Quantitative Assessment of Mitochondrial Morphology and Electrophysiological Function in the Diabetic Heart.

Mitochondria within a cardiomyocyte form a highly dynamic network that undergoes fusion and fission events in response to acute and chronic stressors, such as hyperglycemia and diabetes mellitus. Changes in mitochondrial architecture and morphology not only reflect their capacity for oxidative phosphorylation and ATP synthesis but also impact their subcellular localization and interaction with other organelles. The role of these ultrastructural abnormalities in modulating electrophysiological properties and excitation-contraction coupling remains largely unknown and warrants direct investigation considering the growing appreciation of the functional and structural coupling between the mitochondrial network, the calcium cycling machinery, and sarcolemmal ion channels in the cardiac myocyte. In this Methods in Molecular Biology chapter, we provide a protocol that allows for a quantitative assessment of mitochondrial shape and morphology in control and diabetic hearts that had undergone detailed electrophysiological measurements using high resolution optical action potential (AP) mapping.

A hyper-viscoelastic uniaxial characterization of collagenous embolus analogs in acute ischemic stroke.

PURPOSE: Acute ischemic stroke is a leading cause of death and morbidity worldwide. Despite advances in medical technology, nearly 30% of strokes result in incomplete vessel recanalization. Recent studies have demonstrated that clot composition correlates with success rates of mechanical thrombectomy procedures. To understand clot behavior during thrombectomy, which exerts considerable strains on thrombi, in vitro studies must characterize the rate-dependent high-strain behavior of embolus analogs (EAs) with different formation conditions, which can be used to fit models of hyper-viscoelasticity. METHODS: In this study, the effect of collagen infiltration as a carotid-induced collagen-rich thrombosis surrogate is considered as a contributor to embolus analog high-strain stiffness, when compared to 40% hematocrit EAs. RESULTS: EA high-strain stiffnesses, characterized on a uniaxial load frame, increase by an order of magnitude for collagenous clot analogs. Chandler loop analogs show high-strain stiffnesses and clot compositions commensurate with previous reports of stroke patient clots, and collagenous clots show significant increase in stiffness when compared to stroke patient clots. Finally, hyper-viscoelastic curve fitting demonstrates the asymmetry between tension and compression. Nonlinear, rate-dependent models that consider clot-stiffening behavior match the high strain stiffness of clots fairly well. Furthermore, we demonstrate that the stability of the elastic energy needs to be considered to obtain optimal curve fits for high-strain, rate dependent data. CONCLUSION: This study provides a framework for the development of dynamically formed EAs that mimic the mechanical and structural properties of in vivo clots and provides parameters for numerical simulation of clot behavior with hyper-viscoelastic models.