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dihydrorhodamine 123 oxidation test

Michalina Grzesik, Katarzyna Naparło, Grzegorz Bartosz, Izabela Sadowska-Bartosz
Antioxidant properties of five catechins and five other flavonoids were compared with several other natural and synthetic compounds and related to glutathione and ascorbate as key endogenous antioxidants in several in vitro tests and assays involving erythrocytes. Catechins showed the highest ABTS-scavenging capacity, the highest stoichiometry of Fe(3+) reduction in the FRAP assay and belonged to the most efficient compounds in protection against SIN-1 induced oxidation of dihydrorhodamine 123, AAPH-induced fluorescein bleaching and hypochlorite-induced fluorescein bleaching...
February 15, 2018: Food Chemistry
Mariarita Stirpe, Vanessa Palermo, Michele Maria Bianchi, Romano Silvestri, Claudio Falcone, Giancarlo Tenore, Ettore Novellino, Cristina Mazzoni
BACKGROUND: During the past years, a number of studies have demonstrated the positive effect of apple on ageing and different diseases such as cancer, degenerative and cardiovascular diseases. The unicellular yeast Saccharomyces cerevisiae represents a simple eukaryotic model to study the effects of different compounds on lifespan. We previously demonstrated that apple extracts have anti-ageing effects in this organism because of their antioxidant properties. In particular, the effect is related to the presence in this fruit of polyphenols, which give a large contribution to the antioxidant activity of apples...
April 5, 2017: BMC Complementary and Alternative Medicine
Izabela Sadowska-Bartosz, Sabina Galiniak, Grzegorz Bartosz, Mariusz Zuberek, Agnieszka Grzelak, Anna Dietrich-Muszalska
The aim of this study was to compare the antioxidant activities of six atypical antipsychotic drugs: clozapine (CLZ), quetiapine, olanzapine (OLA), risperidone, ziprasidone, aripiprazole (ARI), as well as a typical antipsychotic drug, haloperidol. Several tests of antioxidant activity were used: protection of thiol groups against oxidation by peroxynitrite (PN) and 3-morpholinosydnonimine (SIN-1, generator of PN), oxidation of dihydrorhodamine 123 by PN, SIN-1 and hypochlorite (NaOCl), bleaching of fluorescein fluorescence by PN, 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH, generator of peroxyl radicals) and NaOCl, radical-scavenging activity with respect to 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) radical, 2,2-diphenyl-1-picrylhydrazyl free radical and the Ferric Reducing Antioxidant Potential...
October 2016: Schizophrenia Research
Amira Zarrouk, Thomas Nury, El-Mostafa Karym, Anne Vejux, Randa Sghaier, Catherine Gondcaille, Pierre Andreoletti, Doriane Trompier, Stéphane Savary, Mustapha Cherkaoui-Malki, Meryam Debbabi, Agnès Fromont, Jean-Marc Riedinger, Thibault Moreau, Gérard Lizard
Mitochondrial dysfunctions and oxidative stress are involved in several non demyelinating or demyelinating neurodegenerative diseases. Some of them, including multiple sclerosis (MS), are associated with lipid peroxidation processes leading to increased levels of 7-ketocholesterol (7KC). So, the eventual protective effect of dimethylfumarate (DMF), which is used for the treatment of MS, was evaluated on 7KC-treated oligodendrocytes, which are myelin synthesizing cells. To this end, murine oligodendrocytes 158N were exposed to 7KC (25, 50μM) for 24h without or with DMF (1, 25, 50μM)...
May 2017: Journal of Steroid Biochemistry and Molecular Biology
Tânia Soares, Daniela Ribeiro, Carina Proença, Renan Campos Chisté, Eduarda Fernandes, Marisa Freitas
AIMS: Silver nanoparticles (AgNPs) have emerged as an important class of nanomaterials with a wide range of industrial and medical applications. The assessment of AgNPs' biological effects in the human organism is therefore essential to evaluate the impact of these nanomaterials in public health and reassure the ratio benefit/risk. In the present study, the effect of polyvinyl pyrrolidone (PVP)-coated AgNPs with distinct sizes (10 and 50nm) on neutrophils' oxidative burst and toxicity were tested...
January 15, 2016: Life Sciences
Philip J Vernon, Leasha J Schaub, Jurandir J Dallelucca, Anthony E Pusateri, Forest R Sheppard
BACKGROUND: Maladaptive immune responses, particularly cytokine and chemokine-driven, are a significant contributor to the deleterious inflammation present in many types of injury and infection. Widely available applications to rapidly assess individual inflammatory capacity could permit identification of patients at risk for exacerbated immune responses and guide therapy. Here we evaluate neutrophil oxidative burst (NOX) capacity measured by plate reader to immuno-type Rhesus Macaques as an acute strategy to rapidly detect inflammatory capacity and predict maladaptive immune responses as assayed by cytokine array...
2015: PloS One
Yuh-Hwa Liu, Shyr-Yi Lin, Chi-Ching Lee, Wen-Chi Hou
The self-prepared pectin hydroxamic acid has been reported to have antioxidant activities [Yang, S. S., Cheng, K. D., Lin, Y. S., Liu, Y. W., & Hou, W. C. (2004). Pectin hydroxamic acids exhibit antioxidant activities in vitro. Journal of Agricultural and Food Chemistry, 52, 4270-4273]. In this study, the galacturonic acid (GalA), the monomer unit of the pectin polymer, was esterified with acidic methanol (1N HCl) at 4°C with gentle stirring for 5days to get galacturonic acid methyl ester which was further reacted with alkaline hydroxylamine to get galacturonyl hydroxamic acid (GalA-NHOH)...
July 1, 2008: Food Chemistry
Thomas Nury, Amira Zarrouk, John J Mackrill, Mohammad Samadi, Philippe Durand, Jean-Marc Riedinger, Margaux Doria, Anne Vejux, Emeric Limagne, Dominique Delmas, Michel Prost, Thibault Moreau, Mohamed Hammami, Régis Delage-Mourroux, Nora M O'Brien, Gérard Lizard
In demyelinating or non-demyelinating neurodegenerative diseases, increased levels of 7-ketocholesterol (7KC), 7β-hydroxycholesterol (7β-OHC) and 24(S)-hydroxycholesterol (24S-OHC) can be observed in brain lesions. In 158N murine oligodendrocytes, 7KC triggers a complex mode of cell death defined as oxiapoptophagy, involving simultaneous oxidative stress, apoptosis and autophagy. In these cells, 7KC as well as 7β-OHC and 24S-OHC induce a decrease of cell proliferation evaluated by phase contrast microscopy, an alteration of mitochondrial activity quantified with the MTT test, an overproduction of reactive oxygen species revealed by staining with dihydroethidium and dihydrorhodamine 123, caspase-3 activation, PARP degradation, reduced expression of Bcl-2, and condensation and/or fragmentation of the nuclei which are typical criteria of oxidative stress and apoptosis...
July 2015: Steroids
Zachary DeLoughery, Michal W Luczak, Anatoly Zhitkovich
Cr(VI) genotoxicity is caused by products of its reductive metabolism inside the cells. Reactive oxygen species (ROS) and Cr(V,IV) intermediates are potential sources of oxidative damage by Cr(VI). Here, we investigated seven fluorescent probes for the detection of ROS and non-ROS oxidants in Cr(VI) reactions with its main reducers. We found that Cr(V)-skipping metabolism of Cr(VI) by ascorbate in vitro gave no responses with all tested dyes, indicating nonreactivity of Cr(IV) and absence of ROS. Cr(VI) reduction with glutathione (GSH) or Cys strongly enhanced the fluorescence of dichlorofluorescein (DCF) and dihydrorhodamine 123 (DHR123) but produced minimal fluorescence with dihydroethidium and no increases with aminophenylfluorescein and CellRox Green, Orange, and Red...
May 19, 2014: Chemical Research in Toxicology
Markella V Zanni, Theodoros Kelesidis, Michael L Fitzgerald, Janet Lo, Suhny Abbara, Bryan Wai, Eleni Marmarelis, Nicholas J Hernandez, Otto O Yang, Judith S Currier, Steven K Grinspoon
BACKGROUND: HIV is associated with atherosclerosis and low high-density lipoprotein (HDL). With inflammation, HDL becomes dysfunctional. We previously showed that proinflammatory HDL has high HDL redox activity (HRA). In this study, we compare HRA in HIV-infected versus non-HIV-infected subjects and relate HRA to indices of macrophage activation and cardiovascular disease risk. METHODS: 102 HIV-infected subjects and 41 matched non-HIV controls without clinical cardiovascular disease underwent coronary CT angiography (CTA) and testing for immune/inflammatory biomarkers...
2014: Antiviral Therapy
E Y Ang, Y N Ang, J Y Soh, W K Liew, K W Chan, K C Thoon, C Y Chong, Y L Lau, B W Lee
BACKGROUND: Dihydrorhodamine (DHR) flow cytometric analysis is used to evaluate granulocyte oxidative bursts and is the test of choice for the diagnosis of chronic granulomatous disease (CGD). We present the clinical and DHR test profiles of five subjects assessed during and after acute illness. METHODS: This was a retrospective report of the findings of five out of a total of one hundred and seventeen patients, whose blood was sent to the laboratory for dihydrorhodamine-123 flow cytometry testing between January 2005 and December 2010...
2013: Clinical Laboratory
Victor J Johnson, Masashi Tsunoda, Thomas F Murray, Raghubir P Sharma
Effects of aluminum (Al) on membrane properties of excitable cells are not fully understood. Several reports have identified cellular membranes as sensitive targets for Al intoxication. In the present study, we tested the hypothesis that treatment with Al would alter membrane fluidity and potential and these changes would correlate with aberrant generation of cellular oxidants. The effects of in vitro Al exposure in resting rat pheochromocytoma (PC-12) cells, a model that exhibits neuron-like properties, were investigated...
February 2005: Environmental Toxicology and Pharmacology
Jiřina Procházková, Lukáš Kubala, Hana Kotasová, Iva Gudernová, Zuzana Šrámková, Michaela Pekarová, Balázs Sarkadi, Jiří Pacherník
Intracellular production of reactive oxygen species (ROS) plays an important role in the control of cell physiology. For the assessment of intracellular ROS production, a plethora of fluorescent probes is commonly used. Interestingly, chemical structures of these probes imply they could be substrates of plasma membrane efflux pumps, called ABC transporters. This study tested whether the determination of intracellular ROS production and mitochondrial membrane potential by selected fluorescent probes is modulated by the expression and activity of ABC transporters...
July 2011: Free Radical Research
Ludwig K Groebler, Joe Liu, Anu Shanu, Rachel Codd, Paul K Witting
Accumulating Mb (myoglobin) in the kidney following severe burns promotes oxidative damage and inflammation, which leads to acute renal failure. The potential for haem-iron to induce oxidative damage has prompted testing of iron chelators [e.g. DFOB (desferrioxamine B)] as renal protective agents. We compared the ability of DFOB and a DFOB-derivative {DFOB-AdAOH [DFOB-N-(3-hydroxyadamant-1-yl)carboxamide]} to protect renal epithelial cells from Mb insult. Loading kidney-tubule epithelial cells with dihydrorhodamine-123 before exposure to 100 μM Mb increased rhodamine-123 fluorescence relative to controls (absence of Mb), indicating increased oxidative stress...
May 1, 2011: Biochemical Journal
Hugh Farrell, Jennifer Hayes, John Laffey, Neil Rowan
This constitutes the first study to report on the relationship between pulsed UV light (PL) irradiation and the simultaneous occurrence of molecular and cellular damage in clinical strains of Candida albicans. Microbial protein leakage and propidium iodide (PI) uptake assays demonstrated significant increases in cell membrane permeability in PL-treated yeast that depended on the amount of UV pulses applied. This finding correlated well with the measurement of increased levels of lipid hydroperoxidation in the cell membrane of PL-treated yeast...
February 2011: Journal of Microbiological Methods
Binu Tharakan, Felicia A Hunter, W Roy Smythe, Ed W Childs
1. Oxidative stress induced by reactive oxygen species (ROS) is a key mediator of haemorrhagic shock (HS)-induced vascular hyperpermeability. In the present study, curcumin, a natural anti-oxidant obtained from turmeric (Curcuma longa), was tested against HS-induced hyperpermeability and associated ROS formation in rat mesenteric post-capillary venules in vivo and in rat lung microvascular endothelial cells (RLMEC) in vitro. 2. In rats, HS was induced by withdrawing blood to reduce mean arterial pressure to 40 mmHg for 60 min, followed by resuscitation for 60 min...
September 2010: Clinical and Experimental Pharmacology & Physiology
M C Wiggins, A R Woolums, D J Hurley, S Sanchez, D T Ensley, D Donovan
Mycoplasma bovis (M. bovis) contributes to a number of clinical syndromes in cattle; in particular, chronic pneumonia that is poorly responsive to therapy has been increasingly recognized as an important cause of morbidity, mortality, and financial loss. M. bovis impairs host immune function, but little is known about whether field isolates vary significantly in their effect on immune function. This research tested the hypothesis that different field isolates vary in their ability to suppress cellular metabolism and cellular production of radical oxygen species (ROS) by bovine leukocytes...
January 2011: Comparative Immunology, Microbiology and Infectious Diseases
Lavinia Bhatt, Gillian Groeger, Kieran McDermott, Thomas G Cotter
PURPOSE: The production of reactive oxygen species (ROS) can lead to oxidative stress, which is a strong contributory factor to many ocular diseases. In this study, the removal of trophic factors is used as a model system to investigate the effects of stress in the retina. The aims were to determine if both rod and cone photoreceptor cells produce ROS when they are deprived of trophic factor support and to demonstrate if the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (Nox) enzymes are responsible for this ROS production...
2010: Molecular Vision
Elina Pathak, Philip R Mayeux
Sepsis-induced acute kidney injury (AKI) is a complex disease characterized by generation of inducible nitric oxide synthase (iNOS)-derived reactive nitrogen species (RNS) by the renal tubular epithelium. While most in vitro models of sepsis use combinations of lipopolysaccharide and cytokines to simulate exposure to inflammatory mediators thought to play a role in sepsis, the relevance of these models is limited. To address the need for a model that more closely mimics the tubular microenvironment during sepsis, we developed an in vitro model where mIMCD-3 (murine tubular epithelial) cells are treated with media containing 5% serum collected from mice at 4 h after cecal ligation and puncture (CLP) or sham surgery (no sepsis)...
June 2010: Toxicological Sciences: An Official Journal of the Society of Toxicology
Marisa Freitas, Graça Porto, José L F C Lima, Eduarda Fernandes
The evaluation of reactive oxygen species (ROS) and reactive nitrogen species (RNS) production by neutrophils is currently a matter of extensive research, with scientific reports showing an enormous variability on the detection methods as well as the concentration of the detecting probes. Also the incubation media used to test neutrophils and the respective ionic concentration, as well as the glucose concentration, varies enormously from study to study. This variability often results in different sensibility and/or response of neutrophils to stimulating agents, which can be a focus of confounding and sometime contradictory results among reports...
June 15, 2009: Talanta
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