Abstract:
:The use of antioxidants to prevent intracellular free radical damage is an area currently attracting considerable research interest. The compound 2',7'-dichlorofluorescin diacetate (DCFH-DA) is a probe for intracellular peroxide formation commonly used in such studies. During our studies we unexpectedly found that incubation of Trolox, a water soluble vitamin E analog, with DCFH-DA in cell-free physiological buffers resulted in the deacetylation and oxidation of DCFH-DA to form the fluorescent compound, 2',7'-dichlorofluorescein (DCF). The reaction was time-, temperature-, and pH-dependent. Fluorescence intensity increased with an increase in either Trolox or DCFH-DA concentration. These results indicate that even at physiological pH, DCFH-DA can be deacetylated to form 2',7'-dichlorofluororescin (DCFH), DCFH can then be oxidized to DCF by abstraction of a hydrogen atom by the phenoxyl radical of Trolox. Exposure of the reaction mixture to 10 Gy of 60Co gamma radiation greatly increased production of DCF. Antioxidant compounds reported to "repair" the Trolox phenoxyl radical (e.g., ascorbic acid, salicylate) can also prevent the Trolox-induced DCFH-DA fluorescence. However, radical (e.g., catechin) or can themselves form a radical (e.g., uric acid, TEMPOL) either have no effect or can increase levels of DCF. These results demonstrate that experimental design must be carefully considered when using DCFH-DA to measure peroxide formation in combination with certain antioxidants.
journal_name
Free Radic Resjournal_title
Free radical researchauthors
Kalinich JF,Ramakrishnan N,McClain DEdoi
10.3109/10715769709097782subject
Has Abstractpub_date
1997-01-01 00:00:00pages
37-47issue
1eissn
1071-5762issn
1029-2470journal_volume
26pub_type
杂志文章abstract::The ubiquitin-proteasome system (UPS) and autophagy are two major intracellular proteolytic systems that are closely associated with each other. Because UPS and autophagy are involved in the clearance of oxidised and/or aggregated proteins, it would be logical to assume that alterations in proteolysis would accompany ...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.1080/10715762.2020.1734588
更新日期:2020-04-20 00:00:00
abstract::Oxygen radical generating systems, namely, Cu(II)/ H2O2, Cu(II)/ascorbate, Cu(II)/NAD(P)H, Cu(II)/ H2O2/catecholamine and Cu(II)/H2O2/SH-compounds irreversibly inhibited yeast glutathione reductase (GR) but Cu(II)/H2O2 enhanced the enzyme diaphorase activity. The time course of GR inactivation by Cu(II)/H2O2 dependent...
journal_title:Free radical research
pub_type: 杂志文章,评审
doi:10.3109/10715769709097858
更新日期:1997-12-01 00:00:00
abstract::We have determined the effects of chronic vitamin C intake on neutrophil and lymphocyte antioxidant defences during the acute phase immune response induced by intense exercise. Blood samples were taken from 16 voluntary athletes in basal conditions, both immediately after and 1 h after a duathlon competition. Sportsme...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.1080/1071576031000150454
更新日期:2003-09-01 00:00:00
abstract::The intestinal tract, known for its capability for self-renew, represents the first barrier of defence between the organism and its luminal environment. The thiol/disulfide redox systems comprising the glutathione/glutathione disulfide (GSH/GSSG), cysteine/cystine (Cys/CySS) and reduced and oxidized thioredoxin (Trx/T...
journal_title:Free radical research
pub_type: 杂志文章,评审
doi:10.3109/10715762.2011.611509
更新日期:2011-11-01 00:00:00
abstract::Iron or oxygen regulates the stability of hypoxia inducible factor-1α (HIF-1α). We investigated whether ferrous glycinate would affect HIF-1α accumulation, aerobic glycolysis and mitochondrial energy metabolism in human A549 lung cancer cells. Incubation of A549 cells with ferrous glycinate decreased the protein level...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.1080/10715762.2018.1476691
更新日期:2018-12-01 00:00:00
abstract::The imbalance between the reactive oxygen (ROS) and nitrogen (RNS) species production and their handling by the antioxidant machinery (low molecular weight antioxidant molecules and antioxidant enzymes), also known as oxidative stress, is a condition caused by physiological and pathological processes. Moreover, oxidat...
journal_title:Free radical research
pub_type: 杂志文章,评审
doi:10.1080/10715762.2018.1542141
更新日期:2019-02-01 00:00:00
abstract:PURPOSE:Aspiration pneumonia is infection of the respiratory tract resulting from accumulation of sputum in the larynx. N-acetyl-L-cysteine (NAC) might regulate mucin (MUC) expression and activate inherent anti-infective system in bronchiolar epithelial cells after cellular uptake, and therefore, serve as the preventat...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.3109/10715762.2015.1087642
更新日期:2015-01-01 00:00:00
abstract::We tested the hypothesis that hypertension associated with polycythemia vera (PV) may be related to hemoglobin released from erythrocytes (cell-free hemoglobin, fHb). We assessed hematocrit, mean arterial pressure (MAP), blood viscosity, and the level of fHb and nitrite/nitrate (NOx) in the plasma of 73 PV patients an...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.3109/10715762.2013.860225
更新日期:2014-02-01 00:00:00
abstract::It is well known that reactive oxygen species (ROS) attack several living tissues and increase the risk of development and progression of serious diseases. In neuronal level, ROS induce cell death in concentration-dependent fashion. However, little is known about the mechanisms of neuronal changes by ROS prior to indu...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.3109/10715762.2011.647689
更新日期:2012-02-01 00:00:00
abstract::Membrane rafts (MRs) are specialized microdomains in the cell membrane with an altered lipid composition. Upon various stimulations, MRs can be clustered to aggregate or recruit NADPH oxidase sub-units and related proteins to form MR redox signalosomes in the membrane of cells like vascular endothelial cells (ECs). Mu...
journal_title:Free radical research
pub_type: 杂志文章,评审
doi:10.3109/10715762.2010.485994
更新日期:2010-08-01 00:00:00
abstract::Tumour necrosis factor alpha (TNF alpha) cytotoxicity is mediated, at least in part, by oxidative stress and phospholipase A2 activation. The first post-receptor events to be observed in TNF alpha-sensitive lines are the generation of superoxide anion (O2-) within the mitochondria and the activation of phospholipase A...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.3109/10715769609149917
更新日期:1996-08-01 00:00:00
abstract::The aim of the study was to investigate whether there is transmembrane transport of intact glutathione ([3H]-GSH, 0.1 microCi) in rat and human type II pneumocytes (T2P), and if this transport might be dependent on the redox state of the extracellular fluid. The T2P were pretreated with acivicin (250 microM) to inhibi...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.1080/10715769900300411
更新日期:1999-05-01 00:00:00
abstract::A growing body of experimental evidence suggests that the oxidative neurotoxicity of hemoglobin A may contribute to neuronal loss after CNS hemorrhage. Several hemoglobin variants, including hemoglobin S, are more potent oxidants in cell-free systems. However, despite the increased incidence of hemorrhagic stroke asso...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.1080/10715760310001638010
更新日期:2004-05-01 00:00:00
abstract::MCI-186 (3-methyl-1-phenyl-2-pyrazolin-5-one) is a newly developed antioxidant which has been shown to reduce brain edema in cerebral ischemia through inhibition of the lipoxygenase pathway of arachidonic acid. However, its effect on myocardial reperfusion injury after prolonged ischemia has not yet been demonstrated....
journal_title:Free radical research
pub_type: 杂志文章
doi:10.3109/10715769609088034
更新日期:1996-05-01 00:00:00
abstract::To solve the problems faced in clinical use of hypocrellins, a water-soluble preparation of Hypocrellin B (HB), HB-Triton X-100 (TX-100) micelles, was prepared. To evaluate the photodynamic activity, the free radicals (*OH and HB*-) and singlet oxygen (1O2) generated via photosensitization of the preparation in aqueou...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.1080/10715760310001601056
更新日期:2003-10-01 00:00:00
abstract::7,8-Dihydroneopterin protects cells intracellularly from oxidative stress-induced death, but its mode of transport across the cell membrane is unknown. Nucleosides, such as guanosine, are transported via nucleoside transporters of the equilibrative and concentrative forms. Therefore, the objective of this study was to...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.1080/10715762.2020.1764948
更新日期:2020-05-01 00:00:00
abstract::Degenerative diseases such as cancer are induced by oxidative genetic damage. Antioxidants can scavenge reactive oxygen species, but to prevent disease, they must do so quickly, before the DNA bases are damaged. In the present study, a novel method was established for evaluating the potency of antioxidants employing 2...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.1080/10715760290019336
更新日期:2002-03-01 00:00:00
abstract::Macrophages produce reactive oxygen species such as O2-, H2O2 and *OH that contribute to the pathogenesis of diseases such as inflammation and atherosclerosis. The cells have multiple defense systems against those reactive oxygen species, and we describe here such an oxidative stress-inducible defense system. Upon exp...
journal_title:Free radical research
pub_type: 杂志文章,评审
doi:10.1080/10715769900300921
更新日期:1999-10-01 00:00:00
abstract::Liposomes composed of soybean phosphatidylcholine were peroxidized using the reagent sodium hypochlorite or the myeloperoxidase-hydrogen peroxide-Cl- system. Linoleic acid hydroperoxide previously prepared from linoleic acid by means of lipoxidase was incorporated into liposomes. The yield of thiobarbituric acid react...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.1080/10715769900300521
更新日期:1999-06-01 00:00:00
abstract::APE1/Ref-1, the mammalian ortholog of E. coli Xth, and a multifunctional protein possessing both DNA repair and transcriptional regulatory activities, has dual role in controlling cellular response to oxidative stress. It is rate-limiting in repair of oxidative DNA damage including strand breaks and also has co-transc...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.1080/10715760701765616
更新日期:2008-01-01 00:00:00
abstract::Human serum samples were evaluated for oxidative stress with the d-ROMs test. The ceruloplasmin (CP) and copper contents of the samples was independently measured and compared to those calculated on the basis of the d-ROMs test results for pure CP solutions. The d-ROMs readings did not show any significant correlation...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.3109/10715762.2015.1136063
更新日期:2016-01-01 00:00:00
abstract::The use of salicylate as a chemical trap for .OH represents a simple and convenient alternative to the use of spin trapping techniques to study oxidative injury in isolated perfused organs. In these systems, salicylate is included in the perfusion buffer at concentrations ranging from 0.1 to 2 mM depending on the dete...
journal_title:Free radical research
pub_type: 杂志文章,评审
doi:10.3109/10715769409056588
更新日期:1994-11-01 00:00:00
abstract::The effects of inorganic phosphate (Pi), the main intracellular membrane permeable anion capable of altering mitochondrial pH gradients (Delta pH), were measured on mitochondrial H2O2 release. As expected, Pi decreased Delta pH and increased the electric membrane potential (Delta Psi). Mitochondrial H2O2 release was s...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.1080/10715760400009258
更新日期:2004-10-01 00:00:00
abstract::The hydroxyl radical (.OH)-scavenging activity of d-2-[4-(3-methyl-2-thienyl)phenyl]propionic acid (M-5011), a novel nonsteroidal anti-inflammatory drug (NSAID), and that of several other NSAIDs were investigated by the hyaluronic acid (HA) degradation method and the electron spin resonance (ESR) spin-trapping techniq...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.3109/10715769709065781
更新日期:1997-10-01 00:00:00
abstract::Prostaglandin E2 (PGE2) has been reported to play critical roles in cell fate decision by interacting with four types of prostanoid receptors such as EP1, EP2, EP3 and EP4. The present study was aimed at investigating the effect of the EP4-specific agonist CAY10598 in human colon cancer HCT116 cells. Our study reveale...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.3109/10715762.2014.951838
更新日期:2014-11-01 00:00:00
abstract::Antioxidants are important species which possess the ability to protect the body from damage caused by free radical-induced oxidative stress. A variety of free radical-scavenging antioxidants exist within the body many of which are derived from dietary sources. There is currently much interest in the antioxidant role ...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.3109/10715769709097807
更新日期:1997-03-01 00:00:00
abstract::We have here investigated possible occurrence of bicarbonate-dependent, carbonate radical anion (CO(3)(•-))-driven tocopherol-mediated human LDL peroxidation (TMP) in vitro and in vivo. CO(3)(•-), generated in vitro by the SOD1/H(2)O(2)/bicarbonate system, readily promoted TMP, which was dependent on α-tocopherol and ...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.3109/10715762.2012.719613
更新日期:2012-11-01 00:00:00
abstract::The hydroxyl radical adducts of 5,5 dimethyl-1-pyrolline-N-oxide (DMPO) and 3,3,5,5 tetramethyl-1-pyrolline-N-oxide (TMPO) formed in the presence of hydrogen peroxide and FeII are normally quite stable, but in the presence of 5-20 micromolar myoglobin their ESR signals decay rapidly. This decay probably reflects furth...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.3109/10715769409145632
更新日期:1994-05-01 00:00:00
abstract::Reactive oxygen species (ROS) are involved in the mechanism of photoaging and carcinogenesis. Skin is endowed with antioxidant enzymes including superoxide dismutases (SOD): cytosolic copper zinc SOD and mitochondrial manganese SOD. The aim of our study was to estimate the protective effect of manganese against oxidat...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.3109/10715769509065255
更新日期:1995-10-01 00:00:00
abstract::Thioredoxin-1 (TRX) is a redox-active protein with anti-inflammatory effects. This study investigated the optimal delivery method and the mechanisms of recombinant human TRX (rhTRX) to suppress neutrophil recruitment in a rat bleomycin (BLM)-induced sustained acute lung injury model. In male Wister rats intratracheall...
journal_title:Free radical research
pub_type: 杂志文章
doi:10.1080/10715760701487682
更新日期:2007-10-01 00:00:00