Enhanced Assessment of Oxidative Tissue Damage
Abstract
Breathing 100% oxygen at an atmospheric pressure > 1 atmosphere absolute (ATA) is hyperbaric oxygen (HBO2). It is indicated for trea,ting air or gas embolism, decompression sickness, carbon monoxide poisoning, delayed radiation injury, and several other complicatio,ns. HBO2 is also used in diving operations to prevent nitrogen gas bubble formation and reduce decompression time and risk of illnes,s. HBO2 is, however, toxic to the central nervous system (CNS) and lungs, and possibly to other tissues, limiting its potential appl,ication in clinical medicine and diving. Although this has been known for more than a century, the precise mechanisms remain unknown,. The Duke Center for Hyperbaric Medicine (CHMEP) has been supported by the Office of Naval Research for over 20 years to determine,how HBO2 causes CNS and pulmonary toxicity and to test pharmacological mitigation strategies. The overall goal of the CHMEPs resear,ch is to safely extend the HBO2 dosage (depth time) and improve diver performance. Research completed by the CHMEPs team, and oth,ers, suggest HBO2 impacts mitochondrial fusion and fission (dynamics), the selective clearance of dysfunctional or damaged mitochond,ria (mitophagy), and synthesis of new mitochondria (biogenesis). The processes are tightly regulated and comprise the mitochondrial,quality control cycle, and are linked to mitochondrial function. It may be that the increase in oxidants produced by mitochondria in, HBO2 trigger mitochondrial fission and mitophagy and this compromises cellular bioenergetics. To test this hypothesis, the CHMEP re,searchers rely on established laboratory techniques, e.g. immunoblotting, real-time quantitative reverse transcription polymerase ch,ain reaction, and respirometry. Respirometry is the benchmark for assessing mitochondrial dysfunction. The purpose of this proposal,is to enhance the CHMEPs ability to assess oxidative tissue damage in current and future DOD supported research. To accomplish this,, we propose to usethe LI-COR Odyssey Fc protein and nucleic acid imager, Hansatech Oxytherm+ respirometry system and Miltenyi Biot,ec gentleMACS Dissociator. The LI-COR Odyssey Fc protein and nucleic acid imager uses one optical density to ensure protein band re,plication between measurements, automatically finds lanes and subtracts background, eliminates saturation, and can measure chemilumi,nescent or fluorescent labeled proteins. The Hansatech Oxytherm+ respirometry system and Miltenyi Biotec gentleMACS Dissociator will, reduce mitochondrial isolation time and improve measurement throughput. Taken together, these equipment will serve to further our u,nderstanding of how HBO2 affects cellular function, and help guide the development of safe and effective prevention and treatment st,rategies that will improve operational capabilities. Moreover, the equipment will enhance education and training opportunities for s,tudents pursuing or considering careers in STEM.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Apr 01, 2022
- Source ID
- N000142212383
Entities
People
- Heath G Gasier
Organizations
- Duke University
- Office of Naval Research
- United States Navy