraquat is mediated by reactive radicals that are products of its metabolic reduction in cells. It has already been hypothesized that some angiotensin-converting enzyme inhibitors (e.g., captopril and enalapril) could show antioxidant and radical scavenging activity through their structural thiol groups, increasing antioxidant enzymes production or nitric oxide synthesis. In this study the hepatoprotective effect of captopril and enalapril against paraquat induced oxidative stress cytotoxicity was evaluated in isolated rat hepatocyte. Subtoxic concentrations of captopril (0.2 mM) and enalapril (0.2 mM) significantly (p < 0.05) protected the hepatocytes against paraquat (2 mM) induced oxidative stress cytotoxicity markers including: cell lysis, reactive oxygen species (ROS) generation, lipid peroxidation, glutathione depletion, mitochondrial membrane potential decrease, lysosomal membrane oxidative damage and cellular proteolysis. Moreover, we showed that non-thiol enalapril acts as well as thiol containing captopril at inhibiting oxidative stress cytotoxicity markers. Finally, our results support the hypothesis that it is the increase in nitric oxide synthesis and not the presence of the thiol group that accounts for the antioxidant activity of ACE inhibitors.     

In vitro effect of recombinant human tumor necrosis factor-alpha on canine neutrophil apoptosis

Neutrophils (polymorphonuclear leukocytes: PMNs) are essential for the host defense against various infections and are often injurious to the host, causing inflammatory diseases where tumor necrosis factor-alpha (TNF-alpha) is suggested to play an important role. Since an effect of TNF-alpha on canine PMN apoptosis has not been studied, canine PMNs were stimulated with recombinant human (rh)TNF-alpha in the present study to investigate the effect of TNF-alpha on canine PMN apoptosis. PMN apoptosis and function to produce ROS were assessed by flow cytometry. Delayed apoptosis was observed in the PMNs treated with rhTNF-alpha at 100 ng/ml, accompanied by retention of capability to produce ROS. However, PMN apoptosis was accelerated by rhTNF-alpha combined with cycloheximide. Therefore, it is indicated that TNF-alpha is able to activate anti- and pro-apoptotic pathways in PMNs and that the inhibition of PMN apoptosis by TNF-alpha requires protein synthesis in the PMNs.     

Effect of antioxidant supplementation on free radical scavenging system and immune response in lindane treated scabies patients

The scabicide, lindane induces oxidative stress and immunological alterations. The present study was undertaken to assess the ameliorative effects of antioxidant supplementation in lindane treated scabies patients. Scabies patients were treated with either 1% lindane or 1% lindane along with antioxidant (Lycored or Vitamin-E). Oxidative stress and immunological parameters were evaluated in blood samples and compared with healthy controls. Lindane caused a significant increase in malonedialdehyde (MDA) levels and decrease in reduced glutathione (GSH) and ferric reducing ability of plasma (FRAP), which was attenuated by anti-oxidant therapy. The IL-1α levels were significantly enhanced in scabies patients per se and remained unaffected after lindane/anti-oxidant treatment. The TNF-α and nitroblue tetrazolium (NBT) reduction levels were not significantly different in all the groups. Topical application of lindane induces significant free radical generation and may cause immunological alterations which can be reversed by antioxidant therapy.