Microtiter plate assay for the measurement of glutathione and glutathione disulfide in large numbers of biological samples
Abstract
By combining the least complicated and expedient methods of sample handling with the sensitivity and specificity of the GSH assay by enzymatic recycling and the small volumes and software capabilities of microtiter plate technology we have devised a rapid, sensitive, and easy assay for GSH and GSSG in biological samples. The assay is sensitive to 5 pmol in sample volumes of 50 μl, although other volumes could be used. The use of a computer-driven microplate with software capable of linear kinetic data storage and analysis on each well, Maxline series microplate readers and Softmax software, enables the user not only to assay large numbers of samples per day but also to have immediate calculated results. We suggest by examples that measurements of total GSH as well as changes in GSH:GSSG in vitro and in vivo are feasible with this technology.
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Variations of oxidative potential of PM<inf>2.5</inf> in a medium-sized residential city in South Korea measured using three different chemical assays
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Silver nanoparticles from Ocimum basilicum L. tea: A green route with potent anticancer efficacy
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Neuroprotective effects of Petiveria alliacea on scopolamine-induced learning and memory impairment mouse model
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To assess the effects of the P. alliacea leaves methanol fraction (PMF) on a scopolamine-induced learning and memory impairment mouse model related to acetylcholinesterase activity and oxidative stress.
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PMF led to memory improvement in the behavioral tests in mice with scopolamine-induced cognitive impairment. Moreover, PMF inhibited acetylcholinesterase activity and showed antioxidant potential that in turn attenuated cholinergic degradation. Additionally, PMF increased glutathione levels and glutathione reductase activity and reduced MDA levels in the brain. Moreover, no acute toxicity was detected with the use of PMF.
In a mouse model of scopolamine-induced cognitive deficit, PMF exhibited protective effects, decreasing oxidative damage and regulating cholinergic function in the brain bearing significant memory enhancing potency. These data suggest that PMF is a promising candidate for developing therapies for neurodegenerative disorders.
Induced swimming modified the antioxidant status of gilthead seabream (Sparus aurata)
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Dietary tributyrin supplementation enhances the immune and antioxidant responses of rainbow trout (Oncorhynchus mykiss) without changes in fish performance
2023, Aquaculture ReportsDietary supplementation with tributyrin (TBT), a source of butyric acid, has been shown to improve growth performance and health status when high levels of plant ingredients are included in aquafeeds. Here, we investigated the response of rainbow trout (Oncorhynchus mykiss) to a plant-based diet supplemented with TBT aiming to improve fish performance and welfare. Juvenile rainbow trout (19.0 ± 0.19 g, mean ± SEM) were divided into 12 tanks (210 L, 40 fish/tank) connected to a RAS system and fed four experimental diets in triplicate to apparent satiety. Diets tested comprised a basal diet (CTR) including 10% fishmeal and 10% fish oil, plus 83% plant-derived ingredients (44% crude protein, 18% crude fat, gross energy 21.8 MJ kg−1) with increasing inclusion levels of TBT (55% purity), supplying 0.05% (TBT1), 0.1% (TBT2) and 0.2% (TBT4) of this compound. Fish performance was evaluated after 44 days. Following the growth trial, the effect of the diets on selected plasma innate immune and hepatic antioxidant parameters of trout was investigated at 3 h and 24 h after feeding. TBT supplementation had no significant impact on growth performance, feed efficiency, feed intake, or proximal composition in rainbow trout (P > 0.05). However, the plasma lysozyme activity was higher in fish fed the TBT2 diet than in those fed CTR or TBT4 diets after 3 h (P = 0.021). In addition, the plasma antiprotease activity in fish fed the TBT2 and TBT4 diets was higher than those fed the CTR diet after 24 h (P = 0.010 and 0.013, respectively). Catalase activity in the liver of trout fed the TBT4 diet was higher than in fish fed CTR or TBT2 diets after 3 h (P = 0.016 and 0.019). Results showed that effective levels of dietary TBT supplementation at 0.1% and 0.2% could result in improved immune and antioxidant responses in trout, respectively. Therefore, dietary TBT supplementation in this carnivorous species may result in enhanced welfare when high levels of plant-derived ingredients are used in aquafeeds.
Comparison of toxicity of silver nanomaterials and silver nitrate on developing zebrafish embryos: Bioavailability, osmoregulatory and oxidative stress
2023, ChemosphereThe mechanisms of toxicity of engineered nanomaterials (ENMs) to the early life stages of freshwater fish, and the relative hazard compared to dissolved metals, is only partially understood. In the present study, zebrafish (Danio rerio) embryos were exposed to lethal concentrations of silver nitrate (AgNO3) or silver (Ag) ENMs (primary size 42.5 ± 10.2 nm). The 96 h-LC50 for AgNO3 was 32.8 ± 0.72 μg Ag L−1 (mean ± 95% CI) compared to 6.5 ± 0.4 mg L−1 of the whole material for Ag ENMs; with the ENMs being orders of magnitude less toxic than the metal salt. The EC50 for hatching success was 30.5 ± 1.4 μg Ag L−1 and 6.04 ± 0.4 mg L−1 for AgNO3 and Ag ENMs, respectively. Further sub-lethal exposures were performed with the estimated LC10 concentrations for both AgNO3 or Ag ENMs over 96 h, where about 3.7% of the total Ag as AgNO3 was internalised, as measured by Ag accumulation in the dechorionated embryos. However, for the ENM exposures, nearly all (99.8%) of the total Ag was associated with chorion; indicating the chorion as an effective barrier to protect the embryo in the short term. Calcium (Ca2+) and sodium (Na+) depletion was induced in embryos by both forms of Ag, but hyponatremia was more pronounced in the nano form. Total glutathione (tGSH) levels declined in embryos exposed to both Ag forms, but a superior depletion occurred with the nano form. Nevertheless, oxidative stress was mild as superoxide dismutase (SOD) activity stayed uniform and the sodium pump (Na+/K+-ATPase) activity had no appreciable inhibition compared to the control. In conclusion, AgNO3 was more toxic to the early life stage zebrafish than the Ag ENMs, still differences were found in the exposure and toxic mechanisms of both Ag forms.