Distinguishing cellular respiration vs. oxidative stress in turbid media using UV-excited autofluorescence spectroscopy

Reactive oxygen species (ROS), also known as free radicals, are generated during cellular respiration. Under normal conditions, the body has the ability to neutralize the effects of free radicals by using its antioxidant defenses. In the case of an imbalance between oxidants and antioxidants, free radical production exceeds the capacity of organic combustion, resulting in oxidative stress. Of all the cellular components compromised by the harmful effects of ROS, the cell membrane is the most severely affected owing to lipid peroxidation, which invariably leads to changes in the membrane structure and permeability. With lipid peroxidation of the cell membrane, some by-products can be detected and measured in tissues, blood, and other bodily fluids. The measurement of biomarkers of oxidative stress is commonly used to quantify lipid peroxidation of the cell membrane in humans, a species in which ROS can be considered as a cause or consequence of oxidative stress-related diseases. In dogs, few studies have demonstrated this correlation. The present review aims to identify current literature knowledge relating to oxidative stress diseases and their detection in dogs.

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Antioxidant Gene Expression in Vocal Hindbrain of a Teleost Fish

10.1101/354977 ◽

2018 ◽

Author(s):

Clara Liao ◽

Ni Y. Feng ◽

Andrew H. Bass

Keyword(s):

Oxidative Stress ◽

Breeding Season ◽

Motor Neurons ◽

Sound Production ◽

Cellular Respiration ◽

Type I ◽

Type Ii ◽

Antioxidant Genes ◽

Long Duration ◽

Significant Difference

ABSTRACTPlainfin midshipman fish (Porichthys notatus) have a remarkable capacity to generate long duration advertisement calls known as hums, each of which may last for close to two hours and be repeated throughout a night of courtship activity during the breeding season. The midshipman’s striking sound production capabilities provide a unique opportunity to investigate the mechanisms that motor neurons require for withstanding high-endurance activity. The temporal properties of midshipman vocal behaviors are largely controlled by a hindbrain central pattern generator that includes vocal motor neurons (VMN) that directly determine the activity pattern of target sonic muscles and, in turn, a sound’s pulse repetition rate, duration and pattern of amplitude modulation. Of the two adult midshipman male reproductive phenotypes -- types I and II-- only type I males acoustically court females with hums from nests that they build and guard, while type II males do not produce courtship hums but instead sneak or satellite spawn to steal fertilizations from type I males. A prior study using next generation RNA sequencing showed increased expression of a number of cellular respiration and antioxidant genes in the VMN of type I males during the breeding season, suggesting they help to combat potentially high levels of oxidative stress linked to this extreme behavior. This led to the question of whether the expression of these genes in the VMN would vary between actively humming versus non-humming states as well as between male morphs. Here, we tested the hypothesis that to combat oxidative stress, the VMN of reproductively active type I males would exhibit higher mRNA transcript levels for two superoxide dismutases (sod1,sod2) compared to the VMN of type II males and females that do not hum and in general both of which have a more limited vocal repertoire than type I males. The results showed no significant difference insod1transcript expression across reproductive morphs in the VMN and the surrounding hindbrain, and no difference ofsod2across the two male morphs and females in the SH. However, we observed a surprising, significantly lower expression ofsod2transcripts in the VMN of type I males as compared to type II males. We also found no significant difference insod1andsod2expression between actively humming and non-humming type I males in both the VMN and surrounding hindbrain. These findings overall lead us to conclude that increased transcription ofsod1andsod2is not necessary for combatting oxidative stress from the demands of the midshipman high-endurance vocalizations, but warrant future studies to assess protein levels, enzyme activity levels, as well as the expression of other antioxidant genes. These results also eliminate one of the proposed mechanisms that male midshipman use to combat potentially high levels of oxidative stress incurred by motor neurons driving long duration vocalization and provide more insight into how motor neurons are adapted to the performance of extreme behaviors.

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Low steady-state oxidative stress inhibits adipogenesis by altering mitochondrial dynamics and decreasing cellular respiration

Redox Biology ◽

10.1016/j.redox.2020.101507 ◽

2020 ◽

Vol 32 ◽

pp. 101507

Author(s):

Raquel Fernando ◽

Kristina Wardelmann ◽

Stefanie Deubel ◽

Richard Kehm ◽

Tobias Jung ◽

...

Keyword(s):

Oxidative Stress ◽

Steady State ◽

Mitochondrial Dynamics ◽

Cellular Respiration

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Inducible HSP70 regulates superoxide dismutase-2 and mitochondrial oxidative stress in the endothelial cells from developing lungs

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00264.2013 ◽

2014 ◽

Vol 306 (4) ◽

pp. L351-L360 ◽

Cited By ~ 28

Author(s):

Adeleye J. Afolayan ◽

Ru-Jeng Teng ◽

Annie Eis ◽

Ujala Rana ◽

Katarzyna A. Broniowska ◽

...

Keyword(s):

Oxidative Stress ◽

Endothelial Cells ◽

Superoxide Dismutase ◽

Specific Inhibitor ◽

Cellular Respiration ◽

Relaxation Response ◽

Mitochondrial Atpase ◽

Mitochondrial Oxidative Stress ◽

Atp Levels ◽

Superoxide Dismutase 2

Superoxide dismutase 2 (SOD-2) is synthesized in the cytosol and imported into the mitochondrial matrix, where it is activated and functions as the primary antioxidant for cellular respiration. The specific mechanisms that target SOD-2 to the mitochondria remain unclear. We hypothesize that inducible heat shock protein 70 (iHSP70) targets SOD-2 to the mitochondria via a mechanism facilitated by ATP, and this process is impaired in persistent pulmonary hypertension of the newborn (PPHN). We observed that iHSP70 interacts with SOD-2 and targets SOD-2 to the mitochondria. Interruption of iHSP70-SOD-2 interaction with 2-phenylethylenesulfonamide-μ (PFT-μ, a specific inhibitor of substrate binding to iHSP70 COOH terminus) and siRNA-mediated knockdown of iHSP70 expression disrupted SOD-2 transport to mitochondria. Increasing intracellular ATP levels by stimulation of respiration with CaCl2 facilitated the mitochondrial import of SOD-2, increased SOD-2 activity, and decreased the mitochondrial superoxide (O2·−) levels in PPHN pulmonary artery endothelial cells (PAEC) by promoting iHSP70-SOD-2 dissociation at the outer mitochondrial membrane. In contrast, oligomycin, an inhibitor of mitochondrial ATPase, decreased SOD-2 expression and activity and increased O2·− levels in the mitochondria of control PAEC. The basal ATP levels and degree of iHSP70-SOD-2 dissociation were lower in PPHN PAEC and lead to increased SOD-2 degradation in cytosol. In normal pulmonary arteries (PA), PFT-μ impaired the relaxation response of PA rings in response to nitric oxide (NO) donor, S-nitroso- N-acetyl-penicillamine. Pretreatment with Mito-Q, a mitochondrial targeted O2·− scavenger, restored the relaxation response in PA rings pretreated with PFT-μ. Our observations suggest that iHSP70 chaperones SOD-2 to the mitochondria. Impaired SOD-2-iHSP70 dissociation decreases SOD-2 import and contributes to mitochondrial oxidative stress in PPHN.

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Coenzyme Q10. Structure, role, sources and application in cosmetology

Aesthetic Cosmetology and Medicine ◽

10.52336/acm.2021.10.4.06 ◽

2021 ◽

Vol 10 (4) ◽

pp. 205-208

Author(s):

Agata Zejfer ◽

Keyword(s):

Oxidative Stress ◽

Amino Acids ◽

Coenzyme Q10 ◽

Citric Acid Cycle ◽

Krebs Cycle ◽

Eukaryotic Cell ◽

Cellular Respiration ◽

Healthy Skin ◽

Acid Cycle ◽

Cellular Energy

Cell division, the synthesis of amino acids, glycosaminoglycans and enzymes as well as the maturation of collagen and elastin fibers are the basic processes that take place in healthy skin. These reactions are possible due to participation of cellular energy of ATP (adenosine triphosphate), which is produced inside the mitochondria. Within one eukaryotic cell there are from several to several thousand mitochondria. The process of producing energy in the mitochondria is called cellular respiration, known also as the citric acid cycle or the Krebs cycle. Coenzyme Q10 is a key carrier of electrons in the respiratory chain. At the same time, it is responsible for the neutralization of free radicals formed in the Krebs cycle, leading to reduction oxidative stress and minimalization the aging processes of tissues. The article presents the collected information on coenzyme Q10, its structure, role, sources and application in cosmetology. Supplementation in case of coenzyme Q10 deficiency in the skin is possible through the use of appropriate cosmetic preparations, cosmetology treatments and dietary supplements.

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Oxidative Stress Induces Mitochondrial Compromise in CD4 T Cells From Chronically HCV-Infected Individuals

Frontiers in Immunology ◽

10.3389/fimmu.2021.760707 ◽

2021 ◽

Vol 12 ◽

Author(s):

Madison Schank ◽

Juan Zhao ◽

Ling Wang ◽

Lam Ngoc Thao Nguyen ◽

Dechao Cao ◽

...

Keyword(s):

Oxidative Stress ◽

T Cells ◽

Mitochondrial Dna ◽

Cd4 T Cells ◽

Hcv Infection ◽

Cellular Respiration ◽

Peroxisome Proliferator ◽

Mtdna Copy Number ◽

Mitochondrial Mass ◽

Mitochondrial Functions

We have previously shown that chronic Hepatitis C virus (HCV) infection can induce DNA damage and immune dysfunctions with excessive oxidative stress in T cells. Furthermore, evidence suggests that HCV contributes to increased susceptibility to metabolic disorders. However, the underlying mechanisms by which HCV infection impairs cellular metabolism in CD4 T cells remain unclear. In this study, we evaluated mitochondrial mass and intracellular and mitochondrial reactive oxygen species (ROS) production by flow cytometry, mitochondrial DNA (mtDNA) content by real-time qPCR, cellular respiration by seahorse analyzer, and dysregulated mitochondrial-localized proteins by Liquid Chromatography-Mass Spectrometry (LC-MS) in CD4 T cells from chronic HCV-infected individuals and health subjects. Mitochondrial mass was decreased while intracellular and mitochondrial ROS were increased, expressions of master mitochondrial regulators peroxisome proliferator-activated receptor 1 alpha (PGC-1α) and mitochondrial transcription factor A (mtTFA) were down-regulated, and oxidative stress was increased while mitochondrial DNA copy numbers were reduced. Importantly, CRISPR/Cas9-mediated knockdown of mtTFA impaired cellular respiration and reduced mtDNA copy number. Furthermore, proteins responsible for mediating oxidative stress, apoptosis, and mtDNA maintenance were significantly altered in HCV-CD4 T cells. These results indicate that mitochondrial functions are compromised in HCV-CD4 T cells, likely via the deregulation of several mitochondrial regulatory proteins.

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A Novel Insight Into the Fate of Cardiomyocytes in Ischemia-Reperfusion Injury: From Iron Metabolism to Ferroptosis

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.799499 ◽

2021 ◽

Vol 9 ◽

Author(s):

Jing-yan Li ◽

Shuang-qing Liu ◽

Ren-qi Yao ◽

Ying-ping Tian ◽

Yong-ming Yao

Keyword(s):

Oxidative Stress ◽

Reperfusion Injury ◽

Iron Homeostasis ◽

Ischemia Reperfusion Injury ◽

Therapeutic Option ◽

Ischemia Reperfusion ◽

Oxygen Metabolism ◽

Reperfusion Therapy ◽

Cellular Respiration ◽

Excess Iron

Ischemia-reperfusion injury (IRI), critically involved in the pathology of reperfusion therapy for myocardial infarction, is closely related to oxidative stress the inflammatory response, and disturbances in energy metabolism. Emerging evidence shows that metabolic imbalances of iron participate in the pathophysiological process of cardiomyocyte IRI [also termed as myocardial ischemia-reperfusion injury (MIRI)]. Iron is an essential mineral required for vital physiological functions, including cellular respiration, lipid and oxygen metabolism, and protein synthesis. Nevertheless, cardiomyocyte homeostasis and viability are inclined to be jeopardized by iron-induced toxicity under pathological conditions, which is defined as ferroptosis. Upon the occurrence of IRI, excessive iron is transported into cells that drive cardiomyocytes more vulnerable to ferroptosis by the accumulation of reactive oxygen species (ROS) through Fenton reaction and Haber–Weiss reaction. The increased ROS production in ferroptosis correspondingly leads cardiomyocytes to become more sensitive to oxidative stress under the exposure of excess iron. Therefore, ferroptosis might play an important role in the pathogenic progression of MIRI, and precisely targeting ferroptosis mechanisms may be a promising therapeutic option to revert myocardial remodeling. Notably, targeting inhibitors are expected to prevent MIRI deterioration by suppressing cardiomyocyte ferroptosis. Here, we review the pathophysiological alterations from iron homeostasis to ferroptosis together with potential pathways regarding ferroptosis secondary to cardiovascular IRI. We also provide a comprehensive analysis of ferroptosis inhibitors and initiators, as well as regulatory genes involved in the setting of MIRI.

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Novel Antioxidant Therapy with the Immediate Precursor to Glutathione, γ-Glutamylcysteine (GGC), Ameliorates LPS-Induced Cellular Stress in In Vitro 3D-Differentiated Airway Model from Primary Cystic Fibrosis Human Bronchial Cells

Antioxidants ◽

10.3390/antiox9121204 ◽

2020 ◽

Vol 9 (12) ◽

pp. 1204

Author(s):

Chris K. Hewson ◽

Alexander Capraro ◽

Sharon L. Wong ◽

Elvis Pandzic ◽

Ling Zhong ◽

...

Keyword(s):

Oxidative Stress ◽

Cystic Fibrosis ◽

Stress Responses ◽

Therapeutic Potential ◽

Membrane Integrity ◽

The Body ◽

Stress Granule ◽

Cellular Respiration ◽

Granule Formation ◽

Lps Challenge

Systemic glutathione deficiency, inflammation, and oxidative stress are hallmarks of cystic fibrosis (CF), an inherited disease that causes persistent lung infections and severe damage to the respiratory system and many of the body organs. Improvements to current antioxidant therapeutic strategies are needed. The dietary supplement, γ-glutamylcysteine (GGC), which is the immediate precursor to glutathione, rapidly boosts cellular glutathione levels following a single dose in healthy individuals. Efficacy of GGC against oxidative stress induced by Pseudomonas aeruginosa, which is a common and chronic pathogen infecting lungs of CF patients, remains unassessed. Primary mucocilliary differentiated airway (bronchial and/or nasal) epithelial cells were created from four individuals with CF. Airway oxidative stress and inflammation was induced by P. aeruginosa lipopolysaccharide (LPS). Parameters including global proteomics alterations, cell redox state (glutathione, oxidative stress), pro-inflammatory mediators (IL-8, IDO-1), and cellular health (membrane integrity, stress granule formation, cell metabolic viability) were assayed under six experimental conditions: (1) Mock, (2) LPS-challenged (3) therapeutic, (4) prophylactic (5) therapeutic and prophylactic and (6) GGC alone. Proteomic analysis identified perturbation of several pathways related to cellular respiration and stress responses upon LPS challenge. Most of these were resolved when cells were treated with GGC. While GGC did not resolve LPS-induced IL-8 and IDO-1 activity, it effectively attenuated LPS-induced oxidative stress and stress granule formation, while significantly increasing total intracellular glutathione levels, metabolic viability and improving epithelial cell barrier integrity. Both therapeutic and prophylactic treatments were successful. Together, these findings indicate that GGC has therapeutic potential for treatment and prevention of oxidative stress-related damage to airways in cystic fibrosis.

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Novel antioxidant therapy with the immediate precursor to glutathione, γ-glutamylcysteine (GGC), ameliorates LPS-induced cellular stress in an in vitro cystic fibrosis model

10.1101/2020.05.27.119990 ◽

2020 ◽

Author(s):

Chris K Hewson ◽

Alexander Capraro ◽

Sharon L Wong ◽

Elvis Pandzic ◽

Bentotage S.M Fernando ◽

...

Keyword(s):

Oxidative Stress ◽

Cystic Fibrosis ◽

Cell Viability ◽

Stress Responses ◽

Therapeutic Potential ◽

Membrane Integrity ◽

Stress Granule ◽

Cellular Respiration ◽

Granule Formation ◽

Lps Challenge

AbstractIntroductionGlutathione deficiency and chronic bacterial inflammation exacerbates the oxidative stress damage to airways in cystic fibrosis. Improvements to current antioxidant therapeutic strategies are needed. Dietary supplement, γ-glutamylcysteine (GGC), the immediate precursor to glutathione, rapidly boosts cellular glutathione levels following a single dose in healthy individuals. Efficacy of GGC against Pseudomonas aeruginosa derived lipopolysaccharide (LPS), a prominent factor in mediating both bacterial virulence and host responses, in CF remains unassessed.MethodsPrimary F508del/F508del mucociliary differentiated bronchial and nasal epithelial cells were created to model LPS-induced oxidative stress and inflammation of CF. The proteomic signature of GGC treated cells was resolved by qLC-MS/MS. Parameters including cell redox state (glutathione, ROS), anti-inflammatory mediators (IL-8, IDO-1) and cellular health (membrane integrity, stress granule formation and cell viability) were assayed.ResultsProteomic analysis identified perturbation of several pathways related to cellular respiration and stress responses upon LPS challenge. Most of these were resolved when cells were treated with GGC. While GGC did not resolve LPS-induced IL-8 and IDO-1 activity, it effectively attenuated LPS-induced ROS and stress granule formation, while significantly increasing intracellular glutathione levels and improving epithelial cell barrier integrity. Moreover, we compared the effect of GGC with thiols NAC and glutathione on cell viability. GGC was the only thiol that increased cell viability; protecting cells against LPS induced cell death. Both therapeutic and prophylactic treatments were successful.ConclusionTogether, these findings indicate that GGC has therapeutic potential for treatment and prevention of oxidative stress related damage to airways in Cystic Fibrosis.

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Transcriptome-wide Gene Expression Analysis in Peri-implantitis Reveals Candidate Cellular Pathways

JDR Clinical & Translational Research ◽

10.1177/23800844211045297 ◽

2021 ◽

pp. 238008442110452

Author(s):

A. Martin ◽

P. Zhou ◽

B.B. Singh ◽

G.A. Kotsakis

Keyword(s):

Oxidative Stress ◽

Gene Expression ◽

Soft Tissue ◽

Actin Polymerization ◽

Inflammatory Responses ◽

Degradation Products ◽

Rna Extraction ◽

Orthopedic Implants ◽

Oral Bacteria ◽

Cellular Respiration

Objective: Peri-implantitis is a condition resulting in destructive inflammation in the peri-implant soft tissue barrier. Clinically, it demonstrates vast clinical differences to periodontitis that suggest distinct inflammatory mechanisms. Implant-derived titanium particles (i-TiPs) frequently found around diseased implants appear to alter the microenvironment and confer resistance to antibiotic treatments. Studies in orthopedic implants have demonstrated potent inflammatory responses to i-TiPs involving a variety of cell types in aseptic conditions. Nonetheless, the genetic programs of cells surveilling and supporting the peri-implant soft tissue barrier in response to the combined challenges of biomaterial degradation products and oral bacteria are poorly defined. Thus, we studied gene expression specific to oral peri-implant inflammatory disease. Methods: Peri-implant tissues were collected from healthy or diseased implants (N = 10) according to the 2018 classification criteria. Following RNA extraction and purification, a gene-level view of the transcriptome was obtained via a next-generation transcriptome-wide microarray profiling workflow (Clariom S; Applied Biosystems) that covers >20,000 well-annotated genes. A discovery analysis assessed global differential expression of genes and identified pathways in peri-implant health versus disease. Results: Genes involved in the endosomal-lysosomal pathway, such as actin polymerization, were strongly upregulated in diseased tissues (P < .05), proposing increased intracellular activities in response to bacteria and i-TiPs. Cellular respiration pathways involved in oxidative stress were highly transcribed in all peri-implant samples, suggesting that implant-specific factors may trigger a constant state of oxidative stress. Conclusion: Within the limitations of this discovery study, expressive upregulation of genes in the endosomal-lysosomal and oxidative stress pathway suggests that inflammation related to receptor-driven responses to extracellular signals, such as i-TiPs and pathogens, may have a crucial role in peri-implantitis. Results warrant external replication in validation cohorts. Knowledge Transfer Statement: Our findings regarding physiologic processes affected by peri-implantitis could advance knowledge of the mechanisms and consequences of the disease. Understanding the cellular programs that partake in peri-implant inflammation has the potential to translate to novel treatment strategies for patients with peri-implantitis.

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