Use of spectroscopic probes for detection of reactive oxygen species

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Abstract:The erythrocytes of 12-month old Sod1 -/-mice showed an increased level of reactive oxygen species (ROS), as estimated by the degree of dihydroethidine and dihydrorhodamine oxidation, and the increased level of Heinz bodies. No indices of severe oxidative stress were found in the red blood cells and blood plasma of Sod1 -/-mice as judged from the lack of significant changes in the levels of erythrocyte and plasma glutathione, plasma protein thiol and carbonyl groups and thiobarbituric-acid reactive substances in the blood plasma. However, a decreased erythrocyte lifespan, increased reticulocyte count and splenomegaly were noted, indicating the importance of superoxide dismutase for maintaining erythrocyte viability. The levels of erythrocyte ROS and Sod1 +/-mice, suggesting that a superoxide dismutase activity decrease to half of its normal value may be sufficient to secure the protective effects of the enzyme.

Section: Discussionsupporting

confidence: 64%

The effects of superoxide dismutase knockout on the oxidative stress parameters and survival of mouse erythrocyt

Grzelak¹,

Kruszewski²,

Macierzyńska³

et al. 2009

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“…After a 10-min incubation at the ambient temperature, the fluorescence intensity of the oxidation product (rhodamine 123) was measured. H 2 R 123 is known to be oxidized to the fluorescent compound rhodamine 123 by hydrogen peroxide and organic peroxides in reactions catalyzed by peroxidases or ferric ions, but also by a range of other oxidants [20,21]. Though the identification of specific oxidants of H 2 R 123 in a cell is not easy, the rate of oxidation is usually assumed to reflect the overall level of the reactive oxygen species.…”

Section: Resultsmentioning

confidence: 99%

“…Though the identification of specific oxidants of H 2 R 123 in a cell is not easy, the rate of oxidation is usually assumed to reflect the overall level of the reactive oxygen species. However, such an interpretation may be too straightforward, since the measured rate of dihydrorhodamine 123 oxidation is a resultant of the rate of ROS production and the level of endogenous antioxidants competing with the probe for the ROS [22,23]. The growth medium (i.e.…”

Section: Resultsmentioning

confidence: 99%

The effect of growth medium on the antioxidant defense of Saccharomyces cerevisiae

Macierzyńska

Grzelak

Bartosz

2007

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Abbreviations used: DTNB -5,5'-dithiobis-(2-nitrobenzoic acid); H 2 R 123 -dihydrorhodamine 123; MES -2-morpholinoethanesulfonic acid; ROS -reactive oxygen species; SOD -superoxide dismutase; YPD medium -1% yeast extract, 1% Bacto-peptone, 2% glucose, YPG medium, 1% yeast extract, 1% Bacto-peptone,2% glycerol; YPE medium -1% yeast extract, 1% Bacto-peptone, 3% ethanol Abstract: We compared the oxidation of dihydrorhodamine 123, glutathione contents and activities of superoxide dismutase (SOD) and catalase for three wild-type strains of Saccharomyces cerevisiae grown on media with different carbon sources. The rate of oxidation of dihydrorhodamine 123 was much higher in respiring cells grown on ethanol or glycerol media than in fermenting cells grown on glucose medium. The total SOD activity was highest on glycerol medium and lowest on ethanol medium, while the catalase activity was highest on glycerol medium. The sequence of glutathione content values was: glucose > ethanol > glycerol.

“…H 2 DCFDA does not fluoresce until it enters the cell, where the acetate groups are removed by intracellular esterases to form H 2 DCF. H 2 DCF is then oxidized to fluorescent DCF [34]. Cells were seeded in 12-well plates at a density of 25 × 10 4 cells/well in DMEM medium and incubated for 20 h. After 24 h dendrimer treatment, the supernatant was discarded, and the cells were washed with PBS and stained with 2.5 μM H 2 DCFDA for 15 min in growing conditions.…”

Section: Measurement Of Reactive Oxygen Speciesmentioning

confidence: 99%

Viologen-phosphorus dendrimers exhibit minor toxicity against a murine neuroblastoma cell line

Łaźniewska

Miłowska

Katir

et al. 2013

Dendrimers containing viologen (derivatives of 4,4'-bipyridyl) units in their structure have been demonstrated to exhibit antiviral activity against human immunodeficiency virus (HIV-1). It has also recently been revealed that novel dendrimers with both viologen units and phosphorus groups in their structure show different antimicrobial, cytotoxic and hemotoxic properties, and have the ability to influence the activity of cholinesterases and to inhibit α-synuclein fibrillation. Since the influence of viologen-phosphorus structures on basic cellular processes had not been investigated, we examined the impact of such macromolecules on the murine neuroblastoma cell line (N2a). We selected three water-soluble viologen-phosphorus (VPD) dendrimers, which differ in their core structure, number of viologen units and number and type of surface groups, and analyzed several aspects of the cellular response. These included cell viability, generation of reactive oxygen species (ROS), alterations in mitochondrial activity, morphological modifications, and the induction of apoptosis and necrosis. The MTT assay results suggest that all of the tested dendrimers are only slightly cytotoxic. Although some changes in ROS formation and mitochondrial function were detected, the three compounds did not induce apoptosis or necrosis. In light of these results, we can assume that the tested VPD are relatively safe for mouse neuroblastoma cells. Although more research on their safety is needed, VPD seem to be promising nanoparticles for further biomedical investigation.