Neuroprotective effect of peroxiredoxin 6 against hypoxia-induced retinal ganglion cell damage

Background  

The ability to respond to changes in the extra-intracellular environment is prerequisite for cell survival. Cellular responses to the environment include elevating defense systems, such as the antioxidant defense system. Hypoxia-evoked reactive oxygen species (ROS)-driven oxidative stress is an underlying mechanism of retinal ganglion cell (RGC) death that leads to blinding disorders. The protein peroxiredoxin 6 (PRDX6) plays a pleiotropic role in negatively regulating death signaling in response to stressors, and thereby stabilizes cellular homeostasis.     

硒对钝顶螺旋藻氧化损伤的拮抗作用的光谱学特性及机制

研究了亚硒酸钠预处理对H2O2氧化胁迫下钝顶螺旋藻(Spirulina platensis)的生长、藻丝体形态、谱学特性以及细胞内活性氧(ROS)水平的影响,探究硒拮抗氧化胁迫保护螺旋藻的机制。结果显示,H2O2氧化胁迫明显抑制螺旋藻的生长,藻丝体严重受损,可见光吸收440nm峰增强,620和680nm峰降低;荧光发射和激发光谱特征峰强度明显降低,藻胆蛋白特征发射峰由660nm蓝移至650nm;红外光谱透射峰没有发生位移,蛋白质和多肽的特征谱带酰胺Ⅰ带和酰胺Ⅱ带相对强度降低;细胞内ROS相对含量显著性升高。硒预处理24h呈剂量效应地减轻由H2O2胁迫引起的氧化损伤,有效地抑制胞内ROS的过度累积,提高螺旋藻的抗氧化能力,缓解了氧化胁迫对光能捕获和传递等重要生理功能的影响。     

小鼠甲醇氧化应激的电子顺磁共振的研究

建立了小鼠甲醇中毒的氧化应激模型,研究了甲醇诱导小鼠产生氧化应激状态,探讨了大黄中草药和维生素C对小鼠甲醇氧化应激的保护作用. 模型小鼠共分为5个组：空白组、对照组、甲醇应激组、大黄组、维生素C（Vc）组. 利用电子自旋捕捉技术研究了小鼠甲醇氧化应激不同模型组小鼠体内的肝、肾、脾、心、肺和脑中产生的自由基强度的变化. 结果表明： 甲醇诱发了小鼠体内自由基的产生,甲醇组和生理盐水组相比,自由基的强度明显增强（显著性差异统计指标P<0.01);而大黄和Vc对小鼠甲醇氧化应激有保护作用,大黄组、Vc 组和甲醇组相比自由基的强度则明显降低（P<0.01).     

Variations of the antioxidant system during development of the cold-tolerant beetle, Tenebrio molitor

Data relating to the peculiarities of antioxidant enzyme activities, glutathione (GSH) level and lipid peroxidation (LP) intermediates specific to at each ontogenic stage of Tenebrio molitor are presented. Metamorphosis is accompanied by a shift of prooxidant-antioxidant balance towards oxidative processes, since pupae have the highest levels of lipid peroxidation intermediates. Cold acclimation (4 degree C) can promote oxidative stress at the cold sensitive developmental stage--imagoes, which enhance their levels of diene conjugates and ketodienes after a 2-week cold acclimation. This enhancement is accompanied by an increase in catalase (CAT) activity. GSH levels undergo no changes in imagoes after cold acclimation. Neither larval nor pupal T. molitor show significant changes in LP product contents after cold acclimation. Chilling results in a significant increase in CAT activities in pupae, but not in larvae. GSH levels are reduced both in larvae and pupae after cold exposure. However, cold acclimation does not affect superoxide dismutase (SOD) activity in any of the developmental stages of T. molitor.     

利用FCLA特征荧光实现单态氧和超氧的高灵敏检测

活性氧(ROS)的检测对生物学和医学研究很多领域方面具有重要意义,如光动力治疗肿瘤的治疗检测、植物应激反应检测、种子活力检测等。实现活性氧中的单态氧、超氧阴离子高灵敏度的检测,是人们追求的目标。研究发现单态氧、超氧的选择性化学发光探针FCLA(fluoresceinyl cypridina luciferin analog)氧化后,在515 nm处荧光会大大增强。利用这个性质并结合非常成熟的荧光检测方法,可以实现活性氧的高灵敏度检测。尤其在微观检测方面,如单细胞, FCLA的化学发光很微弱而难以检测到,但是却可以灵敏地检测到其荧光变化。FCLA荧光检测不仅灵敏度高而且选择性好,其无毒和易于透膜的性质更适于在细胞和生物体上应用。另外如果同时结合FCLA的化学发光,对单态氧和超氧阴离子的检测将更准确、更具实时性。     

Toxicity of nano- and micro-sized ZnO particles in human lung epithelial cells

This is the first comprehensive study to evaluate the cytotoxicity, biochemical mechanisms of toxicity, and oxidative DNA damage caused by exposing human bronchoalveolar carcinoma-derived cells (A549) to 70 and 420 nm ZnO particles. Particles of either size significantly reduced cell viability in a dose- and time-dependent manner within a rather narrow dosage range. Particle mass-based dosimetry and particle-specific surface area-based dosimetry yielded two distinct patterns of cytotoxicity in both 70 and 420 nm ZnO particles. Elevated levels of reactive oxygen species (ROS) resulted in intracellular oxidative stress, lipid peroxidation, cell membrane leakage, and oxidative DNA damage. The protective effect of N-acetylcysteine on ZnO-induced cytotoxicity further implicated oxidative stress in the cytotoxicity. Free Zn²⁺ and metal impurities were not major contributors of ROS induction as indicated by limited free Zn²⁺ cytotoxicity, extent of Zn²⁺ dissociation in the cell culture medium, and inductively-coupled plasma-mass spectrometry metal analysis. We conclude that (1) exposure to both sizes of ZnO particles leads to dose- and time-dependent cytotoxicity reflected in oxidative stress, lipid peroxidation, cell membrane damage, and oxidative DNA damage, (2) ZnO particles exhibit a much steeper dose–response pattern unseen in other metal oxides, and (3) neither free Zn²⁺ nor metal impurity in the ZnO particle samples is the cause of cytotoxicity.     

Cell Membrane-Coated AuNPs as a New Biomimetic Platform for ROS Scavenger Provides Cytoprotection from Apoptosis Induced by Oxidative Stress

Biocompatible nanoparticles have attracted considerable attention as cellular antioxidant agents for the improvement of new therapeutics for several and diverse oxidative stress related disorders. Here, it is shown that the biocompatible cell membrane@gold nanoparticle displays catalase-mimic behavior by decomposing H₂O₂. This interesting behavior can be used for cell protection against induced oxidative injury by reactive oxygen species (ROS). So, after characterizing this novel biocompatible nanostructure, the protective effect of cell membrane@AuNPs pretreatment in front of a well-defined oxidative stress-inducing chemotherapy drug, doxorubicin (Dox) is assessed. Doxorubicin pretreatment significantly reduces the intracellular ROS production. Similarly, a reduction in the levels of DNA oxidative damage, as measured with the AO/EB staining, is also observed. Obtained results would support that this novel nanostructure can show how a pharmacological agent can be used against oxidative stress-mediated diseases.     

Quantitative analysis of oxidative DNA damage induced by high-voltage pulsed discharge with cavitation

Pulsed discharge is used for sterilization and disinfection, but the details of the molecular mechanisms remain largely unknown. Since pulsed discharge generates reactive oxygen species (ROS), we analyzed the oxidative DNA damages after pulsed discharge treatment to consider the involvement of ROS in the damaging process. We applied pulsed discharge with cavitation to plasmid DNA molecules and estimated the yields of the damages by agarose gel electrophoresis. The treated DNA contained various oxidative DNA damages, including single and double strand breaks and base lesions. The yields of the damages increased in response to the energy used for pulsed discharge. We also measured the yield of 8-hydroxyguanine (8-OH-G), one of the major oxidative base lesions, in the treated plasmid DNA by mass spectrometry quantitatively and found that the yield of the oxidative base lesion corresponded to the increment of the applied energy. In addition, we observed the involvement of mutM gene, which is responsible for repair of 8-OH-G, in the increased sensitivity of Escherichia coli to pulsed discharge. Therefore, ROS seem to mediate the sterilization ability of pulsed discharge.     

Survival at low temperatures in insects: what is the ecological significance of the supercooling point?

Many freezing-intolerant insects may die during long or even brief exposures to temperatures above their supercooling point (SCP). Consequently, the real ecological value of the SCP remains ambiguous, particularly for tropical species that never experienced cold exposures. The bimodal distribution of SCP is discussed in the light of sexual dimorphism. The importance of sex in insect cold hardiness has been regularly neglected and although we admit that in some species sex may be uneasy to determine, it should be taken into account in further studies. We suggest that supercooling ability may be, at least partially, a result of adaptations to other functions unrelated to cold, including the desiccation resistance. The potential causes of insect death at low temperatures during survival experiments have also been examined. Prolonged exposures at lethal low temperatures can produce deleterious effects (including death) even if the insect does not freeze; during long-term exposure to low temperatures the organisms may finally die from the exhaustion of energy reserves.     

Gene expression in response to low temperatures in mammalian cells: a review of current ideas

This review was undertaken to identify responses of mammalian cells to cold temperatures, such as might be encountered in therapeutic procedures where body temperatures are lowered, in preservation of cells and organs, and in cryopreservation. In general, cold elicits a range of stress responses through identified signaling pathways, which may determine the survival or otherwise of the cells. Under conditions of mild hypothermia, there is evidence for responses which reflect an ordered acclimation to the new environment, whilst deep cooling invokes a more generalised stress response.     

New insights on the role of ROS in the mechanisms of sonoporation-mediated gene delivery

Reactive oxygen species (ROS) are hypothesized to play a role in the sonoporation mechanisms. Nevertheless, the acoustical phenomenon behind the ROS production as well as the exact mechanisms of ROS action involved in the increased cell membrane permeability are still not fully understood. Therefore, we investigated the key processes occurring at the molecular level in and around microbubbles subjected to ultrasound using computational chemistry methods. To confirm the molecular simulation predictions, we measured the ROS production by exposing SonoVue® microbubbles (MBs) to ultrasound using biological assays. To investigate the role of ROS in cell membrane permeabilization, cells were subjected to ultrasound in presence of MBs and plasmid encoding reporter gene, and the transfection level was assessed using flow cytometry. The molecular simulations showed that under sonoporation conditions, ROS can form inside the MBs. These radicals could easily diffuse through the MB shell toward the surrounding aqueous phase and participate in the permeabilization of nearby cell membranes. Experimental data confirmed that MBs favor spontaneous formation of a host of free radicals where HO was the main ROS species after US exposure. The presence of ROS scavengers/inhibitors during the sonoporation process decreased both the production of ROS and the subsequent transfection level without significant loss of cell viability. In conclusion, the exposure of MBs to ultrasound might be the origin of chemical effects, which play a role in the cell membrane permeabilization and in the in vitro gene delivery when generated in its proximity.     

Snow white and the seven dwarfs: a multivariate approach to classification of cold tolerance

Two main cold hardiness strategies of insects - freeze tolerance in some species, and overwintering in a supercooled state without tolerance of freezing in many others - were recently reclassified. However, I present several problems with the current systems. My suggested classification is based on clearer definitions of the causes of cold injury. I recognize three main mortality factors: freezing of body liquids, cold shock, and cumulative chill injury. Presence or absence of each of these factors produce eight combinations. I have named the eight classes after Snow White and the Seven Dwarfs to avoid nomenclatural confusion. Some of these classes are probably not used as tactics against cold injury by any insect species. Other classes contain so many species that they might be reclassified in more detail, using values of supercooling point and other quantitative parameters. However, widely comparable parameters, like the upper limit of cold injury zone and the sum of injurious temperatures are still rarely published, thus we still lack comprehensive data for multivariate analyses. Every cold hardiness strategy should be characterized by a meaningful class or subclass together with the physiological, biochemical, and behavioural mechanisms employed by the insects. I also point out the existence of strategies that combine two tactics - either a switching strategy (during preparation for winter, population "chooses" which tactic will be used), or a dual strategy (individuals are ready to use one of the tactics depending on the prevailing environmental conditions).     

在体电子自旋共振(EPR)波谱和成像技术在生物医学中的应用

活性氧和氮自由基(ROS/RNS)在一系列的人类疾病中扮演着双重角色. 它们可以是氧化剂, 诱导氧化状态, 导致组织损伤. 它们又可以是信号传导因子, 诱发保护性反应, 使得被调节的组织器官经受得起更强的损伤. 鉴于它们在生物医学中的重要作用, 检测它们产生和分布的技术的研究因而变得必要和紧迫. 在体电子自旋共振(EPR)波谱和成像技术渐已被应用于活体生物体系中用以表针和显像ROS/RNS. EPR 波谱特性(包括线宽、强度和寿命)以及空间分布信息已为动物甚至人体病理模型中氧化还原状态和氧分布的检测提供不可缺少的依据. 该文将简单描述和讨论一系列在体EPR 波谱和成像技术在器官和组织中的应用, 其中包括活体组织氧化还原状态, 活体组织氧分布和时间演化, 自由基空间以及谱-空间成像等.     

Research into accelerator driven systems using particle track detectors

In the interaction of relativistic protons with heavy and extended targets such as lead, large number of neutrons is produced in the course of the so-called spallation process. These neutrons can be used to drive a sub-critical nuclear assembly for energy generation and/or for the transmutation of the long-lived nuclear waste isotopes to environmentally safer nuclear species. Such nuclear assemblies are referred to as accelerator driven systems (ADS).

Knowledge of the neutron yield in the spallation process and an understanding of the behaviour of these neutrons in the desired sub-critical assembly are the most important and determining factors in the design and operation of these systems. Many parameters related to the neutronics of an ADS can be studied qualitatively as well as quantitatively using solid-state nuclear track detectors (SSNTD). In some circumstances SSNTDs provide the best and the most logical detector option for these investigations.

In this paper applications of the SSNTDs into research related to ADS are discussed and some experimental and theoretical results presented.     

The physics of accelerator driven sub-critical reactors

In recent years, there has been an increasing worldwide interest in accelerator driven systems (ADS) due to their perceived superior safety characteristics and their potential for burning actinides and long-lived fission products. Indian interest in ADS has an additional dimension, which is related to our planned large-scale thorium utilization for future nuclear energy generation. The physics of ADS is quite different from that of critical reactors. As such, physics studies on ADS reactors are necessary for gaining an understanding of these systems. Development of theoretical tools and experimental facilities for studying the physics of ADS reactors constitute important aspect of the ADS development program at BARC. This includes computer codes for burnup studies based on transport theory and Monte Carlo methods, codes for studying the kinetics of ADS and sub-critical facilities driven by 14 MeV neutron generators for ADS experiments and development of sub-criticality measurement methods. The paper discusses the physics issues specific to ADS reactors and presents the status of the reactor physics program and some of the ADS concepts under study.      

Expression and distribution of aquaporin 8 in rat hepatocytes cold stored 72 hours in modified University of Wisconsin and bes-gluconate-sucrose solutions. Study of their correlation with water content

Since few data are availble on the genetic responses to low temperatures, we investigated if cold storage of hepatocytes (0 degree C, mUW or BGS solutions, 72 h) can affect gene expression and/or cellular localization of AQP8 and their correlation with water movements. Cold preserved hepatocytes showed a significant decrease in water content (P less than 0.05) but were able to regulate their volume when they returned to physiological conditions. These changes were not related to modulation in the expression and the pattern of distribution of AQP8 suggesting that other mechanisms are involved. The study of the quantitative changes in the expression of genes coding for liver specific proteins in cold preserved hepatic cells is of interest in order to develop new preservation methods or solutions that could contribute to maintain the utility of these cells when destined to be applied in clinical models.     

Fluorogenic Probes Applied to the Study of Induced Oxidative Stress in the Human Leukemic HL60 Cell Line

Development of microspectrofluorometric methods using specific fluorogenic probes has provided precious help in studying in situ oxidative stress and cellular protective systems. The aim of this study was to determine ROS production concomitantly with a modification of the intracellular thiol pool after applying an oxidative stress to a nonadherent cell model represented by the HL60 cell line. The dichlorodihydrofluorescein diacetate (H₂DCFDA) probe assessed the kinetic production of ROS by cells submitted to the chemical oxidant t-butylhydroperoxide with a high signal/noise ratio. The probe sensitivity permitted us to detect endogenous ROS production in HL60 cells and the protective effect of N-acetyl cysteine against ROS. The chloromethylfluorescein diacetate probe (CMFDA) permitted us to evaluate the thiol depleting effect of N-ethyl maleimide. Complete thiol depletion was associated with a moderate increase in ROS production. The cell viability was determined with calcein-AM, which gave results similar to those with the tetrazolium dye. This probe was not affected by intracellular pH and did not required an extraction step, unlike tetrazolium dye. In conclusion, cell-permeant fluorogenic probes are useful and sensitive tools to determine in situ ROS production concomitantly with consecutive change in the thiol system in a living and non-adherent cell model.     

Effect of UV radiation on the optical and some structural properties of irregular rayon acetate fibers

This recent study shed demonstration on how the structural parameters changes due exposure to UV radiation and different wavelengths. So this article throws light on the changes of the optical properties and some structural properties due exposure to UV radiation and different wavelengths on irregular rayon acetate fibers. Pluta polarizing interference microscope is used to study the changes for refractive indices and birefringence with different exposure times at three different intensities on irregular rayon acetate fibers. As the principal optical parameters determined, these give an indication of the isotropic refractive index, stress optical coefficient, stress due to UV radiation, optical configuration parameter, segment anisotropy, the number of molecules per unit volume and polarizability of a monomer unit can be obtained. Also by using different wavelengths are given the spectral dispersions, Cauchy's dispersion constants, the resonant wavelength, the oscillation energy, the dispersion energy, the dielectric constant and the dielectric susceptibility for rayon acetate fibers with different wavelengths at room temperatures. Illustrations are given using graphs and microinterferograms.