Biomedical Engineering and Bioengineering Commons^™

Hyperoxia Causes Mitochondrial Fragmentation In Pulmonary Endothelial Cells By Increasing Expression Of Pro-Fission Proteins, Cui Ma, Andreas M. Beyer, Matthew J. Durand, Anne V. Clough, Daling Zhu, Laura Norwood Toro, Maia Terashvili, Johnathan D. Ebben, R. Blake Hill, Said H. Audi, Meetha Medhora, Elizabeth R. Jacobs

Biomedical Engineering Faculty Research and Publications

Objective—We explored mechanisms that alter mitochondrial structure and function in pulmonary endothelial cells (PEC) function after hyperoxia.

Approach and Results—Mitochondrial structures of PECs exposed to hyperoxia or normoxia were visualized and mitochondrial fragmentation quantified. Expression of pro-fission or fusion proteins or autophagy-related proteins were assessed by Western blot. Mitochondrial oxidative state was determined using mito-roGFP. Tetramethylrhodamine methyl ester estimated mitochondrial polarization in treatment groups. The role of mitochondrially derived reactive oxygen species in mt-fragmentation was investigated with mito-TEMPOL and mitochondrial DNA (mtDNA) damage studied by using ENDO III (mt-tat-endonuclease III), a protein that repairs mDNA damage. Drp-1 (dynamin-related …

Mg2+ Differentially Regulates Two Modes Of Mitochondrial Ca2+ Uptake In Isolated Cardiac Mitochondria: Implications For Mitochondrial Ca2+ Sequestration, Christoph A. Blomeyer, Jason Bazil, David F. Stowe, Ranjan K. Dash, Amadou Ks Camara

Biomedical Engineering Faculty Research and Publications

The manner in which mitochondria take up and store Ca²⁺ remains highly debated. Recent experimental and computational evidence has suggested the presence of at least two modes of Ca²⁺ uptake and a complex Ca²⁺ sequestration mechanism in mitochondria. But how Mg²⁺ regulates these different modes of Ca²⁺ uptake as well as mitochondrial Ca²⁺ sequestration is not known. In this study, we investigated two different ways by which mitochondria take up and sequester Ca²⁺ by using two different protocols. Isolated guinea pig cardiac mitochondria were exposed to varying concentrations of CaCl₂ in the …

Safety And Efficacy Of Ranolazine For The Treatment Of Chronic Angina Pectoris, Mohammed Aldakkak, David F. Stowe, Amadou K.S. Camara

Biomedical Engineering Faculty Research and Publications

Coronary heart disease is a global malady and it is the leading cause of death in the United States. Chronic stable angina is the most common manifestation of coronary heart disease and it results from the imbalance between myocardial oxygen supply and demand due to reduction in coronary blood flow. Therefore, in addition to lifestyle changes, commonly used pharmaceutical treatments for angina (nitrates, ß-blockers, Ca2+ channel blockers) are aimed at increasing blood flow or decreasing O2 demand. However, patients may continue to experience symptoms of angina. Ranolazine is a relatively new drug with anti-anginal and anti-arrhythmic effects. Its anti-anginal mechanism …

Tyrosine Nitration Of Voltage-Dependent Anion Channels In Cardiac Ischemia-Reperfusion: Reduction By Peroxynitrite Scavenging, Meiying Yang, Amadou K.S. Camara, Bassam T. Wakim, Yifan Zhou, Ashish K. Gadicherla, Wai-Meng Kwok, David F. Stowe

Biomedical Engineering Faculty Research and Publications

Excess superoxide (O₂⁻) and nitric oxide (NO) forms peroxynitrite (ONOO⁻) during cardiac ischemia reperfusion (IR) injury, which in turn induces protein tyrosine nitration (tyr-N). Mitochondria are both a source of and target for ONOO⁻. Our aim was to identify specific mitochondrial proteins that display enhanced tyr-N after cardiac IR injury, and to explore whether inhibiting O₂⁻/ONOO⁻ during IR decreases mitochondrial protein tyr-N and consequently improves cardiac function. We show here that IR increased tyr-N of 35 and 15 kDa mitochondrial proteins using Western blot analysis with 3-nitrotyrosine antibody. Immunoprecipitation …

Damage To Mitochondrial Complex I During Cardiac Ischemia Reperfusion Injury Is Reduced Indirectly By Anti-Anginal Drug Ranolazine, Ashish K. Gadicherla, David F. Stowe, William E. Antholine, Meiying Yang, Amadou K.S. Camara

Biomedical Engineering Faculty Research and Publications

Ranolazine, an anti-anginal drug, is a late Na⁺ channel current blocker that is also believed to attenuate fatty acid oxidation and mitochondrial respiratory complex I activity, especially during ischemia. In this study, we investigated if ranolazine's protective effect against cardiac ischemia/reperfusion (IR) injury is mediated at the mitochondrial level and specifically if respiratory complex I (NADH Ubiquinone oxidoreductase) function is protected. We treated isolated and perfused guinea pig hearts with ranolazine just before 30 min ischemia and then isolated cardiac mitochondria at the end of 30 min ischemia and/or 30 min ischemia followed by 10 min reperfusion. We utilized …

Enhanced Charge-Independent Mitochondrial Free Ca2+ And Attenuated Adp-Induced Nadh Oxidation By Isoflurane: Implications For Cardioprotection, Bhawana Agarwal, Amadou K.S. Camara, David F. Stowe, Zeljko J. Bosnjak, Ranjan K. Dash

Biomedical Engineering Faculty Research and Publications

Modulation of mitochondrial free Ca^{2 +} ([Ca^{2 +}]_m) is implicated as one of the possible upstream factors that initiates anesthetic-mediated cardioprotection against ischemia–reperfusion (IR) injury. To unravel possible mechanisms by which volatile anesthetics modulate [Ca^{2 +}]_m and mitochondrial bioenergetics, with implications for cardioprotection, experiments were conducted to spectrofluorometrically measure concentration-dependent effects of isoflurane (0.5, 1, 1.5, 2 mM) on the magnitudes and time-courses of [Ca^{2 +}]_m and mitochondrial redox state (NADH), membrane potential (ΔΨ_m), respiration, and matrix volume. Isolated mitochondria from rat hearts were energized with 10 …

Mitochondrial Approaches To Protect Against Cardiac Ischemia And Reperfusion Injury, Amadou K.S. Camara, Martin Bienengraeber, David F. Stowe

Biomedical Engineering Faculty Research and Publications

The mitochondrion is a vital component in cellular energy metabolism and intracellular signaling processes. Mitochondria are involved in a myriad of complex signaling cascades regulating cell death vs. survival. Importantly, mitochondrial dysfunction and the resulting oxidative and nitrosative stress are central in the pathogenesis of numerous human maladies including cardiovascular diseases, neurodegenerative diseases, diabetes, and retinal diseases, many of which are related. This review will examine the emerging understanding of the role of mitochondria in the etiology and progression of cardiovascular diseases and will explore potential therapeutic benefits of targeting the organelle in attenuating the disease process. Indeed, recent advances …

Enhanced Na+/H+ Exchange During Ischemia And Reperfusion Impairs Mitochondrial Bioenergetics And Myocardial Function, Mohammed Aldakkak, David F. Stowe, James S. Heisner, Marisha Spence, Amadou K.S. Camara

Biomedical Engineering Faculty Research and Publications

Inhibition of Na+/H+ exchange (NHE) during ischemia reduces cardiac injury due to reduced reverse mode Na+/Ca2+ exchange. We hypothesized that activating NHE-1 at buffer pH 8 during ischemia increases mitochondrial oxidation, Ca2+ overload, and reactive O2 species (ROS) levels and worsens functional recovery in isolated hearts and that NHE inhibition reverses these effects. Guinea pig hearts were perfused with buffer at pH 7.4 (control) or pH 8 +/- NHE inhibitor eniporide for 10 minutes before and for 10 minutes after 35- minute ischemia and then for 110 minutes with pH 7.4 buffer alone. Mitochondrial NADH and FAD, [Ca2+], and superoxide …

Katp Channel Openers Have Opposite Effects On Mitochondrial Respiration Under Different Energetic Conditions, Matthias L. Riess, Amadou K.S. Camara, André Heinen, Janis T. Eells, Michele M. Henry, David F. Stowe

Biomedical Engineering Faculty Research and Publications

Mitochondrial (m) K_ATP channel opening has been implicated in triggering cardiac preconditioning. Its consequence on mitochondrial respiration, however, remains unclear. We investigated the effects of two different K_ATP channel openers and antagonists on mitochondrial respiration under two different energetic conditions. Oxygen consumption was measured for complex I (pyruvate/malate) or complex II (succinate with rotenone) substrates in mitochondria from fresh guinea pig hearts. One of two mK_ATP channel openers, pinacidil or diazoxide, was given before adenosine diphosphate in the absence or presence of an mK_ATP channel antagonist, glibenclamide or 5-hydroxydecanoate. Without ATP synthase inhibition, both mK_{ATP …}