Monochlorobimane Fluorometric Method to Measure Tissue Glutathione

Kamencic, Huse; Lyon, Andrew W.; Paterson, Phyllis G.; Juurlink, Bernhard H.J.

doi:10.1006/abio.2000.4765

Cited by 233 publications

(156 citation statements)

References 15 publications

(7 reference statements)

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“…One method used to determine GSH levels in living cells is based on the GSH-sensitive probe mCB. Glutathione S-transferases form GSH-mCB complexes that can be measured fluorometrically [45]. Treatment with various concentrations of DMSA-MNP led to a non-significant decrease in GSH levels, whereas 750 µM H 2 O 2 treatment reduced GSH to 28% of control levels (Fig.…”

Section: In Vitro Dmsa-mnp Cytotoxicitymentioning

confidence: 98%

Long term biotransformation and toxicity of dimercaptosuccinic acid-coated magnetic nanoparticles support their use in biomedical applications

Mejías

Gutiérrez

Salas

et al. 2013

Journal of Controlled Release

114

101

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Although iron oxide magnetic nanoparticles (MNP) have been proposed for numerous biomedical applications, little is known about their biotransformation and long-term toxicity in the body. Dimercaptosuccinic acid (DMSA)-coated magnetic nanoparticles have been proven efficient for in vivo drug delivery, but these results must nonetheless be sustained by comprehensive studies of long-term distribution, degradation and toxicity. We studied DMSA-coated magnetic nanoparticles effects in vitro on NCTC 1469 nonparenchymal hepatocytes, and analyzed their biodistribution and biotransformation in vivo in C57BL/6 mice. Our results indicate that DMSA-coated magnetic nanoparticles have little effect on cell viability, oxidative stress, cell cycle or apoptosis on NCTC 1469 cells in vitro. In vivo distribution and transformation was studied by alternating current magnetic susceptibility measurements, a technique that permits distinction of MNP from other iron species. Our results show that DMSA-coated MNP accumulate in spleen, liver and lung tissues for extended periods of time, in which nanoparticles undergo a process of conversion from superparamagnetic iron oxide nanoparticles to other nonsuperparamagnetic iron forms, with no significant signs of toxicity.

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Section: In Vitro Dmsa-mnp Cytotoxicitymentioning

confidence: 98%

Long term biotransformation and toxicity of dimercaptosuccinic acid-coated magnetic nanoparticles support their use in biomedical applications

Mejías

Gutiérrez

Salas

et al. 2013

Journal of Controlled Release

114

101

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“…Intracellular GSH levels were assayed as described [33]. Briefly, 100 μl of ice-cold lysis buffer was added to T cells (1×10 6 ), incubated on ice for 10 min, and centrifuged at 12,000×g for 10 min in a microfuge.…”

Section: Measurement Of Intracellular Gshmentioning

confidence: 99%

Redox regulation of the proteasome in T lymphocytes during aging

Das

Ponnappan

2007

Free Radical Biology and Medicine

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Proteasome is a major cellular organelle responsible for the regulated turn-over of both normal and misfolded proteins. Recent reports from our laboratory have implicated lowered proteasomal chymotryptic activity to be responsible for decreased induction of the transcription factor NFκB in T lymphocytes during aging. In this study, we have further analyzed the basis for this decline in proteasomal function, by focusing on the role of oxidative stress. Upon exposure to the pro-oxidant BSO, both ATP-stimulatable 26S, as well as ATP-independent 20S proteasomal catalytic activity could be down-regulated in T cells from young donors, mimicking the decline observed in T cells from the elderly. Loss in these catalytic activities, following exposure to pro-oxidant stimulus also resulted in a decline in both activation-induced proliferation as well as degradation of the inhibitor IκBα, with concomitant increase in the accumulation of carbonylated proteins, mimicking responses seen in T cells from the elderly. Pretreatment with an antioxidant, NAC could override pro-oxidantmediated, but not age-associated decrease in both 20S proteasomal activities. These results suggest that the decrease in proteasomal activities observed during aging may be secondary to oxidative stress and underlie immune senescence.

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“…Thereafter, the fluorescence of each supernatant was measured (l ex ¼ 380 nm, l em ¼ 470 nm) using a fluorescence reader (Spectra MAX Gemini; Molecular Devices). As a standard we used GSH dissolved in MTP medium in a range 0-100 mM (Kamencic et al, 2000). All data are expressed as mean ± SEM.…”

Section: Determination Of Reduced Gsh Contentmentioning

confidence: 99%

N-acetyl Cysteine Treatment Rescues Cognitive Deficits Induced by Mitochondrial Dysfunction in G72/G30 Transgenic Mice

Otte

Sommersberg

Kudin

et al. 2011

Neuropsychopharmacol

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Genetic studies have implicated the evolutionary novel, anthropoid primate-specific gene locus G72/G30 in psychiatric diseases. This gene encodes the protein LG72 that has been discussed to function as a putative activator of the peroxisomal enzyme D-aminoacid-oxidase (DAO) and as a mitochondrial protein. We recently generated 'humanized' bacterial artificial chromosome transgenic mice (G72Tg) expressing G72 transcripts in cells throughout the brain. These mice exhibit several behavioral phenotypes related to psychiatric diseases. Here we show that G72Tg mice have a reduced activity of mitochondrial complex I, with a concomitantly increased production of reactive oxygen species. Affected neurons display deficits in short-term plasticity and an impaired capability to sustain synaptic activity. These deficits lead to an impairment in spatial memory, which can be rescued by pharmacological treatment with the glutathione precursor N-acetyl cysteine. Our results implicate LG72-induced mitochondrial and synaptic defects as a possible pathomechanism of psychiatric disorders.

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Monochlorobimane Fluorometric Method to Measure Tissue Glutathione

Cited by 233 publications

References 15 publications

Long term biotransformation and toxicity of dimercaptosuccinic acid-coated magnetic nanoparticles support their use in biomedical applications

Long term biotransformation and toxicity of dimercaptosuccinic acid-coated magnetic nanoparticles support their use in biomedical applications

Redox regulation of the proteasome in T lymphocytes during aging

N-acetyl Cysteine Treatment Rescues Cognitive Deficits Induced by Mitochondrial Dysfunction in G72/G30 Transgenic Mice

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