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Cell Localization of DPI-Dependent Production of Superoxide in Reproductive Tissues of the Olive Tree (Olea europaea L.) 橄榄树(Olea europaea L.)生殖组织中DPI依赖性超氧化物产生的细胞定位
Pub Date : 2022-04-19 DOI: 10.3390/oxygen2020007
M. J. Jiménez-Quesada, A. Castro, E. Lima-Cabello, J. Alché
Reactive Oxygen Species (ROS) are compounds derived from oxygen with important implications in biological processes in plants, some of them related to reproduction. Among ROS, superoxide is the primary oxidant, since an array of other ROS are eventually derived from this anion. Therefore, analysis of the molecular systems able to generate this molecule and the cellular compartmentalization of these events is of paramount importance. We have used the fluorochrome DCFH2-DA and the chromogenic substrate NBT in association with DPI (a specific inhibitor of Rboh enzymes generating superoxide in plants) in combination with confocal microscopy and stereomicroscopy, respectively to identify cell localization of ROS in general, and superoxide accumulation in olive reproductive tissues. A significant production of both ROS and superoxide has been described, showing a fairly precise spatial and temporal location throughout olive flower development. The reduction of the NBT signal after the addition of DPI suggests that the generation of superoxide is largely due to Rboh or other flavin oxidase activity. At the subcellular level, accumulation of O2●− has been located in the plasma membrane of mature pollen and germinated pollen, as well as in the rough endoplasmic reticulum and in mitochondria.
活性氧(Reactive Oxygen Species, ROS)是由氧衍生的化合物,在植物的生物过程中具有重要意义,其中一些与生殖有关。在活性氧中,超氧化物是主要的氧化剂,因为一系列的其他活性氧最终来源于这个阴离子。因此,分析能够产生这种分子的分子系统和这些事件的细胞区隔是至关重要的。我们利用荧光染料DCFH2-DA和与DPI(植物中产生超氧化物的Rboh酶的特异性抑制剂)相关的显色底物NBT,结合共聚焦显微镜和体视显微镜,分别鉴定了一般ROS的细胞定位和橄榄生殖组织中超氧化物的积累。活性氧和超氧化物的大量产生已经被描述,显示了在橄榄花发育过程中相当精确的空间和时间位置。添加DPI后NBT信号的降低表明,超氧化物的产生主要是由于Rboh或其他黄素氧化酶的活性。在亚细胞水平上,O2●−的积累位于成熟花粉和萌发花粉的质膜、粗内质网和线粒体中。
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引用次数: 0
Pulsed Electric Field-Based Extraction of Total Polyphenols from Sideritis raiseri Using Hydroethanolic Mixtures 基于脉冲电场的氢乙醇混合物提取葡萄干黄豆中总多酚的研究
Pub Date : 2022-04-19 DOI: 10.3390/oxygen2020008
V. Athanasiadis, Vasileios M. Pappas, D. Palaiogiannis, Theodoros G. Chatzimitakos, Eleni Bozinou, D. Makris, S. Lalas
Polyphenols are an important class of compounds, due to their excellent antioxidant properties. Lately, much effort has been placed into developing new extraction techniques and optimizing them, so that polyphenols can be retrieved more efficiently from the plant materials. One of the most recent advances in extraction techniques is pulsed electric field extraction (PEF). This new technique is environmentally friendly and has the potential to maximize the recovery of compounds from plant tissues. Although the efficiency of PEF depends, among others, on the nature of the solvent used, up to date, there are no reports on the optimization of the PEF extraction of polyphenols, using hydroethanolic solutions of varying content in ethanol. In this study, three hydroethanolic solutions, water, and ethanol were used for the PEF-based extraction of total polyphenols from Sideritis raiseri. Results were conclusive that the 1:1 mixture of ethanol and water can increase by up to 146% the yield of polyphenols in the extract, highlighting the need to study more extensively, in the future, mixtures of solvents and not just plain water.
多酚是一类重要的化合物,具有优异的抗氧化性能。近年来,人们一直在努力开发新的提取技术并对其进行优化,以便更有效地从植物材料中提取多酚。脉冲电场萃取(PEF)是萃取技术的最新进展之一。这项新技术是环保的,有可能最大限度地从植物组织中回收化合物。虽然PEF的效率取决于所用溶剂的性质,但迄今为止,还没有关于PEF提取多酚的优化报道,使用不同含量的乙醇氢乙醇溶液。在本研究中,采用三种乙醇溶液,水和乙醇,以pef为基础提取葡萄干黄芪中的总多酚。结果是决定性的,乙醇和水的1:1混合物可以增加提取物中多酚的产量高达146%,强调需要更广泛地研究,在未来,溶剂的混合物,而不仅仅是普通的水。
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引用次数: 14
Free Radical Properties, Source and Targets, Antioxidant Consumption and Health 自由基的性质,来源和目标,抗氧化剂的消耗和健康
Pub Date : 2022-04-12 DOI: 10.3390/oxygen2020006
G. Martemucci, C. Costagliola, M. Mariano, L. D’andrea, P. Napolitano, A. D’Alessandro
Free radicals have acquired growing importance in the fields of biology and medicine. They are produced during many different endogenous and exogenous processes. Mitochondria are the main source of endogenous reactive oxygen species (ROS) produced at cell level. The overproduction of free radicals can damage macromolecules such as nucleic acids, proteins and lipids. This leads to tissue damage in various chronic and degenerative diseases. Antioxidants play a crucial role in the body’s defense against free radicals. This review concerns the main properties of free radicals, their sources and deleterious effects. It highlights the potential role of the dietary supplementation of antioxidants and discusses unsolved problems regarding antioxidant supplements in the prevention and therapy of diseases.
自由基在生物学和医学领域的重要性与日俱增。它们在许多不同的内源性和外源性过程中产生。线粒体是细胞水平产生内源性活性氧(ROS)的主要来源。自由基的过量产生会破坏大分子,如核酸、蛋白质和脂质。这导致各种慢性和退行性疾病的组织损伤。抗氧化剂在人体抵御自由基方面起着至关重要的作用。本文综述了自由基的主要性质、来源和有害作用。它强调了膳食补充抗氧化剂的潜在作用,并讨论了抗氧化剂补充剂在预防和治疗疾病方面尚未解决的问题。
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引用次数: 123
Lung Diffusing Capacity in Dutch Special Operations Forces Divers Exposed to Oxygen Rebreathers over 18 Years 荷兰特种作战部队潜水员暴露于氧气再呼吸装置18年的肺扩散能力
Pub Date : 2022-03-31 DOI: 10.3390/oxygen2020005
Tom H.B. den Ouden, T. Wingelaar, E. Endert, P. V. van Ooij
Exposure to hyperoxic conditions can induce pulmonary oxygen toxicity (POT). Divers of the Special Operations Forces (SOF) use oxygen rebreather systems during dives, and therefore are frequently exposed to hyperoxic conditions. Few studies have reported on POT in this population. This study reports on long-term pulmonary function tests (PFTs) and diffusing capacity in SOF divers to test the hypothesis that these measures of pulmonary function do not change clinically significantly during their career. The Royal Netherlands Navy performs yearly medical assessments of its military divers. All PFT and diffusing capacity data of SOF divers between the years 2000 and 2020 were analyzed using generalized estimating equations. The study included 257 SOF divers (median age, 27; interquartile range, 24–32), with 1612 dive medical assessments and a maximum follow-up time of 18.8 years. Alveolar volume (VA) and the diffusing capacity of carbon monoxide (TLCO) were significantly lower at baseline in smokers. Although these parameters were within the normal range, they declined over time and were significantly associated with age and years of diving. Smoking additionally affected TLCO and the transfer coefficient for carbon monoxide (KCO). TLCO and KCO were reduced by years of diving with oxygen rebreathers, albeit over clinically insignificant ranges, but smoking increased these changes by factors of 10 and 15, respectively.
暴露在高氧条件下可诱发肺氧毒性(POT)。特种作战部队(SOF)的潜水员在潜水过程中使用氧气再呼吸系统,因此经常暴露在高氧条件下。很少有研究报道这种人群中的POT。本研究报告了SOF潜水员的长期肺功能测试(PFTs)和扩散能力,以检验这些肺功能测量在其职业生涯中不会发生临床显著变化的假设。荷兰皇家海军每年对其军事潜水员进行医疗评估。使用广义估计方程分析了2000年至2020年间SOF潜水员的所有PFT和扩散能力数据。该研究包括257名SOF潜水员(中位年龄为27岁;四分位间距为24-32岁),进行了1612次潜水医学评估,最长随访时间为18.8年。吸烟者的肺泡容积(VA)和一氧化碳扩散能力(TLCO)在基线时显著降低。尽管这些参数在正常范围内,但随着时间的推移,它们会下降,并且与年龄和潜水年限显著相关。吸烟还影响TLCO和一氧化碳(KCO)的传递系数。使用氧气呼吸器潜水多年后,TLCO和KCO降低,尽管在临床上不显著,但吸烟使这些变化分别增加了10和15倍。
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引用次数: 1
A Brief History of Oxygen: 250 Years on 《氧气简史:250年
Pub Date : 2022-03-15 DOI: 10.3390/oxygen2010004
J. Hancock
Although there has been some controversy surrounding exactly when oxygen was first discovered, it is likely that that accolade should go to Carl Wilhelm Scheele, who isolated oxygen in 1772, or even a year earlier. Others since then have been given the credit for the instrumental work leading to the discovery including Joseph Priestley in 1774 and Antoine-Laurent Lavoisier. Oxygen, a paramagnetic, diradical gaseous (at room temperature) molecule, is instrumental to life as we know it. It is also crucial to some medical therapies, used in multiple industries and has even been found on other planets. The importance of oxygen cannot be overplayed. Now, 250 years since oxygen was discovered, it is timely to revisit some of the history, the controversies and look at how oxygen has evolved during that time. Here, a few of the highlights in oxygen research are discussed.
尽管关于氧气首次被发现的确切时间存在一些争议,但这一荣誉很可能应该归于1772年甚至一年前分离出氧气的卡尔·威廉·谢尔。自那以后,其他人因其发现的工具性工作而受到赞誉,包括1774年的约瑟夫·普里斯特利和安托万·劳伦特·拉瓦锡。氧是一种顺磁性、自由基气态(室温下)分子,对我们所知的生命至关重要。它对一些医疗疗法也至关重要,用于多个行业,甚至在其他行星上也有发现。氧气的重要性再怎么强调也不为过。现在,氧气被发现已经250年了,现在是时候重温一些历史、争议,看看氧气在那段时间是如何进化的。在这里,讨论了氧气研究中的一些亮点。
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引用次数: 2
An Inexpensive Incubator for Mammalian Cell Culture Capable of Regulating O2, CO2, and Temperature 一种可调节O2、CO2和温度的廉价哺乳动物细胞培养箱
Pub Date : 2022-03-14 DOI: 10.3390/oxygen2010003
P. Samokhin, Georgina L. Gardner, Christopher Moffatt, J. Stuart
Mammalian cell culture is widely used for discovery and development. Recently, increasing attention has been paid to the importance of maintaining physiologically-relevant conditions in cell culture. Although oxygen level is a particularly important consideration, it is rarely regulated by experimentalists. The atmospheric O2 levels commonly used in cell culture are significantly higher than those experienced by most mammalian cells in vivo, leaving cells susceptible to oxidative damage, senescence, transformation, and otherwise aberrant physiology. A barrier to incorporating O2 regulation into most cell culture workflows has been the expense of investing in new equipment, as the vast majority of laboratory CO2 incubators do not regulate O2. Here, we describe an inexpensive (
哺乳动物细胞培养被广泛用于发现和开发。最近,在细胞培养中保持生理相关条件的重要性得到了越来越多的关注。尽管氧气水平是一个特别重要的考虑因素,但它很少受到实验者的调节。细胞培养中常用的大气O2水平显著高于大多数哺乳动物细胞在体内所经历的水平,使细胞容易受到氧化损伤、衰老、转化和其他异常生理学的影响。将O2调节纳入大多数细胞培养工作流程的一个障碍是投资新设备的费用,因为绝大多数实验室CO2培养箱都不调节O2。在这里,我们描述了一种价格低廉(<CAD 1000)、便携式且用户友好的O2/CO2培养箱,它可以在生理范围内建立和保持生理O2、CO2和温度值。我们使用了一种基于Arduino的方法,将O2和CO2控制添加到温度调节鸡蛋培养箱中。我们的培养箱在商业实验室O2/CO2培养箱中进行了测试。使用Presens-OxoDish技术,我们证明,在5%气相培养箱O2的设定值下,培养基O2平均为5.03(SD=0.03),范围为4.98-5.09%。在培养箱中培养的MCF7、LNCaP和C2C12细胞系显示出正常的形态、增殖和活力。长达一周的培养没有产生污染。因此,我们的培养箱提供了一种在常规哺乳动物细胞培养中维持生理机能的廉价方法。
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引用次数: 3
Prolonged Cold Ischemia Did Not Impair Mitochondrial Oxygen Consumption or Reactive Oxygen Species Production in Human Uterine Fundus and Horn Myometrium 长时间冷缺血不会影响人子宫底和角肌线粒体耗氧量或活性氧的产生
Pub Date : 2022-02-28 DOI: 10.3390/oxygen2010002
Mathilde Pélissié, A. Charles, F. Goupilleau, I. Georg, A. Bryand, B. Gény, O. Garbin
Uterine transplantation may be a solution for infertility of uterine origin. Nevertheless, only three pregnancies with a live birth have so far been possible involving a uterine transplant from a brain-dead donor. Particularly, the impact of ischemia needs a better understanding. Analysis of mitochondrial respiration and production of reactive oxygen species (ROS) in muscle are of interest since they are pertinent markers of the harmful effects of ischemia. We therefore studied both uterine fundus and horn muscle mitochondrial use of oxygen and ROS production in eight women needing hysterectomy. High resolution respirometry and electron paramagnetic resonance allowed the determination of, respectively, myometrium oxidative capacity, hydrogen peroxide, mitochondrial free radical leak and superoxide anion production early (2 and 7 h) and late (24 h) following surgery. Mitochondrial oxygen consumption of the uterine fundus and horn tended to decrease with time but this was not statistically significant. Concerning ROS production, globally, we observed no significant change for H2O2, superoxide anion and free radical leak. In conclusion, a long period of cold ischemia did not impair myometrium mitochondrial respiration, only generating a transient H2O2 increase in uterine fundus. These data support that cold ischemia, even when prolonged, does not significantly alter uterine muscle oxidative capacity.
子宫移植可能是子宫源性不孕症的一种解决方案。然而,到目前为止,只有三名孕妇通过脑死亡捐赠者的子宫移植而活产。特别是,缺血的影响需要更好的理解。线粒体呼吸和肌肉中活性氧(ROS)产生的分析是有意义的,因为它们是缺血有害影响的相关标记。因此,我们研究了8名需要子宫切除术的妇女的子宫底和角肌线粒体对氧气和活性氧的使用。高分辨率呼吸法和电子顺磁共振分别测定手术后早期(2和7小时)和晚期(24小时)肌层氧化能力、过氧化氢、线粒体自由基泄漏和超氧阴离子产生。子宫底和子宫角线粒体耗氧量随时间的延长而减少,但无统计学意义。在ROS生成方面,我们在全球范围内观察到H2O2、超氧阴离子和自由基泄漏没有明显变化。综上所述,长时间的冷缺血不影响子宫肌层线粒体呼吸,仅使子宫底过氧化氢短暂升高。这些数据支持冷缺血,即使延长,也不会显著改变子宫肌氧化能力。
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引用次数: 0
Beneficial Effects of Antioxidants in Male Infertility Management: A Narrative Review 抗氧化剂在男性不育症治疗中的有益作用:综述
Pub Date : 2022-01-28 DOI: 10.3390/oxygen2010001
S. Cilio, Monica Rienzo, Gianluca Villano, Benito Fabio Mirto, Gaetano Giampaglia, Federico Capone, Gianpiero Ferretti, E. di Zazzo, F. Crocetto
Background: Infertility, defined as the failure to conceive after one year of regular, unprotected intercourse, affects 50–80 million people worldwide. A male factor is involved in approximately 20–30% of cases. In the etiology of male infertility, the association between poor semen quality and oxidative stress (OS) is well known. High levels of reactive oxygen species (ROS) allow the oxidation of DNA, proteins, and lipids of sperm cells, modifying their vitality, motility, and morphology. Methods: To evaluate the effects of antioxidants on sperm in infertile men, we queried the MEDLINE database (via the PubMed interface) for published studies in the last 10 years (2011–2021). The following keywords were used: “infertility” and -“inositol”, -“alpha-lipoic acid”, -“zinc”, -“folate”, -“coenzyme Q10”, -“selenium”, and -“vitamin”. Results: Inositol regulates OS levels in sperm cells thanks to its role in mitochondrial reactions and is involved in several processes favoring sperm–oocyte interactions. Alpha-lipoic acid (ALA) reduces ROS damage and improves semen parameters in terms of spermatozoa’s motility, morphology, and count. Poor zinc nutrition may be related to low quality of sperm. Supplementation of folate plus zinc has a positive effect on the sperm concentration and morphology. Supplementation with CoQ10 increases sperm concentration, total and progressive motility. Selenium (Se) supplementation improves the overall semen quality and is related to a higher ejaculated volume. Among vitamins, only vitamin B12 shows a positive effect on semen quality; it increases sperm count and motility and reduces sperm DNA damage. Conclusions: In men showing low-quality semen, diet supplementation with antioxidants may improve the sperm quality by alleviating OS-induced sperm damage and enhancing hormone synthesis and spermatozoa concentration, motility, and morphology. Future clinical trials should be focused on the possible association of several antioxidants to take advantage of combined mechanisms of action.
背景:不孕症的定义是在一年的定期无保护性交后仍不能怀孕,全世界有5000万至8000万人受到影响。大约20-30%的病例涉及男性因素。在男性不育的病因学中,精液质量差与氧化应激(OS)之间的关系是众所周知的。高水平的活性氧(ROS)允许精子细胞的DNA、蛋白质和脂质氧化,改变它们的活力、运动性和形态。方法:为了评估抗氧化剂对不育男性精子的影响,我们查询了MEDLINE数据库(通过PubMed界面)最近10年(2011-2021)发表的研究。使用了以下关键词:“不育”和“肌醇”、“α -硫辛酸”、“锌”、“叶酸”、“辅酶Q10”、“硒”和“维生素”。结果:肌醇通过其在线粒体反应中的作用调节精子细胞中的OS水平,并参与一些有利于精子-卵母细胞相互作用的过程。α -硫辛酸(ALA)减少活性氧损伤,改善精子活力、形态和数量方面的精液参数。锌营养不良可能与精子质量低有关。补充叶酸加锌对精子浓度和形态有积极影响。补充辅酶q10可提高精子浓度、总活力和进行性活力。硒(Se)的补充提高了精液的整体质量,并与更高的射精量有关。在维生素中,只有维生素B12对精液质量有积极影响;它能增加精子数量和活力,减少精子DNA损伤。结论:在精液质量较低的男性中,饮食中添加抗氧化剂可以通过减轻os引起的精子损伤、增强激素合成、精子浓度、活力和形态来改善精子质量。未来的临床试验应侧重于几种抗氧化剂的可能关联,以利用联合作用机制。
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引用次数: 11
Role of Oxygen Radicals in Alzheimer’s Disease: Focus on Tau Protein 氧自由基在阿尔茨海默病中的作用:以Tau蛋白为重点
Pub Date : 2021-11-27 DOI: 10.3390/oxygen1020010
A. Atlante, D. Valenti, V. Latina, G. Amadoro
Oxygen free radical burst is a prominent early event in the pathogenesis of Alzheimer’s disease (AD). Posttranslational modifications of Tau protein, primarily hyper-phosphorylation and truncation, are indicated as critical mediators of AD pathology. This finding is confirmed by the high levels of oxidative stress markers and by the increased susceptibility to oxygen radicals found in cultured neurons and in brains from transgenic animal models expressing toxic Tau forms, in concomitance with a dramatic reduction in their viability/survival. Here, we collect the latest progress in research focused on the reciprocal and dynamic interplay between oxygen radicals and pathological Tau, discussing how these harmful species cooperate and/or synergize in the progression of AD. In this context, a better understanding of the role of oxidative stress in determining Tau pathology, and vice versa, primarily could be able to define novel biomarkers of early stages of human tauopathies, including AD, and then to develop therapeutic strategies aimed at attenuating, halting, or reversing disease progression.
氧自由基爆发是阿尔茨海默病(AD)发病的重要早期事件。Tau蛋白的翻译后修饰,主要是超磷酸化和截断,被认为是AD病理的关键介质。这一发现被高水平的氧化应激标记物和在表达有毒Tau形式的转基因动物模型中培养的神经元和大脑中发现的对氧自由基的易感性增加所证实,同时它们的生存能力/存活率急剧降低。在这里,我们收集了关于氧自由基与病理Tau之间相互作用和动态相互作用的最新研究进展,讨论了这些有害物种在AD进展中的合作和/或协同作用。在这种情况下,更好地理解氧化应激在决定Tau病理中的作用,反之亦然,主要能够定义包括AD在内的人类Tau病变早期阶段的新生物标志物,然后制定旨在减轻,停止或逆转疾病进展的治疗策略。
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引用次数: 4
The Reactive Oxygen Species Singlet Oxygen, Hydroxy Radicals, and the Superoxide Radical Anion—Examples of Their Roles in Biology and Medicine 活性氧单线态氧、羟基自由基和超氧自由基阴离子——它们在生物学和医学中的作用的例子
Pub Date : 2021-10-27 DOI: 10.3390/oxygen1020009
R. Edge, T. Truscott
Reactive oxygen species comprise oxygen-based free radicals and non-radical species such as peroxynitrite and electronically excited (singlet) oxygen. These reactive species often have short lifetimes, and much of our understanding of their formation and reactivity in biological and especially medical environments has come from complimentary fast reaction methods involving pulsed lasers and high-energy radiation techniques. These and related methods, such as EPR, are discussed with particular reference to singlet oxygen, hydroxy radicals, the superoxide radical anion, and their roles in medical aspects, such as cancer, vision and skin disorders, and especially pro- and anti-oxidative processes.
活性氧物种包括基于氧的自由基和非自由基物种,例如过氧亚硝酸盐和电子激发(单线态)氧。这些反应性物种的寿命通常很短,我们对它们在生物环境,特别是医疗环境中的形成和反应性的大部分理解都来自于脉冲激光和高能辐射技术的互补快速反应方法。这些和相关的方法,如EPR,特别是单重态氧、羟基自由基、超氧化物自由基阴离子及其在医学方面的作用进行了讨论,如癌症、视力和皮肤疾病,尤其是促氧化和抗氧化过程。
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引用次数: 12
期刊
Oxygen (Basel, Switzerland)
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