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https://www.readbyqxmd.com/read/27889640/inhibition-of-autophagy-with-bafilomycin-and-chloroquine-decreases-mitochondrial-quality-and-bioenergetic-function-in-primary-neurons
#1
Matthew Redmann, Gloria A Benavides, Taylor F Berryhill, Willayat Y Wani, Xiaosen Ouyang, Michelle S Johnson, Saranya Ravi, Stephen Barnes, Victor M Darley-Usmar, Jianhua Zhang
Autophagy is an important cell recycling program responsible for the clearance of damaged or long-lived proteins and organelles. Pharmacological modulators of this pathway have been extensively utilized in a wide range of basic research and pre-clinical studies. Bafilomycin A1 and chloroquine are commonly used compounds that inhibit autophagy by targeting the lysosomes but through distinct mechanisms. Since it is now clear that mitochondrial quality control, particularly in neurons, is dependent on autophagy, it is important to determine whether these compounds modify cellular bioenergetics...
November 18, 2016: Redox Biology
https://www.readbyqxmd.com/read/27872984/cinnamic-acid-derivatives-enhance-the-efficacy-of-transarterial-embolization-in-a-rat-model-of-hepatocellular-carcinoma
#2
Luke R Wilkins, David L Brautigan, Hanping Wu, Hooman Yarmohammadi, Ewa Kubicka, Vlad Serbulea, Norbert Leitinger, Wendy Liu, John R Haaga
INTRODUCTION: We hypothesize that the combination of transarterial embolization (TAE) plus inhibition of lactate export will limit anaerobic metabolism and reduce tumor survival compared to TAE alone. The purpose of this study was to test this hypothesis in a rat model of hepatocellular carcinoma (HCC). METHODS: Rat N1-S1 hepatoma cells were assayed in vitro using the Seahorse XF analyzer to measure extracellular acidification (lactate excretion) comparing effects of the addition of caffeic acid (CA) or ferulic acid (FA) or UK-5099 with control...
November 21, 2016: Cardiovascular and Interventional Radiology
https://www.readbyqxmd.com/read/27791989/stimulation-of-the-toll-like-receptor-3-promotes-metabolic-reprogramming-in-head-and-neck-carcinoma-cells
#3
Mathieu Veyrat, Sylvère Durand, Marion Classe, Tanja Matijevic Glavan, Natalie Oker, Nikiforos-Ioannis Kapetanakis, Xiaojun Jiang, Aurore Gelin, Philippe Herman, Odile Casiraghi, David Zagzag, David Enot, Pierre Busson, Benjamin Vérillaud
In this study, a possible link between the innate immune recognition receptor TLR3 and metabolic reprogramming in Head and Neck carcinoma (HNC) cells was investigated. The effects of TLR3 stimulation/knock-down were assessed under several culture conditions in 4 HNC cell-lines by cell growth assays, targeted metabolomics, and glycolysis assays based on time-resolved analysis of proton release (Seahorse analyzer). The stimulation of TLR3 by its synthetic agonist Poly(A:U) resulted in a faster growth of HNC cells under low foetal calf serum conditions...
October 25, 2016: Oncotarget
https://www.readbyqxmd.com/read/27784195/mitochondrial-spare-respiratory-capacity-is-negatively-correlated-with-nuclear-reprogramming-efficiency
#4
Yan Zhou, Rasha Abdelkadhem Al-Saaidi, Paula Fernandez-Guerra, Kristine K Freude, Rikke Katrine Jentoft Olsen, Uffe Birk Jensen, Niels Gregersen, Poul Hyttel, Lars Bolund, Lars Aagaard, Peter Bross, Yonglun Luo
Nuclear reprogramming efficiency has been shown to be highly variable among different types of somatic cells and different individuals, yet the underlying mechanism remains largely unknown. Several studies have shown that reprogramming of fibroblasts into induced pluripotent stem cells (iPSCs) requires remodeling of mitochondria and a metabolic shift from an oxidative state to a more glycolytic state. In this study, we evaluated the nuclear reprogramming efficiency in relation to mitochondrial bioenergetic parameters of fibroblasts from seven different human individuals...
November 28, 2016: Stem Cells and Development
https://www.readbyqxmd.com/read/27764131/generating-rho-0-cells-using-mesenchymal-stem-cell-lines
#5
Mercedes Fernández-Moreno, Tamara Hermida-Gómez, M Esther Gallardo, Andrea Dalmao-Fernández, Ignacio Rego-Pérez, Rafael Garesse, Francisco J Blanco
INTRODUCTION: The generation of Rho-0 cells requires the use of an immortalization process, or tumor cell selection, followed by culture in the presence of ethidium bromide (EtBr), incurring the drawbacks its use entails. The purpose of this work was to generate Rho-0 cells using human mesenchymal stem cells (hMSCs) with reagents having the ability to remove mitochondrial DNA (mtDNA) more safely than by using EtBr. METHODOLOGY: Two immortalized hMSC lines (3a6 and KP) were used; 143B...
2016: PloS One
https://www.readbyqxmd.com/read/27741233/extracellular-redox-regulation-of-intracellular-reactive-oxygen-generation-mitochondrial-function-and-lipid-turnover-in-cultured-human-adipocytes
#6
Albert R Jones Iv, Tova Meshulam, Marcus F Oliveira, Nathan Burritt, Barbara E Corkey
BACKGROUND: Many tissues play an important role in metabolic homeostasis and the development of diabetes and obesity. We hypothesized that the circulating redox metabolome is a master metabolic regulatory system that impacts all organs and modulates reactive oxygen species (ROS) production, lipid peroxidation, energy production and changes in lipid turnover in many cells including adipocytes. METHODS: Differentiated human preadipocytes were exposed to the redox couples, lactate (L) and pyruvate (P), β-hydroxybutyrate (βOHB) and acetoacetate (Acoc), and the thiol-disulfides cysteine/ cystine (Cys/CySS) and GSH/GSSG for 1...
2016: PloS One
https://www.readbyqxmd.com/read/27718558/a-novel-methodology-for-bioenergetic-analysis-of-plasmodium-falciparum-reveals-a-glucose-regulated-metabolic-shift-and-enables-mode-of-action-analyses-of-mitochondrial-inhibitors
#7
Tomoyo Sakata-Kato, Dyann F Wirth
Given that resistance to all drugs in clinical use has arisen, discovery of new anti-malarial drug targets is eagerly anticipated. The Plasmodium mitochondrion has been considered a promising drug target largely based on its significant divergence from the host organelle as well as its involvement in ATP production and pyrimidine biosynthesis. However, the functions of Plasmodium mitochondrial protein complexes and associated metabolic pathways are not fully characterized. Here, we report the development of novel and robust bioenergetic assay protocols for Plasmodium falciparum asexual parasites utilizing a Seahorse Bioscience XFe24 Extracellular Flux Analyzer...
October 9, 2016: ACS Infectious Diseases
https://www.readbyqxmd.com/read/27710853/hyperglycemia-induced-damage-to-mitochondrial-respiration-in-renal-mesangial-and-tubular-cells-implications-for-diabetic-nephropathy
#8
Anna Czajka, Afshan N Malik
Damage to renal tubular and mesangial cells is central to the development of diabetic nephropathy (DN), a complication of diabetes which can lead to renal failure. Mitochondria are the site of cellular respiration and produce energy in the form of ATP via oxidative phosphorylation, and mitochondrial dysfunction has been implicated in DN. Since the kidney is an organ with high bioenergetic needs, we postulated that hyperglycemia causes damage to renal mitochondria resulting in bioenergetic deficit. The bioenergetic profiles and the effect of hyperglycemia on cellular respiration of human primary mesangial (HMCs) and proximal tubular cells (HK-2) were compared in normoglycemic and hyperglycemic conditions using the seahorse bio-analyzer...
September 17, 2016: Redox Biology
https://www.readbyqxmd.com/read/27702678/parental-care-improves-immunity-in-the-seahorse-hippocampus-erectus
#9
Tingting Lin, Dong Zhang, Xin Liu, Dongxue Xiao
In the present study, the sexual dimorphism in immune response in the seahorse Hippocampus erectus in which males compete for mates and invest heavily in parental care was assessed. Variability in immunocompetence in virginal seahorses with differing levels of sexual maturity (i.e., immaturity, early maturity and maturity) and with different mating statuses (i.e., virginal, experienced mating failure and experienced mating success) were analyzed by evaluating immune parameters in the plasma. Additionally, ultrastructural characteristics of the inner epithelium of the brood pouch were compared between males that had experienced mating failure and those that had succeeded...
October 1, 2016: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/27698021/macrophages-promote-oxidative-metabolism-to-drive-nitric-oxide-generation-in-response-to-trypanosoma-cruzi
#10
Sue-Jie Koo, Imran H Chowdhury, Bartosz Szczesny, Xianxiu Wan, Nisha J Garg
Trypanosoma cruzi is the causative agent of chronic chagasic cardiomyopathy. Why macrophages (mφs), the early responders to infection, fail to achieve parasite clearance is not known. Mouse (RAW 264.7) and human (THP-1 and primary) mφs were infected for 3 h and 18 h with T. cruzi TcI isolates, SylvioX10/4 (SYL, virulent) and TCC (nonpathogenic), which represent mφ stimulation and infection states, respectively. Mφs incubated with lipopolysaccharide and gamma interferon (LPS/IFN-γ) and with interleukin-4 (IL-4) were used as controls...
December 2016: Infection and Immunity
https://www.readbyqxmd.com/read/27604740/advanced-and-high-throughput-method-for-mitochondrial-bioenergetics-evaluation-in-neurotrauma
#11
Jignesh D Pandya, Patrick G Sullivan, Lai Yee Leung, Frank C Tortella, Deborah A Shear, Ying Deng-Bryant
Mitochondrial dysfunction is one of the key posttraumatic neuropathological events observed in various experimental models of traumatic brain injury (TBI). The extent of mitochondrial dysfunction has been associated with the severity and time course of secondary injury following brain trauma. Critically, several mitochondrial targeting preclinical drugs used in experimental TBI models have shown improved mitochondrial bioenergetics, together with cortical tissue sparing and cognitive behavioral outcome. Mitochondria, being a central regulator of cellular metabolic pathways and energy producer of cells, are of a great interest for researchers aiming to adopt cutting-edge methodology for mitochondrial bioenergetics assessment...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27586662/copper-zinc-superoxide-dismutase-cuznsod-an-antioxidant-gene-from-seahorse-hippocampus-abdominalis-molecular-cloning-sequence-characterization-antioxidant-activity-and-potential-peroxidation-function-of-its-recombinant-protein
#12
N C N Perera, G I Godahewa, Jehee Lee
Copper-zinc-superoxide dismutase (CuZnSOD) from Hippocampus abdominalis (HaCuZnSOD) is a metalloenzyme which belongs to the ubiquitous family of SODs. Here, we determined the characteristic structural features of HaCuZnSOD, analyzed its evolutionary relationships, and identified its potential immune responses and biological functions in relation to antioxidant defense mechanisms in the seahorse. The gene had a 5' untranslated region (UTR) of 67 bp, a coding sequence of 465 bp and a 3' UTR of 313 bp. The putative peptide consists of 154 amino acids...
October 2016: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/27545309/glucose-lactate-and-glutamine-but-not-glutamate-support-depolarization-induced-increased-respiration-in-isolated-nerve-terminals
#13
Michaela C Hohnholt, Vibe H Andersen, Lasse K Bak, Helle S Waagepetersen
Synaptosomes prepared from various aged and gene modified experimental animals constitute a valuable model system to study pre-synaptic mechanisms. Synaptosomes were isolated from whole brain and the XFe96 extracellular flux analyzer (Seahorse Bioscience) was used to study mitochondrial respiration and glycolytic rate in presence of different substrates. Mitochondrial function was tested by sequentially exposure of the synaptosomes to the ATP synthase inhibitor, oligomycin, the uncoupler FCCP (carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone) and the electron transport chain inhibitors rotenone and antimycin A...
August 22, 2016: Neurochemical Research
https://www.readbyqxmd.com/read/27500267/characterizing-the-mechanistic-pathways-of-the-instant-blood-mediated-inflammatory-reaction-in-xenogeneic-neonatal-islet-cell-transplantation
#14
David Liuwantara, Yi Vee Chew, Emmanuel J Favaloro, Joanne M Hawkes, Heather L Burns, Philip J O'Connell, Wayne J Hawthorne
INTRODUCTION: The instant blood-mediated inflammatory reaction (IBMIR) causes major loss of islets after transplantation and consequently represents the initial barrier to survival of porcine neonatal islet cell clusters (NICC) after xenotransplantation. METHODS: This study used novel assays designed to characterize the various immunologic components responsible for xenogeneic IBMIR to identify initiators and investigate processes of IBMIR-associated coagulation, complement activation and neutrophil infiltration...
June 2016: Transplantation Direct
https://www.readbyqxmd.com/read/27494181/identification-of-kca3-1-channel-as-a-novel-regulator-of-oxidative-phosphorylation-in-a-subset-of-pancreatic-carcinoma-cell-lines
#15
Ilya Kovalenko, Andrea Glasauer, Laura Schöckel, Daniel R P Sauter, Alexander Ehrmann, Florian Sohler, Andrea Hägebarth, Ivana Novak, Sven Christian
Pancreatic ductal adenocarcinoma (PDAC) represents the most common form of pancreatic cancer with rising incidence in developing countries and overall 5-year survival rates of less than 5%. The most frequent mutations in PDAC are gain-of-function mutations in KRAS as well as loss-of-function mutations in p53. Both mutations have severe impacts on the metabolism of tumor cells. Many of these metabolic changes are mediated by transporters or channels that regulate the exchange of metabolites and ions between the intracellular compartment and the tumor microenvironment...
2016: PloS One
https://www.readbyqxmd.com/read/27486856/differential-expression-of-complement-markers-in-normal-and-amd-transmitochondrial-cybrids
#16
Sonali Nashine, Marilyn Chwa, Mina Kazemian, Kunal Thaker, Stephanie Lu, Anthony Nesburn, Baruch D Kuppermann, M Cristina Kenney
PURPOSE: Variations in mitochondrial DNA (mtDNA) and abnormalities in the complement pathways have been implicated in the pathogenesis of age-related macular degeneration (AMD). This study was designed to determine the effects of mtDNA from AMD subjects on the complement pathway. METHODS: Transmitochondrial cybrids were prepared by fusing platelets from AMD and age-matched Normal subjects with Rho0 (lacking mtDNA) human ARPE-19 cells. Quantitative PCR and Western blotting were performed to examine gene and protein expression profiles, respectively, of complement markers in these cybrids...
2016: PloS One
https://www.readbyqxmd.com/read/27456956/gentisic-acid-sodium-salt-a-phenolic-compound-is-superior-to-norepinephrine-in-reversing-cardiovascular-collapse-hepatic-mitochondrial-dysfunction-and-lactic-acidemia-in-pseudomonas-aeruginosa-septic-shock-in-dogs
#17
Steven Mink, Subir K Roy Chowdhury, Jose Gotes, Zhao-Qin Cheng, Krika Kasian, Paul Fernyhough
BACKGROUND: The development of lactic acidemia (LA) in septic shock (SS) is associated with an ominous prognosis. We previously showed that the mechanism of LA in SS may relate to impaired hepatic uptake of lactate, but the mechanism was not clear. Uptake of lactate by the liver occurs by a membrane-associated, pH-dependent, antiport system known as the monocarboxylate transporter. In the hepatocyte, lactate can then be metabolized by oxidative phosphorylation or converted to glucose in the cytosol...
December 2016: Intensive Care Medicine Experimental
https://www.readbyqxmd.com/read/27445101/mitochondrial-oxygen-metabolism-in-primary-human-lens-epithelial-cells-association-with-age-diabetes-and-glaucoma
#18
M Kubota, Y B Shui, M Liu, F Bai, A J Huang, N Ma, D C Beebe, C J Siegfried
PURPOSE: The hypoxic environment around the lens is important for maintaining lens transparency. Lens epithelial cells (LECs) play a key role in lens metabolism. We measured oxygen consumption to assess the role of human LECs in maintaining hypoxia around the lens, as well as the impact of systemic and ocular diagnosis on these cells. METHODS: Baseline cellular respiration was measured in rabbit LECs (NN1003A), canine kidney epithelial cells (MDCK), trabecular meshwork cells (TM-5), and bovine corneal endothelial cells (CCEE) using a XF96 Extracellular Flux Analyzer (Seahorse Bioscience, North Billerica, MA), which measures oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) in vitro...
August 2016: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/27271893/mitochondrial-stress-tests-using-seahorse-respirometry-on-intact-dictyostelium-discoideum-cells
#19
Sui Lay, Oana Sanislav, Sarah J Annesley, Paul R Fisher
Mitochondria not only play a critical and central role in providing metabolic energy to the cell but are also integral to the other cellular processes such as modulation of various signaling pathways. These pathways affect many aspects of cell physiology, including cell movement, growth, division, differentiation, and death. Mitochondrial dysfunction which affects mitochondrial bioenergetics and causes oxidative phosphorylation defects can thus lead to altered cellular physiology and manifest in disease. The assessment of the mitochondrial bioenergetics can thus provide valuable insights into the physiological state, and the alterations to the state of the cells...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27231905/role-of-mitochondrial-dna-copy-number-alteration-in-human-renal-cell-carcinoma
#20
Chen-Sung Lin, Hui-Ting Lee, Ming-Huei Lee, Siao-Cian Pan, Chen-Yeh Ke, Allen Wen-Hsiang Chiu, Yau-Huei Wei
We investigated the role of mitochondrial DNA (mtDNA) copy number alteration in human renal cell carcinoma (RCC). The mtDNA copy numbers of paired cancer and non-cancer parts from five resected RCC kidneys after radical nephrectomy were determined by quantitative polymerase chain reaction (Q-PCR). An RCC cell line, 786-O, was infected by lentiviral particles to knock down mitochondrial transcriptional factor A (TFAM). Null target (NT) and TFAM-knockdown (TFAM-KD) represented the control and knockdown 786-O clones, respectively...
2016: International Journal of Molecular Sciences
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