Ischemia-reperfusion injury induces cell death, but the responsible mechanisms are not understood. This study examined mitochondrial depolarization and cell death during ischemia and reperfusion. Contracting cardiomyocytes were subjected to 60-min ischemia followed by 3-h reperfusion. Mitochondrial membrane potential (DeltaPsi(m)) was assessed with tetramethylrhodamine methyl ester. During ischemia, DeltaPsi(m) decreased to ... More
Improved efficacy of stem cell labeling for magnetic resonance imaging studies by the use of cationic liposomes.
Authorsvan den Bos EJ, Wagner A, Mahrholdt H, Thompson RB, Morimoto Y, Sutton BS, Judd RM, Taylor DA
JournalCell Transplant
PubMed ID14653621
'Labeling stem cells with FDA-approved superparamagnetic iron oxide particles makes it possible to track cells in vivo with magnetic resonance imaging (MRI), but high intracellular levels of iron can cause free radical formation and cytotoxicity. We hypothesized that the use of cationic liposomes would increase labeling efficiency without toxic effects. ... More
Action of DCFH and BODIPY as a probe for radical oxidation in hydrophilic and lipophilic domain.
AuthorsYoshida Y, Shimakawa S, Itoh N, Niki E
JournalFree Radic Res
PubMed ID14567446
'Fluorogenic probes such as 2'',7''-dichlorofluorescin (DCFH) have been extensively used to detect oxidative events and to measure antioxidant capacity. At the same time, however, the inherent drawbacks of these probes such as non-specificity towards oxidizing species have been pointed out. The present study was carried out to analyze the action ... More
A method to measure the oxidizability of both the aqueous and lipid compartments of plasma.
AuthorsAldini G, Yeum KJ, Russell RM, Krinsky NI
JournalFree Radic Biol Med
PubMed ID11677037
'The lipophilic radical initiator (MeO-AMVN) and the fluorescent probe C11BODIPY581/591 (BODIPY) were used to measure the lipid compartment oxidizability of human plasma. Aqueous plasma oxidizability was initiated by the aqueous peroxyl radical generator, AAPH, and 2'',7''-dichlorodihydrofluorescein (DCFH) was employed as the marker of the oxidative reaction. The distribution in aqueous ... More
Detection of lipid peroxidation in equine spermatozoa based upon the lipophilic fluorescent dye C1l-BODIPY581/591.
AuthorsBall BA, Vo A
JournalJ Androl
PubMed ID11868820
'The lipophilic fluorescent probe, 4,4-difluoro-5-(4-phenyl-1 ,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid (C11-BODIPY581/591) was used to evaluate changes in lipid peroxidation in equine spermatozoa during both short-term exposure to ferrous sulfate and sodium ascorbate in the presence of cumene hydroperoxide as well as during storage of spermatozoa at 5 degrees C for 48 hours. Peroxidation ... More
(-)-Epigallocatechin-3-gallate prevents oxidative damage in both the aqueous and lipid compartments of human plasma.
AuthorsAldini G, Yeum KJ, Carini M, Krinsky NI, Russell RM
JournalBiochem Biophys Res Commun
PubMed ID12604363
'When human plasma was exposed to the hydrophilic radical initiator, AAPH, (-)-epigallocatechin-(3)-gallate (EGCG) dose-dependently inhibited the aqueous compartment oxidation (IC(50)=0.72 microM) (monitored by DCFH oxidation) and spared the lipophilic antioxidants, alpha-tocopherol, and carotenoids, but not ascorbic acid. When radicals were selectively induced in the lipid compartment by the lipophilic radical ... More
Mass spectrometric characterization of the oxidation of the fluorescent lipid peroxidation reporter molecule C11-BODIPY(581/591).
AuthorsDrummen GP, Gadella BM, Post JA, Brouwers JF
JournalFree Radic Biol Med
PubMed ID15182863
'C11-BODIPY(581/591) is a fluorescent lipid peroxidation reporter molecule that shifts its fluorescence from red to green when challenged with oxidizing agents, i.e., reactive oxygen species (ROS) or reactive nitrogen species (RNS). To understand the molecular mechanism responsible for this shift, we studied the molecular rearrangements leading to the shift in ... More
Fluorescence probes used for detection of reactive oxygen species.
AuthorsGomes A, Fernandes E, Lima JL,
JournalJ Biochem Biophys Methods
PubMed ID16297980
'Endogenously produced pro-oxidant reactive species are essential to life, being involved in several biological functions. However, when overproduced (e.g. due to exogenous stimulation), or when the levels of antioxidants become severely depleted, these reactive species become highly harmful, causing oxidative stress through the oxidation of biomolecules, leading to cellular damage ... More
Oxidation of unsaturated phospholipids in membrane bilayer mixtures is accompanied by membrane fluidity changes.
AuthorsBorst JW, Visser NV, Kouptsova O, Visser AJ
JournalBiochim Biophys Acta
PubMed ID10962288
'Steady-state and time-resolved fluorescence spectroscopy has been used to obtain information on oxidation processes and associated dynamical and structural changes in model membrane bilayers made from single unilamellar vesicles (SUV''s) of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) mixed with increasing amounts of 1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (SAPC). The highly unsaturated arachidonoyl chain containing four double bonds is ... More
Significance of mitochondrial reactive oxygen species in the generation of oxidative stress in spermatozoa.
'CONTEXT: Male infertility has been linked with the excessive generation of reactive oxygen species (ROS) by defective spermatozoa. However, the subcellular origins of this activity are unclear. OBJECTIVE: The objective of this study was to determine the importance of sperm mitochondria in creating the oxidative stress associated with defective sperm ... More
Fluorescent fatty acid to monitor reactive oxygen in single cells.
AuthorsPap EH, Drummen GP, Post JA, Rijken PJ, Wirtz KW
JournalMethods Enzymol
PubMed ID10907547
Amphiphilic, tri-block copolymers provide potent membrane-targeted neuroprotection.
AuthorsMarks JD, Pan CY, Bushell T, Cromie W, Lee RC
JournalFASEB J
PubMed ID11292683
Measuring mitochondrial reactive oxygen species.
AuthorsDegli Esposti M
JournalMethods
PubMed ID12054924
This article examines recent methods for measuring reactive oxygen species produced in isolated mitochondria and within live cells, with particular emphasis on the detection of hydrogen peroxide. Protocols for reliable measurements of mitochondrial hydrogen peroxide are presented, while the advantages and pitfalls of these and other methods are discussed. New ... More
Ratio-fluorescence microscopy of lipid oxidation in living cells using C11-BODIPY(581/591).
AuthorsPap EH, Drummen GP, Winter VJ, Kooij TW, Rijken P, Wirtz KW, Op den Kamp JA, Hage WJ, Post JA
JournalFEBS Lett
PubMed ID10405160
A ratio-fluorescence assay was developed for on-line localization and quantification of lipid oxidation in living cells. The assay explores the oxidative sensitivity of C11-BODIPY(581/591). Upon oxidation, the fluorescence of this fluorophore shifts from red to green. The probe incorporates readily into cellular membranes and is about twice as sensitive to ... More
Mass spectrometric analysis demonstrates that BODIPY 581/591 C11 overestimates and inhibits oxidative lipid damage.
AuthorsMacDonald ML, Murray IV, Axelsen PH,
JournalFree Radic Biol Med
PubMed ID17395012
BODIPY C11 is being used with increasing frequency to quantify lipid oxidation; however, it is not known whether signals from the dye yield accurate information. To determine the quantitative accuracy of signals from this dye we utilized a triple-quadrupole mass spectroscopy method to measure concentrations of 1-stearoyl-2-arachidonyl-sn-glycero-3-phosphocholine (SAPC) as it ... More
Electroporating fields target oxidatively damaged areas in the cell membrane.
Reversible electropermeabilization (electroporation) is widely used to facilitate the introduction of genetic material and pharmaceutical agents into living cells. Although considerable knowledge has been gained from the study of real and simulated model membranes in electric fields, efforts to optimize electroporation protocols are limited by a lack of detailed understanding ... More
Coordinated behavior of mitochondria in both space and time: a reactive oxygen species-activated wave of mitochondrial depolarization.
Reactive oxygen species (ROS) can trigger a transient burst of mitochondrial ROS production via ROS activation of the mitochondrial permeability transition pore (MPTP), a phenomenon termed ROS-induced ROS release (RIRR). The goal of this study was to investigate if the generation of ROS in a discrete region of a cardiomyocyte ... More
Regulation of sterol carrier protein gene expression by the forkhead transcription factor FOXO3a.
The SCP gene encodes two proteins, sterol carrier protein X (SCPx) and SCP2, that are independently regulated by separate promoters. SCPx has been shown to be the thiolase involved in the breakdown of branched-chain fatty acids and in the biosynthesis of bile acids. The in vivo function of SCP2 however ... More
Alpha-synuclein can function as an antioxidant preventing oxidation of unsaturated lipid in vesicles.
AuthorsZhu M, Qin ZJ, Hu D, Munishkina LA, Fink AL
JournalBiochemistry
PubMed ID16800638
Alpha-synuclein, a presynaptic protein associated with Parkinson's disease, is found as both soluble cytosolic and membrane-bound forms. Although the function of alpha-synuclein is unknown, several observations suggest that its association with membranes is important. In the present study we investigated the effect of alpha-synuclein on lipid oxidation in membranes containing ... More
Advantages and limitation of BODIPY as a probe for the evaluation of lipid peroxidation and its inhibition by antioxidants in plasma.
AuthorsItoh N, Cao J, Chen ZH, Yoshida Y, Niki E
JournalBioorg Med Chem Lett
PubMed ID17300936
Oxidative stress and the role of antioxidants are currently one of the most important subjects in the field of life science. In the present study, we assessed the oxidation of plasma lipids induced by free radicals and its inhibition by antioxidants with a fluorescence probe BODIPY. Vitamin E and C-depleted ... More
In vivo fluorescence correlation microscopy (FCM) reveals accumulation and immobilization of Nod factors in root hair cell walls.
Fluorescence correlation microscopy (FCM) is a new single-molecule detection technique based on the confocal principle to quantify molecular diffusion and concentration of fluorescent molecules (particles) with sub-micron resolution. In this study, FCM is applied to examine the diffusional behaviour of fluorescent Nod factor analogues on living Vicia sativa root hairs. ... More
C11-BODIPY(581/591), an oxidation-sensitive fluorescent lipid peroxidation probe: (micro)spectroscopic characterization and validation of methodology.
AuthorsDrummen GP, van Liebergen LC, Op den Kamp JA, Post JA
JournalFree Radic Biol Med
PubMed ID12160930
C11-BODIPY(581/591) is a fluorescent radio-probe for indexing lipid peroxidation and antioxidant efficacy in model membrane systems and living cells, with excellent characteristics: (i) emission in the visible range of the electromagnetic spectrum, with good spectral separation of the nonoxidized (595 nm) and oxidized (520 nm) forms; (ii) has a high ... More
Nod factors integrate spontaneously in biomembranes and transfer rapidly between membranes and to root hairs, but transbilayer flip-flop does not occur.
AuthorsGoedhart J, Röhrig H, Hink MA, van Hoek A, Visser AJ, Bisseling T, Gadella TW
JournalBiochemistry
PubMed ID10451386
Three novel nodulation (Nod) factors were synthesized from chitotetraose and three structurally different fluorescent BODIPY-tagged fatty acids. With fluorescence spectroscopic and microscopic techniques, the following aspects were studied: whether these amphiphilic molecules insert in membranes, whether they transfer between different membranes, and whether they are able to transfer from a ... More
Production of reactive oxygen species in mitochondria of HeLa cells under oxidative stress.
Mitochondria can be a source of reactive oxygen species (ROS) and a target of oxidative damage during oxidative stress. In this connection, the effect of photodynamic treatment (PDT) with Mitotracker Red (MR) as a mitochondria-targeted photosensitizer has been studied in HeLa cells. It is shown that MR produces both singlet ... More
Visualizing tubular lipid peroxidation in intact renal tissue in hypertensive rats.
AuthorsBapat S, Post JA, Braam B, Goldschmeding R, Koomans HA, Verkleij AJ, Joles JA
JournalJ Am Soc Nephrol
PubMed ID12444219
An imbalance between production of reactive oxygen species (ROS) and antioxidant defense is involved in the pathogenesis of diverse chronic parenchymatous diseases. To identify the primary site of such increased oxidative stress, a lipophilic ROS-sensitive probe (C11-Bodipy 581/591) is introduced, which allows the visualization and quantification of oxidative injury using ... More
In situ detection and localization of lipid peroxidation in individual bovine sperm cells.
AuthorsBrouwers JF, Gadella BM
JournalFree Radic Biol Med
PubMed ID14642386
Reactive oxygen species (ROS) have been implicated in many pathologies, including sub- and infertility. Freeze/thawing of sperm samples is routinely performed in the cattle breeding industries in order to perform artificial insemination. This freeze/thaw procedure is known to induce ROS in sperm samples. Lipid peroxidation in fresh and frozen/thawed sperm ... More
Attenuation of lipid peroxidation by antioxidants in rat-1 fibroblasts: comparison of the lipid peroxidation reporter molecules cis-parinaric acid and C11-BODIPY(581/591) in a biological setting.
AuthorsDrummen GP, Makkinje M, Verkleij AJ, Op den Kamp JA, Post JA
JournalBiochim Biophys Acta
PubMed ID15164761
Lipid peroxidation is a major factor in the pathogenesis of many disease states. To detect the initial stages of lipid peroxidation or evaluate antioxidant efficacy, cis-parinaric acid (cis-PnA) has been successfully used and thoroughly validated. However, cis-PnA is not very well suited for medium throughput screening of antioxidants in living ... More
Total antioxidant performance: a validated fluorescence assay for the measurement of plasma oxidizability.
The antioxidant capacity of human plasma was determined by following the oxidation kinetics of the lipid-soluble fluorescent marker BODIPY using 2,2'-azobis(4-methoxy-2,4-dimethylvaleronitrile) (MeO-AMVN) as the lipophilic radical initiator. The results are expressed as a total antioxidant performance (TAP) value based on the inhibition of BODIPY oxidation, as determined by the appearance ... More
A fluorometric method for measurement of peroxyl radical scavenging activities of lipophilic antioxidants.
AuthorsNaguib YM
JournalAnal Biochem
PubMed ID9882405
A fluorometric method for evaluation of activities of lipophilic antioxidants toward peroxyl radicals in a lipophilic medium (octane:butyronitrile; 9:1, v/v) and dioleoylphosphatidyl choline (DOPC) liposomal suspension in Tris-HCl buffer (pH 7.4 at 40 degrees C) has been developed. This fluorometric method is based on the use of the fatty acid ... More
Antioxidant activities of astaxanthin and related carotenoids.
AuthorsNaguib YM
JournalJ Agric Food Chem
PubMed ID10775364
The antioxidant activities of astaxanthin and related carotenoids have been measured by employing a newly developed fluorometric assay. This assay is based on 4,4-difluoro-3,5-bis(4-phenyl-1, 3-butadienyl)-4-bora-3a,4a-diaza-s-indacene (BODIPY 665/676) as an indicator; 2,2'-azobis-2,4-dimethylvaleronitrile (AMVN) as a peroxyl radical generator; and 6-hydroxy-2,5,7, 8-tetramethylchroman-2-carboxylic acid (Trolox) as a calibrator in an organic and liposomal ... More
Lipid diffusion in sperm plasma membranes exposed to peroxidative injury from oxygen free radicals.
AuthorsChristova Y, James PS, Jones R
JournalMol Reprod Dev
PubMed ID15112331
Unsaturated lipids in sperm plasma membranes are very susceptible to peroxidation when exposed to reactive oxygen species (ROS). In this investigation we have incubated ram spermatozoa in the presence of two ROS generating systems, ascorbate/FeSO4 and potassium peroxychromate (K3CrO8), and examined their effects on membrane fluidity by measuring fluorescence recovery ... More
The activities of antioxidant nutrients in human plasma depend on the localization of attacking radical species.
AuthorsYeum KJ, Aldini G, Chung HY, Krinsky NI, Russell RM
JournalJ Nutr
PubMed ID12888659
The oxidation of endogenous antioxidant nutrients in human plasma was determined to examine their activities against free radicals generated in the aqueous and lipid compartments of plasma. Free radicals were induced at a constant rate in the aqueous compartment by the hydrophilic radical generator, 2,2'-azobis-(2-amidinopropane)dihydrochloride (AAPH; 10-20 mmol/L) and in ... More
Intracellular dissemination of peroxidative stress. Internalization, transport, and lethal targeting of a cholesterol hydroperoxide species by sterol carrier protein-2-overexpressing hepatoma cells.
Sterol carrier protein-2 (SCP-2) plays a crucial role in the trafficking and metabolism of cholesterol and other lipids in mammalian cells. Lipid hydroperoxides generated under oxidative stress conditions are relatively long-lived intermediates that damage cell membranes and play an important role in redox signaling. We hypothesized that SCP-2-facilitated translocation of ... More
Development and validation of oxygen radical absorbance capacity assay for lipophilic antioxidants using randomly methylated beta-cyclodextrin as the solubility enhancer.
AuthorsHuang D, Ou B, Hampsch-Woodill M, Flanagan JA, Deemer EK
JournalJ Agric Food Chem
PubMed ID11902917
We recently reported the improved oxygen radical absorbance capacity (ORAC) assay using fluorescein (FL) as the fluorescent probe. The current ORAC(FL) assay is limited in hydrophilic antioxidant due to the aqueous environment of the assay. Lipophilic antioxidants mainly include the vitamin E family and carotenoids, which play a critical role ... More
Vitamin E inhibits lipid peroxidation-induced adhesion molecule expression in endothelial cells and decreases soluble cell adhesion molecules in healthy subjects.
Authorsvan Dam B, van Hinsbergh VW, Stehouwer CD, Versteilen A, Dekker H, Buytenhek R, Princen HM, Schalkwijk CG
JournalCardiovasc Res
PubMed ID12566129
OBJECTIVE: In a combination of in vivo and in vitro studies, we investigated mechanisms via which alpha-tocopherol, a lipid soluble form of vitamin E, can directly affect endothelial activation as induced by H(2)O(2) and TNFalpha. METHODS: We measured effects of alpha-tocopherol on H(2)O(2)-induced lipid peroxidation as determined with a fluorescent ... More
Nitroxidation, nitration, and oxidation of a BODIPY fluorophore by RNOS and ROS.
AuthorsNicolescu AC, Li Q, Brown L, Thatcher GR
JournalNitric Oxide
PubMed ID16549377
BODIPY C11 581/591 (BODIPY11) represents a sensitive probe for quantification of relative antioxidant capacity. However, the mechanism of BODIPY11 fluorescence decay in the presence of reactive oxygen species (ROS) and reactive nitrogen oxide species (RNOS) requires clarification. Azo-initiators provide a continuous source of peroxyl radicals that in simple, aerobic, homogeneous, ... More
Quantum dot-induced cell death involves Fas upregulation and lipid peroxidation in human neuroblastoma cells.
AuthorsChoi AO, Cho SJ, Desbarats J, Lovric J, Maysinger D
JournalJ Nanobiotechnology
PubMed ID17295922
BACKGROUND: Neuroblastoma, a frequently occurring solid tumour in children, remains a therapeutic challenge as existing imaging tools are inadequate for proper and accurate diagnosis, resulting in treatment failures. Nanoparticles have recently been introduced to the field of cancer research and promise remarkable improvements in diagnostics, targeting and drug delivery. Among ... More
Impact of oxidative stress in aged mouse oocytes on calcium oscillations at fertilization.
AuthorsTakahashi T, Takahashi E, Igarashi H, Tezuka N, Kurachi H
JournalMol Reprod Dev
PubMed ID12950101
In vivo post-ovulatory aging of oocytes significantly affects the development of oocytes and embryos. Also, oocyte aging alters the regulation of the intracellular calcium concentration, thus affecting Ca(2+) oscillations in fertilized oocytes. Because reactive oxygen species (ROS) are known to significantly perturb Ca(2+) homeostasis mainly through direct effects on the ... More
BAP1 links metabolic regulation of ferroptosis to tumour suppression.
Authors
JournalNat Cell Biol
PubMed ID30202049
Ferroptosis occurs through an osmotic mechanism and propagates independently of cell rupture.
Authors
JournalNat Cell Biol
PubMed ID32868903
NFS1 undergoes positive selection in lung tumours and protects cells from ferroptosis.
Bone Marrow Mesenchymal Stem Cells Support Acute Myeloid Leukemia Bioenergetics and Enhance Antioxidant Defense and Escape from Chemotherapy.
Authors
JournalCell Metab
PubMed ID32966766
3D Culture Models with CRISPR Screens Reveal Hyperactive NRF2 as a Prerequisite for Spheroid Formation via Regulation of Proliferation and Ferroptosis.
Authors
JournalMol Cell
PubMed ID33128871
PLCγ1 suppression promotes the adaptation of KRAS-mutant lung adenocarcinomas to hypoxia.
Authors
JournalNat Cell Biol
PubMed ID33077911
Cancer cells dying from ferroptosis impede dendritic cell-mediated anti-tumor immunity.
Authors
JournalNat Commun
PubMed ID35760796
A Compendium of Genetic Modifiers of Mitochondrial Dysfunction Reveals Intra-organelle Buffering.