Morpho‐metabotyping the oxidative stress response

AbstractOxidative stress and reactive oxygen species (ROS) are central to many physiological and pathophysiological processes. However, due to multiple technical challenges, it is hard to capture a comprehensive readout of the cell, involving both biochemical and functional status. We addressed this problem by developing a fully parallelized workflow for metabolomics (providing absolute quantities for > 100 metabolites including TCA cycle, pentose phosphate pathway, purine metabolism, glutathione metabolism, cysteine and methionine metabolism, glycolysis and gluconeogenesis) and live cell imaging microscopy. The correlative imaging strategy was applied to study morphological and metabolic adaptation of cancer cells upon short-term hydrogen peroxide (H2O2) exposure in vitro. The combination provided rich metabolic information at the endpoint of exposure together with imaging of mitochondrial effects. As a response, superoxide concentrations were elevated with a strong mitochondrial localization, and multi-parametric image analysis revealed a shift towards fragmentation. In line with this, metabolism reflected both the impaired mitochondrial function and shifts to support the first-line cellular defense and compensate for energy loss. The presented workflow combining high-end technologies demonstrates the applicability for the study of short-term oxidative stress, but it can be suitable for the in-depth study of various short-term oxidative and other cellular stress-related phenomena.

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Resolving the TCA cycle and pentose-phosphate pathway of Clostridium acetobutylicum ATCC 824: Isotopomer analysis, in vitro activities and expression analysis

Biotechnology Journal ◽

10.1002/biot.201000282 ◽

2010 ◽

Vol 6 (3) ◽

pp. 300-305 ◽

Cited By ~ 69

Author(s):

Scott B. Crown ◽

Dinesh C. Indurthi ◽

Woo Suk Ahn ◽

Jungik Choi ◽

Eleftherios T. Papoutsakis ◽

...

Keyword(s):

Expression Analysis ◽

Pentose Phosphate Pathway ◽

Clostridium Acetobutylicum ◽

Tca Cycle ◽

Pentose Phosphate ◽

The Tca Cycle ◽

Isotopomer Analysis ◽

Clostridium Acetobutylicum Atcc 824

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Oxidative Stress, Glutathione Metabolism, and Liver Regeneration Pathways Are Activated in Hereditary Tyrosinemia Type 1 Mice upon Short-Term Nitisinone Discontinuation

Genes ◽

10.3390/genes12010003 ◽

2020 ◽

Vol 12 (1) ◽

pp. 3

Author(s):

Haaike Colemonts-Vroninks ◽

Jessie Neuckermans ◽

Lionel Marcelis ◽

Paul Claes ◽

Steven Branson ◽

...

Keyword(s):

Oxidative Stress ◽

Stress Response ◽

Liver Regeneration ◽

The Body ◽

Glutathione Metabolism ◽

Short Term ◽

Therapeutic Regime ◽

Tyrosinemia Type 1 ◽

Hereditary Tyrosinemia

Hereditary tyrosinemia type 1 (HT1) is an inherited condition in which the body is unable to break down the amino acid tyrosine due to mutations in the fumarylacetoacetate hydrolase (FAH) gene, coding for the final enzyme of the tyrosine degradation pathway. As a consequence, HT1 patients accumulate toxic tyrosine derivatives causing severe liver damage. Since its introduction, the drug nitisinone (NTBC) has offered a life-saving treatment that inhibits the upstream enzyme 4-hydroxyphenylpyruvate dioxygenase (HPD), thereby preventing production of downstream toxic metabolites. However, HT1 patients under NTBC therapy remain unable to degrade tyrosine. To control the disease and side-effects of the drug, HT1 patients need to take NTBC as an adjunct to a lifelong tyrosine and phenylalanine restricted diet. As a consequence of this strict therapeutic regime, drug compliance issues can arise with significant influence on patient health. In this study, we investigated the molecular impact of short-term NTBC therapy discontinuation on liver tissue of Fah-deficient mice. We found that after seven days of NTBC withdrawal, molecular pathways related to oxidative stress, glutathione metabolism, and liver regeneration were mostly affected. More specifically, NRF2-mediated oxidative stress response and several toxicological gene classes related to reactive oxygen species metabolism were significantly modulated. We observed that the expression of several key glutathione metabolism related genes including Slc7a11 and Ggt1 was highly increased after short-term NTBC therapy deprivation. This stress response was associated with the transcriptional activation of several markers of liver progenitor cells including Atf3, Cyr61, Ddr1, Epcam, Elovl7, and Glis3, indicating a concreted activation of liver regeneration early after NTBC withdrawal.

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Can C-phycocyanin be used as a sperm protector?

Veterinaria México OA ◽

10.22201/fmvz.24486760e.2018.4.642 ◽

2019 ◽

Vol 5 (4) ◽

Author(s):

Emma Gabriela Antonio Marcos ◽

O. Monroy Hermosillo ◽

E. Cortés Barberena ◽

E. Rodríguez Tobón ◽

J. G. Rivera Martínez ◽

...

Keyword(s):

Oxidative Stress ◽

Sus Scrofa ◽

Antioxidant Properties ◽

Medical Applications ◽

Protein Supplement ◽

Oxygen Species ◽

Short Term ◽

Progressive Motility ◽

Sus Scrofa Domesticus

C-phycocyanin (C-PC) is a biocompound extracted from Arthrospira maxima. It is a chromophore-protein complex, with antioxidant properties. Its ability to prevent oxidative stress allows for diverse medical applications. This study evaluates the use of C-PC as a protein supplement and an antioxidant for in-vitro sperm preservation in a short-term extender. Viability, progressive motility, DNA damage and percentage of reactive oxygen species where assessed in Swine (Sus scrofa domesticus) sperm stored for up to 72 hours at 4 °C. Treatments with C-PC had the following concentrations: 0, 34.5, 69, 138 and 207 μg mL−1. Progressive motility and percentage of sperm with undamaged DNA were unchanged (20%) after storage for 48 hours using the 138 μg mL C-PC concentration−1.

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Beneficial effects of acute inhibition of the oxidative pentose phosphate pathway in the failing heart

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00783.2013 ◽

2014 ◽

Vol 306 (5) ◽

pp. H709-H717 ◽

Cited By ~ 13

Author(s):

Claudio Vimercati ◽

Khaled Qanud ◽

Gianfranco Mitacchione ◽

Danuta Sosnowska ◽

Zoltan Ungvari ◽

...

Keyword(s):

Oxidative Stress ◽

Pentose Phosphate Pathway ◽

Tissue Concentration ◽

Glucose Consumption ◽

Pentose Phosphate ◽

Oxidative Pentose Phosphate Pathway ◽

Stroke Work ◽

Failing Heart

In vitro studies suggested that glucose metabolism through the oxidative pentose phosphate pathway (oxPPP) can paradoxically feed superoxide-generating enzymes in failing hearts. We therefore tested the hypothesis that acute inhibition of the oxPPP reduces oxidative stress and enhances function and metabolism of the failing heart, in vivo. In 10 chronically instrumented dogs, congestive heart failure (HF) was induced by high-frequency cardiac pacing. Myocardial glucose consumption was enhanced by raising arterial glycemia to levels mimicking postprandial peaks, before and after intravenous administration of the oxPPP inhibitor 6-aminonicotinamide (80 mg/kg). Myocardial energy substrate metabolism was measured with radiolabeled glucose and oleic acid, and cardiac 8-isoprostane output was used as an index of oxidative stress. A group of five chronically instrumented, normal dogs served as control. In HF, raising glycemic levels from ∼80 to ∼170 mg/dL increased cardiac isoprostane output by approximately twofold, whereas oxPPP inhibition normalized oxidative stress and enhanced cardiac oxygen consumption, glucose oxidation, and stroke work. In normal hearts glucose infusion did not induce significant changes in cardiac oxidative stress. Myocardial tissue concentration of 6P-gluconate, an intermediate metabolite of the oxPPP, was significantly reduced by ∼50% in treated versus nontreated failing hearts, supporting the inhibitory effect of 6-aminonicotinamide. Our study indicates an important contribution of the oxPPP activity to cardiac oxidative stress in HF, which is particularly pronounced during common physiological changes such as postprandial glycemic peaks.

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Inhibition of triosephosphate isomerase by phosphoenolpyruvate in the feedback-regulation of glycolysis

Open Biology ◽

10.1098/rsob.130232 ◽

2014 ◽

Vol 4 (3) ◽

pp. 130232 ◽

Cited By ~ 47

Author(s):

Nana-Maria Grüning ◽

Dijun Du ◽

Markus A. Keller ◽

Ben F. Luisi ◽

Markus Ralser

Keyword(s):

Oxidative Stress ◽

Pyruvate Kinase ◽

Feedback Loop ◽

Triosephosphate Isomerase ◽

Feedback Regulation ◽

Pentose Phosphate ◽

Yeast Cells ◽

Dihydroxyacetone Phosphate

The inhibition of triosephosphate isomerase (TPI) in glycolysis by the pyruvate kinase (PK) substrate phosphoenolpyruvate (PEP) results in a newly discovered feedback loop that counters oxidative stress in cancer and actively respiring cells. The mechanism underlying this inhibition is illuminated by the co-crystal structure of TPI with bound PEP at 1.6 Å resolution, and by mutational studies guided by the crystallographic results. PEP is bound to the catalytic pocket of TPI and occludes substrate, which accounts for the observation that PEP competitively inhibits the interconversion of glyceraldehyde-3-phosphate and dihydroxyacetone phosphate. Replacing an isoleucine residue located in the catalytic pocket of TPI with valine or threonine altered binding of substrates and PEP, reducing TPI activity in vitro and in vivo . Confirming a TPI-mediated activation of the pentose phosphate pathway (PPP), transgenic yeast cells expressing these TPI mutations accumulate greater levels of PPP intermediates and have altered stress resistance, mimicking the activation of the PK–TPI feedback loop. These results support a model in which glycolytic regulation requires direct catalytic inhibition of TPI by the pyruvate kinase substrate PEP, mediating a protective metabolic self-reconfiguration of central metabolism under conditions of oxidative stress.

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In Vitro Study of the Toxicity Mechanisms of Nanoscale Zero-Valent Iron (nZVI) and Released Iron Ions Using Earthworm Cells

Nanomaterials ◽

10.3390/nano10112189 ◽

2020 ◽

Vol 10 (11) ◽

pp. 2189

Author(s):

Jaroslav Semerad ◽

Natividad Isabel Navarro Pacheco ◽

Alena Grasserova ◽

Petra Prochazkova ◽

Martin Pivokonsky ◽

...

Keyword(s):

Oxidative Stress ◽

Reactive Oxygen Species ◽

Reactive Oxygen ◽

Zero Valent Iron ◽

Oxygen Species ◽

Iron Ions ◽

Short Term ◽

Nanoscale Zero Valent Iron ◽

Toxicity Mechanisms

During the last two decades, nanomaterials based on nanoscale zero-valent iron (nZVI) have ranked among the most utilized remediation technologies for soil and groundwater cleanup. The high reduction capacity of elemental iron (Fe0) allows for the rapid and cost-efficient degradation or transformation of many organic and inorganic pollutants. Although worldwide real and pilot applications show promising results, the effects of nZVI on exposed living organisms are still not well explored. The majority of the recent studies examined toxicity to microbes and to a lesser extent to other organisms that could also be exposed to nZVI via nanoremediation applications. In this work, a novel approach using amoebocytes, the immune effector cells of the earthworm Eisenia andrei, was applied to study the toxicity mechanisms of nZVI. The toxicity of the dissolved iron released during exposure was studied to evaluate the effect of nZVI aging with regard to toxicity and to assess the true environmental risks. The impact of nZVI and associated iron ions was studied in vitro on the subcellular level using different toxicological approaches, such as short-term immunological responses and oxidative stress. The results revealed an increase in reactive oxygen species production following nZVI exposure, as well as a dose-dependent increase in lipid peroxidation. Programmed cell death (apoptosis) and necrosis were detected upon exposure to ferric and ferrous ions, although no lethal effects were observed at environmentally relevant nZVI concentrations. The decreased phagocytic activity further confirmed sublethal adverse effects, even after short-term exposure to ferric and ferrous iron. Detection of sublethal effects, including changes in oxidative stress-related markers such as reactive oxygen species and malondialdehyde production revealed that nZVI had minimal impacts on exposed earthworm cells. In comparison to other works, this study provides more details regarding the effects of the individual iron forms associated with nZVI aging and the cell toxicity effects on the specific earthworms’ immune cells that represent a suitable model for nanomaterial testing.

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Short-term exposure to positively charged polystyrene nanoparticles causes oxidative stress and membrane destruction in cyanobacteria

Environmental Science Nano ◽

10.1039/c9en00807a ◽

2019 ◽

Vol 6 (10) ◽

pp. 3072-3079 ◽

Cited By ~ 8

Author(s):

Li-Juan Feng ◽

Jian-Wei Li ◽

Elvis Genbo Xu ◽

Xiao-Dong Sun ◽

Fan-Ping Zhu ◽

...

Keyword(s):

Oxidative Stress ◽

Glutathione Metabolism ◽

Ecological Risks ◽

Hydrophobic Core ◽

Short Term ◽

Polystyrene Nanoparticles ◽

Plastic Debris ◽

Short Term Exposure ◽

Positively Charged ◽

Freshwater Environments

PS-NH2 exposure causes disruption of glutathione metabolism and damage to the membrane hydrophobic core of S. elongatus. This enhanced the understanding of the ecological risks of plastic debris to freshwater environments.

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Moderate Caloric Restriction Partially Improved Oxidative Stress Markers in Obese Humans

Antioxidants ◽

10.3390/antiox10071018 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1018

Author(s):

Dominika Kanikowska ◽

Alina Kanikowska ◽

Ewelina Swora-Cwynar ◽

Marian Grzymisławski ◽

Maki Sato ◽

...

Keyword(s):

Oxidative Stress ◽

Antioxidant Status ◽

Dietary Intervention ◽

Real Life ◽

Ros Generation ◽

Oxidative Stress Markers ◽

Short Term ◽

Anthropometric Parameters ◽

Stress Markers

Oxidative stress and inflammation are implicated in obesity. Therefore, we investigated whether moderate and short-term calorie restriction (CR) reflects a real-life situation, mediates weight loss, and improves oxidative stress markers. We analyzed oxidative stress markers in patients with obesity undergoing moderate CR. Serum oxidative stress markers (myeloperoxidase (MPO), superoxide dismutase (SOD), catalase, total antioxidant status (TAS), and reactive oxygen species (ROS) (generation by endothelial cells in vitro)) were measured in 53 subjects (mean BMI 37.8 ± 5.9 kg/m2) who underwent 8 weeks of CR, which included a reduction of 300–500 kcal/day. MPO was the most CR-sensitive parameter. The mean level of serum MPO in patients with obesity was 20% higher than that in post CR intervention (p < 0.001). SOD increased by 12% after CR (p < 0.05), which was largely due to the improvement in glucose tolerance and the reduction in insulin resistance after CR. Other tested parameters were not modified during the treatment. CR resulted in an expected decrease in body weight (by 5.9 ± 4.6 kg, p < 0.0001) and other anthropometric parameters. Additionally, it was accompanied by a significant change in hsCRP, hsTNF alpha, hsIL-6, leptin (all p < 0.0001), and HOMA-IR (p < 0.05). Cardiovascular and metabolic parameters were also partially improved. Short-term, moderate CR partially improves antioxidant capacity but is enough to substantially change anthropometric parameters in obese patients. Our observations indicate that mimicking real-life situations and low-cost dietary intervention can be successfully implemented in obesity treatment with a simultaneous moderate effect on antioxidant status.

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229 SHORT-TERM EXPOSURE OF MATURE OOCYTES TO A NITRIC OXIDE DONOR FOR INDUCING OXIDATIVE STRESS RESISTANCE ON IN VITRO-PRODUCED BOVINE EMBRYOS

Reproduction Fertility and Development ◽

10.1071/rdv27n1ab229 ◽

2015 ◽

Vol 27 (1) ◽

pp. 204

Author(s):

C. Cheuquemán ◽

P. Loren ◽

M. Arias ◽

J. Risopatrón ◽

R. Felmer ◽

...

Keyword(s):

Oxidative Stress ◽

Gene Expression ◽

Embryo Quality ◽

Nitric Oxide Donor ◽

Control Group ◽

Relative Gene Expression ◽

Short Term ◽

Short Term Exposure ◽

Relative Gene

Recent studies have shown that short-term exposure of oocytes to stressors such as hydrostatic pressure, osmotic stress, and oxidative stress might induce stress tolerance in embryos. In this research we studied the effect of short-term exposure of bovine in vitro-matured cumulus-oocyte complexes (COC) with a nitric oxide donor (SNP) on IVF, embryo development, embryo quality, and relative gene expression related to cell redox state regulation. The COC were selected and matured in TCM 199 supplemented with 10% inactivated FBS, 6 mg mL–1 of LH, 6 mg mL–1 of FSH, 1 mg mL–1 of oestradiol, and 0.2 mmol of pyruvate and then incubated for 22 to 24 h at 38.5°C in 5% CO2 in a humidified atmosphere (n = 12). Before IVF, mature COC were incubated during 1 h with different concentration of sodium nitroprusside, SNP (control without SNP, 10–6 M, 10–5 M, and 10–4 M SNP) in maturation media at 38.5°C and 5% CO2 in a humidified atmosphere. For IVF procedure, oocytes of each treatment and sperm of one bull were co-incubated for 18 to 20 h at 38.5°C and 5% CO2. Presumptive zygotes were separately cultured until Day 7 under mineral oil at 38.5°C and 5% CO2, 5%O2, and 90% N2 in a humidified atmosphere. Embryo quality was analysed by staining with CDX2 antibody for trophectoderm cells and compared with total embryo cells stained with Hoechst 33342. Relative gene expression for each treatment were evaluated after RNA extraction and cDNA synthesis in Stratagene MX 3000P real-time equipment with Agilent qPCR software MX pro 4.1 version. Differences between experimental groups (n = 12) were measured using a one-way ANOVA test in the STATGRAPHICS plus 5.1 version software. P < 0.05 was considered statistically significant. Cleavage percentage at 72 h post-insemination was significantly different between the control and 10–4 M SNP group (82 ± 8.4% v. 77 ± 7.1%, respectively) and between 10–5 M and 10–4 M SNP group (84.9 ± 4.1% v. 77 ± 7.1%, respectively). Blastocyst percentage at 7 days of culture was significantly different between control and 10–4 M SNP group (34.1 ± 7.8% v. 26.2 ± 4.9%, respectively). Embryo development between control group and treatments was similar within early, expanded, and hatched blastocyst percentage. Embryo quality of expanded blastocyst was similar between control group and treatments (ICM: TE). No significant differences in gene expression after SNP exposure was observed (iNOS, eNOS, nNOS, PRDX5, HSP70, HSP90, HIF1A, BCL2A). Oocytes incubated with a high concentration of SNP showed lower cleavage and blastocyst rates, showing that this treatment was deleterious for in vitro embryo production in bovine. However, there were no significant differences on embryo quality assessed by ICM : TE ratio and/or in gene expression pattern of 7-day cultured expanded blastocysts.

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