Measuring Bioenergetic Signatures of Peripheral Nerve Segments by Extracellular Flux Analysis

2020 ◽  
pp. 191-203 ◽  
Author(s):  
Bogdan Beirowski
Keyword(s):  
Peripheral Nerve ◽  
Flux Analysis ◽  
Extracellular Flux

scholarly journals Prominent Indomethacin-Induced Enteropathy in Fcgriib Defi-cient lupus Mice: An Impact of Macrophage Responses and Immune Deposition in Gut

2021 ◽  
Vol 22 (3) ◽  
pp. 1377
Author(s):  
Thansita Bhunyakarnjanarat ◽  
Kanyarat Udompornpitak ◽  
Wilasinee Saisorn ◽  
Bhumdhanin Chantraprapawat ◽  
Peerapat Visitchanakun ◽  
...  
Keyword(s):  
Cytokine Production ◽  
High Dose ◽  
Flux Analysis ◽  
Wild Type ◽  
Fc Gamma Receptor ◽  
Serum Cytokines ◽  
Extracellular Flux ◽  
Gamma Receptor ◽  
Gut Leakage ◽  
Gastrointestinal Permeability

A high dose of NSAIDs, a common analgesic, might induce lupus activity through several NSAIDs adverse effects including gastrointestinal permeability defect (gut leakage) and endotoxemia. Indomethacin (25 mg/day) was orally administered for 7 days in 24-wk-old Fc gamma receptor IIb deficient (FcgRIIb-/-) mice, an asymptomatic lupus model (increased anti-dsDNA without lupus nephritis), and age-matched wild-type (WT) mice. Severity of indomethacin-induced enteropathy in FcgRIIb-/- mice was higher than WT mice as demonstrated by survival analysis, intestinal injury (histology, immune-deposition, and intestinal cytokines), gut leakage (FITC-dextran assay and endotoxemia), serum cytokines, and lupus characteristics (anti-dsDNA, renal injury, and proteinuria). Prominent responses of FcgRIIb-/- macrophages toward lipopolysaccharide (LPS) compared to WT cells due to the expression of only activating-FcgRs without inhibitory-FcgRIIb were demonstrated. Extracellular flux analysis indicated the greater mitochondria activity (increased respiratory capacity and respiratory reserve) in FcgRIIb-/- macrophages with a concordant decrease in glycolysis activity when compared to WT cells. In conclusion, gut leakage-induced endotoxemia is more severe in indomethacin-administered FcgRIIb-/- mice than WT, possibly due to the enhanced indomethacin toxicity from lupus-induced intestinal immune-deposition. Due to a lack of inhibitory-FcgRIIb expression, mitochondrial function, and cytokine production of FcgRIIb-/- macrophages were more prominent than WT cells. Hence, lupus disease-activation from NSAIDs-enteropathy-induced gut leakage is possible.

009 Self-Reported Sleep Efficiency and Duration are Associated with Systemic Bioenergetic Function in Community-Dwelling Adults

SLEEP ◽  
2021 ◽  
Vol 44 (Supplement_2) ◽  
pp. A4-A4
Author(s):  
H Matthew Lehrer ◽  
Lauren Chu ◽  
Martica Hall ◽  
Kyle Murdock
Keyword(s):  
Sleep Duration ◽  
Mononuclear Cells ◽  
Basic Research ◽  
Basal Respiration ◽  
Community Dwelling ◽  
Sleep Efficiency ◽  
Flux Analysis ◽  
Spare Capacity ◽  
Peripheral Blood Mononuclear ◽  
Extracellular Flux

Abstract Introduction Sleep is important for aging, health, and disease, but its cellular role in these outcomes is poorly understood. Basic research suggests that disturbed and insufficient sleep impair mitochondrial bioenergetics, which is involved in numerous aging-related chronic conditions. However, the relationship between sleep and bioenergetics has not been examined in humans. We examined associations of self-reported sleep with systemic bioenergetic function in peripheral blood mononuclear cells (PBMCs) of community-dwelling adults. Methods N = 43 adults (79% female) ages 48–70 (M = 61.63, SD = 5.99) completed the Pittsburgh Sleep Quality Index (PSQI) from which key components of sleep (satisfaction, alertness, timing, efficiency, and duration) were calculated. Participants provided blood samples from which PBMCs were isolated and measured for bioenergetics using extracellular flux analysis. Associations of sleep components with bioenergetic parameters, including the Bioenergetic Health Index (BHI), were examined. Results In bivariate analyses, lower sleep efficiency was associated with lower maximal respiration, spare capacity, and BHI (ps < 0.05). Longer sleep duration was associated with lower BHI (p < 0.01) and later sleep timing was associated with higher basal respiration, ATP-linked respiration, maximal respiration, spare capacity, and non-mitochondrial respiration (ps < 0.05). After adjustment for age, sex, and body mass index, lower sleep efficiency (β = 0.52, p < 0.01) and longer sleep duration (β = -0.43, p < 0.01) were associated with lower BHI. Conclusion Self-reported indices of sleep efficiency and duration are related to systemic bioenergetic function in humans, suggesting a possible cellular pathway linking sleep to health. Support (if any) T32HL082610

Extracellular Flux Analysis to Investigate the Impact of NF-κB on in Colorectal Carcinoma (CRC)

2021 ◽  
pp. 293-303
Author(s):  
Daria Capece ◽  
Daniela Verzella ◽  
Federica Begalli ◽  
Jason Bennett ◽  
Daniel D’Andrea ◽  
...  
Keyword(s):  
Colorectal Carcinoma ◽  
Flux Analysis ◽  
Extracellular Flux ◽  
The Impact

scholarly journals Application of extracellular flux analysis for determining mitochondrial function in mammalian oocytes and early embryos

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Bethany Muller ◽  
Niamh Lewis ◽  
Tope Adeniyi ◽  
Henry J. Leese ◽  
Daniel R. Brison ◽  
...  
Keyword(s):  
Real Time ◽  
Oocyte Maturation ◽  
Small Groups ◽  
Mitochondrial Function ◽  
Mitochondrial Activity ◽  
Flux Analysis ◽  
Mammalian Oocyte ◽  
Extracellular Flux ◽  
Early Embryos ◽  
The Impact

AbstractMitochondria provide the major source of ATP for mammalian oocyte maturation and early embryo development. Oxygen Consumption Rate (OCR) is an established measure of mitochondrial function. OCR by mammalian oocytes and embryos has generally been restricted to overall uptake and detailed understanding of the components of OCR dedicated to specific molecular events remains lacking. Here, extracellular flux analysis (EFA) was applied to small groups of bovine, equine, mouse and human oocytes and bovine early embryos to measure OCR and its components. Using EFA, we report the changes in mitochondrial activity during the processes of oocyte maturation, fertilisation, and pre-implantation development to blastocyst stage in response to physiological demands in mammalian embryos. Crucially, we describe the real time partitioning of overall OCR to spare capacity, proton leak, non-mitochondrial and coupled respiration – showing that while activity changes over the course of development in response to physiological demand, the overall efficiency is unchanged. EFA is shown to be able to measure mitochondrial function in small groups of mammalian oocytes and embryos in a manner which is robust, rapid and easy to use. EFA is non-invasive and allows real-time determination of the impact of compounds on OCR, facilitating an assessment of the components of mitochondrial activity. This provides proof-of-concept for EFA as an accessible system with which to study mammalian oocyte and embryo metabolism.

scholarly journals Increased Susceptibility to Ethylmercury-Induced Mitochondrial Dysfunction in a Subset of Autism Lymphoblastoid Cell Lines

2015 ◽  
Vol 2015 ◽  
pp. 1-13 ◽  
Author(s):  
Shannon Rose ◽  
Rebecca Wynne ◽  
Richard E. Frye ◽  
Stepan Melnyk ◽  
S. Jill James
Keyword(s):  
Oxidative Stress ◽  
Mitochondrial Dysfunction ◽  
Cell Lines ◽  
Autism Spectrum ◽  
Reserve Capacity ◽  
Flux Analysis ◽  
Lymphoblastoid Cell Lines ◽  
Extracellular Flux ◽  
Respiratory Parameters ◽  
Increased Risk

The association of autism spectrum disorders with oxidative stress, redox imbalance, and mitochondrial dysfunction has become increasingly recognized. In this study, extracellular flux analysis was used to compare mitochondrial respiration in lymphoblastoid cell lines (LCLs) from individuals with autism and unaffected controls exposed to ethylmercury, an environmental toxin known to deplete glutathione and induce oxidative stress and mitochondrial dysfunction. We also tested whether pretreating the autism LCLs with N-acetyl cysteine (NAC) to increase glutathione concentrations conferred protection from ethylmercury. Examination of 16 autism/control LCL pairs revealed that a subgroup (31%) of autism LCLs exhibited a greater reduction in ATP-linked respiration, maximal respiratory capacity, and reserve capacity when exposed to ethylmercury, compared to control LCLs. These respiratory parameters were significantly elevated at baseline in the ethylmercury-sensitive autism subgroup as compared to control LCLs. NAC pretreatment of the sensitive subgroup reduced (normalized) baseline respiratory parameters and blunted the exaggerated ethylmercury-induced reserve capacity depletion. These findings suggest that the epidemiological link between environmental mercury exposure and an increased risk of developing autism may be mediated through mitochondrial dysfunction and support the notion that a subset of individuals with autism may be vulnerable to environmental influences with detrimental effects on development through mitochondrial dysfunction.

scholarly journals The profiles of mitochondrial respiration and glycolysis using extracellular flux analysis in porcine enterocyte IPEC-J2

2015 ◽  
Vol 1 (3) ◽  
pp. 239-243 ◽  
Author(s):  
Bie Tan ◽  
Hao Xiao ◽  
Fengna Li ◽  
Liming Zeng ◽  
Yulong Yin
Keyword(s):  
Mitochondrial Respiration ◽  
Flux Analysis ◽  
Extracellular Flux

scholarly journals Mitochondrial Respiration in Female Zucker Rats: Effects of Obesity and Short-Term Metformin Treatment

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 1682-1682
Author(s):  
Shannon Rose ◽  
Eugenia Carvalho ◽  
David Irby ◽  
Sirish Bennuri ◽  
Alexandria Beebe ◽  
...  
Keyword(s):  
Mitochondrial Respiration ◽  
Coupling Efficiency ◽  
Zucker Rats ◽  
Proton Leak ◽  
Acid Oxidation ◽  
Flux Analysis ◽  
Metformin Treatment ◽  
Short Term ◽  
Funding Sources ◽  
Extracellular Flux

Abstract Objectives As childhood obesity rates climb, so has the incidence of type 2 diabetes among children and adolescents. Metformin, an FDA approved anti-hyperglycemic drug, is thought to target the mitochondria, but its effects on mitochondrial function in obesity have not been well studied. We used an obese Zucker rat model to investigate the effects of obesity and short-term metformin treatment on mitochondrial respiration. Methods Five-week old female Zucker rats (n = 16 lean, n = 16 obese) were fed AIN-93 G diet for 8 weeks before equally randomized to receive metformin (mixed in diet at 1 g/kg of feed); thus forming 4 groups with 8 rats each: lean +/– metformin and obese +/– metformin. Rats were sacrificed 10 weeks post-metformin treatment and spleens, perigonadal visceral adipose tissue (VAT) and skeletal muscle (SM; gracilis) collected. Mitochondrial respiration was measured in splenocytes by extracellular flux analysis and in VAT and SM fibers by high resolution respirometry. Results Effects of obesity on mitochondrial respiration were found in VAT and SM, but not splenocytes. In VAT, obese rats exhibited increased OXPHOS capacity over lean rats when octanoylcarnitine and malate were provided as substrates (obese vs lean: 1.33 vs 0.76 pmol O2/s/mg; SEdiff = 0.18, P = .005), and after subsequent additions of pyruvate (P = .012), glutamate (P = .009), and succinate (P = .045). In SM, OXPHOS capacity was increased when octanoylcarnitine and malate were provided as substrates (obese vs lean: 12.18 vs 5.45 pmol O2/s/mg; SEdiff = 2.31, P = .011) in obese vs lean rats. Metformin effects were observed only in splenocytes: coupling efficiency was decreased (metformin vs no metformin; 56.2% vs 69.8%; SEdiff = 4.1%, P = 0.005) and proton leak was increased (P < .001) in metformin treated rats as compared to rats not treated with metformin. Conclusions We found obesity was associated with increased mitochondrial respiration, particularly fatty acid oxidation, in VAT and SM. Short-term metformin treatment did not alter mitochondrial respiration in VAT or SM, but was found to increase proton leak and reduce coupling efficiency in splenocytes. Funding Sources Arkansas Children's Research Institute, Arkansas Biosciences Institute (R.H). S.R. and E.C. are also supported by the National Institute of General Medical Sciences of the National Institutes of Health.

scholarly journals Lactate Reprograms Energy and Lipid Metabolism in Glucose-Deprived Oxidative Glioma Stem Cells

Metabolites ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 325
Author(s):  
Noriaki Minami ◽  
Kazuhiro Tanaka ◽  
Takashi Sasayama ◽  
Eiji Kohmura ◽  
Hideyuki Saya ◽  
...  
Keyword(s):  
Stem Cells ◽  
Fatty Acid Biosynthesis ◽  
Malignant Tumors ◽  
Tricarboxylic Acid Cycle ◽  
Carbon Sources ◽  
Glucose Deprivation ◽  
Metabolic Reprogramming ◽  
Glioma Stem Cells ◽  
Flux Analysis ◽  
Extracellular Flux

Fast-growing tumors satisfy their bioenergetic needs by supplementing glucose with alternative carbon sources. Cancer stem cells are the most versatile and robust cells within malignant tumors. They avoid potentially lethal metabolic and other types of stress through flexible reprogramming of relevant pathways, but it has remained unclear whether alternative carbon sources are important for the maintenance of their tumor-propagating ability. Here we assessed the ability of glycolytic and oxidative murine glioma stem cells (GSCs) to grow in an ultralow glucose medium. Sphere formation assays revealed that exogenous lactate and acetate reversed the growth impairment of oxidative GSCs in such medium. Extracellular flux analysis showed that lactate supported oxygen consumption in these cells, whereas metabolomics analysis revealed that it increased the intracellular levels of tricarboxylic acid cycle intermediates, ATP, and GTP as well as increased adenylate and guanylate charge. Lactate also reversed the depletion of choline apparent in the glucose-deprived cells as well as reprogrammed phospholipid and fatty acid biosynthesis. This metabolic reprogramming was associated with a more aggressive phenotype of intracranial tumors formed by lactate-treated GSCs. Our results thus suggest that lactate is an important alternative energetic and biosynthetic substrate for oxidative GSCs, and that it sustains their growth under conditions of glucose deprivation.

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