Superoxide () is a reactive oxygen species (ROS) that is primarily produced by the one‐electron transfer of photooxidized chromophoric dissolved organic matter (CDOM) to O2in sunlit natural waters. Here we examine the environmental and chemical parameters (pH, ionic strength, buffer, and halides) that may influence photochemical production rates and decay pathways in natural water. Using the enzyme superoxide dismutase and H2O2measurements, we present results from an irradiated freshwater CDOM source indicating that reductive decay pathways (
In this study, we investigated the effects of weak static magnetic fields (SMFs) on HT‐1080 human fibrosarcoma cells. Exposures to SMFs for four consecutive days were varied from 0.5 to 600 µT for treated units, while exposures to control units were held at 45 µT. Growth rates were measured by comparing cell counts, whereas membrane potentials, mitochondrial calcium, mitochondrial superoxide (O2−), nitric oxide (NO), hydrogen peroxide (H2O2), intercellular pH, and oxidative stress were measured by using fluorescent dyes. The relative cell growth rates vary with the angle of the SMFs. Increases in the magnitude of the SMFs increased concentrations of mitochondrial calcium and membrane potential and decreased intracellular pH. H2O2, an important reactive oxygen species (ROS), increases at 100 and 200 µT, decreases at 300 and 400 µT and increases again at 500 and 600 µT. Overall, oxidative stress increases slightly with increasing SMFs, while superoxide and NO concentrations decrease. These results indicate that weak SMFs can accelerate and inhibit cell growth rates and induce alterations in ROS. Changes in ROS and oxidative stress are important for various cell functions. Calcium influx into mitochondria was one of the initial steps into the corresponding changes. Bioelectromagnetics. 2021. © 2021 Bioelectromagnetics Society
more » « less- NSF-PAR ID:
- 10256233
- Publisher / Repository:
- Wiley Blackwell (John Wiley & Sons)
- Date Published:
- Journal Name:
- Bioelectromagnetics
- Volume:
- 42
- Issue:
- 3
- ISSN:
- 0197-8462
- Page Range / eLocation ID:
- p. 212-223
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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Abstract P /P SOD) dominate with increased pH and NaCl additions and maximal photoproduction rates () increase with carbonate compared to borate buffer. Over 2 h of irradiation, a significant decline in was seen for all samples along with a minor increase in oxidative pathways. These results imply shifts in decay pathways and production rates that seem to vary across natural waters and as a function of irradiation history. -
In response to various pathological stimuli, such as oxidative and energy stress accompanied by high Ca 2+ , mitochondria undergo permeability transition (PT) leading to the opening of the non-selective PT pores (PTP) in the inner mitochondrial membrane. Opening of the pores at high conductance allows the passage of ions and solutes <1.5 kD across the membrane, that increases colloid osmotic pressure in the matrix leading to excessive mitochondrial swelling. Calcium retention capacity (CRC) reflects maximum Ca 2+ overload of mitochondria that occurs just before PTP opening. Quantification of CRC is important for elucidating the effects of different pathological stimuli and the efficacy of pharmacological agents on the mitochondria. Here, we performed a comparative analysis of CRC in mitochondria isolated from H9c2 cardioblasts, and in permeabilized H9c2 cells in situ to highlight the strengths and weaknesses of the CRC technique in isolated cell mitochondria vs. permeabilized cells. The cells were permeabilized by digitonin or saponin, and the Ca 2+ -sensitive fluorescence probe Calcium Green-5N was used in both preparations. Results demonstrated the interference of dye-associated fluorescence signals with saponin and the adverse effects of digitonin on mitochondria at high concentrations. Analysis of the CRC in permeabilized cells revealed a higher CRC in the saponin-permeabilized cells in comparison with the digitonin-permeabilized cells. In addition, the mitochondrial CRC in saponin-permeabilized cells was higher than in isolated mitochondria. Altogether, these data demonstrate that the quantification of the mitochondrial CRC in cultured cells permeabilized by saponin has more advantages compared to the isolated mitochondria.more » « less
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Abstract Background Response to oxidative stress is universal in almost all organisms and the mitochondrial membrane protein, BbOhmm, negatively affects oxidative stress responses and virulence in the insect fungal pathogen,
Beauveria bassiana . Nothing further, however, is known concerning howBbOhmm and this phenomenon is regulated.Results Three
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Vascular cells develop resilience to H2O2during ageing by preventing Ca2+overload and endothelial integrity promotes SMC survival.
Abstract Advanced age is associated with elevated oxidative stress and can protect the endothelium from cell death induced by H2O2. Whether such protection occurs for intact vessels or differs between smooth muscle cell (SMC) and endothelial cell (EC) layers is unknown. We tested the hypothesis that ageing protects SMCs and ECs during acute exposure to H2O2(200 µ
m , 50 min). Mouse superior epigastric arteries (SEAs; diameter, ∼150 µm) were isolated and pressurized to 100 cmH2O at 37˚C. For SEAs from young (4 months) mice, H2O2killed 57% of SMCs and 11% of ECs in malesvs . 8% and 2%, respectively, in females. Therefore, SEAs from males were studied to resolve the effect of ageing and experimental interventions. For old (24 months) mice, SMC death was reduced to 10% with diminished accumulation of [Ca2+]iin the vessel wall during H2O2exposure. In young mice, genetic deletion of IL‐10 mimicked the protective effect of ageing on cell death and [Ca2+]iaccumulation. Whereas endothelial denudation or gap junction inhibition (carbenoxolone; 100 µm ) increased SMC death, inhibiting NO synthase (l ‐NAME, 100 µm ) or scavenging peroxynitrite (FeTPPS, 5 µm ) reduced SMC death along with [Ca2+]i. Despite NO toxicity via peroxynitrite formation, endothelial integrity protects SMCs. Caspase inhibition (Z‐VAD‐FMK, 50 µm ) attenuated cell death with immunostaining for annexin V, cytochrome C, and caspases 3 and 9 pointing to induction of intrinsic apoptosis during H2O2exposure. We conclude that advanced age reduces Ca2+influx that triggers apoptosis, thereby promoting resilience of the vascular wall during oxidative stress. -
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Abstract Trace metals play a critical role in the development of culture media used for the production of therapeutic proteins. Iron has been shown to enhance the productivity of monoclonal antibodies during Chinese hamster ovary (CHO) cell culture. However, the redox activity and pro‐oxidant behavior of iron may also contribute toward the production of reactive oxygen species (ROS). In this work, we aim to clarify the influence of trace iron by examining the relationship between iron supplementation to culture media, mAb productivity and glycosylation, and oxidative stress interplay within the cell. Specifically, we assessed the impacts of iron supplementation on (a) mAb production and glycosylation; (b) mitochondria‐generated free hydroxyl radicals (ROS); (c) the cells ability to store energy during oxidative phosphorylation; and (d) mitochondrial iron concentration. Upon the increase of iron at inoculation, CHO cells maintained a capacity to rebound from iron‐induced viability lapses during exponential growth phase and improved mAb productivity and increased mAb galactosylation. Fluorescent labeling of the mitochondrial hydroxyl radical showed enhanced environments of oxidative stress upon iron supplementation. Additional labeling of active mitochondria indicated that, despite the enhanced production of ROS in the mitochondria, mitochondrial membrane potential was minimally impacted. By replicating iron treatments during seed train passaging, the CHO cells were observed to adapt to the shock of iron supplementation prior to inoculation. Results from these experiments demonstrate that CHO cells have the capacity to adapt to enhanced environments of oxidative stress and improve mAb productivity and mAb galactosylation with minimal perturbations to cell culture.
