Physical developmental cues for the maturation of human pluripotent stem cell-derived cardiomyocytes

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are the most promising source of cardiomyocytes (CMs) for experimental and clinical applications, but their use is largely limited by a structurally and functionally immature phenotype that most closely resembles embryonic or fetal heart cells. The application of physical stimuli to influence hPSC-CMs through mechanical and bioelectrical transduction offers a powerful strategy for promoting more developmentally mature CMs. Here we summarize the major events associated with in vivo heart maturation and structural development. We then review the developmental state of in vitro derived hPSC-CMs, while focusing on physical (electrical and mechanical) stimuli and contributory (metabolic and hypertrophic) factors that are actively involved in structural and functional adaptations of hPSC-CMs. Finally, we highlight areas for possible future investigation that should provide a better understanding of how physical stimuli may promote in vitro development and lead to mechanistic insights. Advances in the use of physical stimuli to promote developmental maturation will be required to overcome current limitations and significantly advance research of hPSC-CMs for cardiac disease modeling, in vitro drug screening, cardiotoxicity analysis and therapeutic applications.published_or_final_versio

Cardiac Motion Analysis Based on Optical Flow on Real-Time Three-Dimensional Ultrasound Data

With relatively high frame rates and the ability to acquire volume data sets with a stationary transducer, 3D ultrasound systems, based on matrix phased array transducers, provide valuable three-dimensional information, from which quantitative measures of cardiac function can be extracted. Such analyses require segmentation and visual tracking of the left ventricular endocardial border. Due to the large size of the volumetric data sets, manual tracing of the endocardial border is tedious and impractical for clinical applications. Therefore the development of automatic methods for tracking three-dimensional endocardial motion is essential. In this study, we evaluate a four-dimensional optical flow motion tracking algorithm to determine its capability to follow the endocardial border in three dimensional ultrasound data through time. The four-dimensional optical flow method was implemented using three-dimensional correlation. We tested the algorithm on an experimental open-chest dog data set and a clinical data set acquired with a Philips' iE33 three-dimensional ultrasound machine. Initialized with left ventricular endocardial data points obtained from manual tracing at end-diastole, the algorithm automatically tracked these points frame by frame through the whole cardiac cycle. Finite element surfaces were fitted through the data points obtained by both optical flow tracking and manual tracing by an experienced observer for quantitative comparison of the results. Parameterization of the finite element surfaces was performed and maps displaying relative differences between the manual and semi-automatic methods were compared. The results showed good consistency with less than 10% difference between manual tracing and optical flow estimation on 73% of the entire surface. In addition, the optical flow motion tracking algorithm greatly reduced processing time (about 94% reduction compared to human involvement per cardiac cycle) for analyzing cardiac function in three-dimensional ultrasound data sets. A displacement field was computed from the optical flow output, and a framework for computation of dynamic cardiac information is introduced. The method was applied to a clinical data set from a heart transplant patient and dynamic measurements agreed with known physiology as well as experimental results

Geometric Phase, Curvature, and Extrapotentials in Constrained Quantum Systems

We derive an effective Hamiltonian for a quantum system constrained to a submanifold (the constraint manifold) of configuration space (the ambient space) by an infinite restoring force. We pay special attention to how this Hamiltonian depends on quantities which are external to the constraint manifold, such as the external curvature of the constraint manifold, the (Riemannian) curvature of the ambient space, and the constraining potential. In particular, we find the remarkable fact that the twisting of the constraining potential appears as a gauge potential in the constrained Hamiltonian. This gauge potential is an example of geometric phase, closely related to that originally discussed by Berry. The constrained Hamiltonian also contains an effective potential depending on the external curvature of the constraint manifold, the curvature of the ambient space, and the twisting of the constraining potential. The general nature of our analysis allows applications to a wide variety of problems, such as rigid molecules, the evolution of molecular systems along reaction paths, and quantum strip waveguides.Comment: 27 pages with 1 figure, submitted to Phys. Rev.

Evaluation of optical flow algorithms for tracking endocardial surfaces on three-dimensional ultrasound data

With relatively high frame rates and the ability to acquire volume data sets with a stationary transducer, 3D ultrasound systems, based on matrix phased array transducers, provide valuable three-dimensional information, from which quantitative measures of cardiac function can be extracted. Such analyses require segmentation and visual tracking of the left ventricular endocardial border. Due to the large size of the volumetric data sets, manual tracing of the endocardial border is tedious and impractical for clinical applications. Therefore the development of automatic methods for tracking three-dimensional endocardial motion is essential. In this study, we evaluate a four-dimensional optical flow motion tracking algorithm to determine its capability to follow the endocardial border in three dimensional ultrasound data through time. The four-dimensional optical flow method was implemented using three-dimensional correlation. We tested the algorithm on an experimental open-chest dog data set and a clinical data set acquired with a Philips' iE33 three-dimensional ultrasound machine. Initialized with left ventricular endocardial data points obtained from manual tracing at end-diastole, the algorithm automatically tracked these points frame by frame through the whole cardiac cycle.A finite element surface was fitted through the data points obtained by both optical flow tracking and manual tracing by an experienced observer for quantitative comparison of the results. Parameterization of the finite element surfaces was performed and maps displaying relative differences between the manual and semi-automatic methods were compared.The results showed good consistency between manual tracing and optical flow estimation on 73% of the entire surface with fewer than 10% difference. In addition, the optical flow motion tracking algorithm greatly reduced processing time (about 94% reduction compared to human involvement per cardiac cycle) for analyzing cardiac function in three-dimensional ultrasound data sets

Aircraft Conceptual Design Using Vehicle Sketch Pad

Vehicle Sketch Pad (VSP) is a parametric geometry modeling tool that is intended for use in the conceptual design of aircraft. The intent of this software is to rapidly model aircraft configurations without expending the expertise and time that is typically required for modeling with traditional Computer Aided Design (CAD) packages. VSP accomplishes this by using parametrically defined components, such as a wing that is defined by span, area, sweep, taper ratio, thickness to cord, and so on. During this phase of frequent design builds, changes to the model can be rapidly visualized along with the internal volumetric layout. Using this geometry-based approach, parameters such as wetted areas and cord lengths can be easily extracted for rapid external performance analyses, such as a parasite drag buildup. At the completion of the conceptual design phase, VSP can export its geometry to higher fidelity tools. This geometry tool was developed by NASA and is freely available to U.S. companies and universities. It has become integral to conceptual design in the Aeronautics Systems Analysis Branch (ASAB) here at NASA Langley Research Center and is currently being used at over 100 universities, aerospace companies, and other government agencies. This paper focuses on the use of VSP in recent NASA conceptual design studies to facilitate geometry-centered design methodology. Such a process is shown to promote greater levels of creativity, more rapid assessment of critical design issues, and improved ability to quickly interact with higher order analyses. A number of VSP vehicle model examples are compared to CAD-based conceptual design, from a designer perspective; comparisons are also made of the time and expertise required to build the geometry representations as well

Prospectus, November 27, 1985

https://spark.parkland.edu/prospectus_1985/1028/thumbnail.jp

Leptin fails to blunt the lipopolysaccharide-induced activation of the hypothalamic-pituitary-adrenal axis in rats

Copyright @ 2013 The authors. This work is licensed under a Creative Commons Attribution 3.0 Unported License.Obesity is a risk factor for sepsis morbidity and mortality, whereas the hypothalamic-pituitary-adrenal (HPA) axis plays a protective role in the body's defence against sepsis. Sepsis induces a profound systemic immune response and cytokines serve as excellent markers for sepsis as they act as mediators of the immune response. Evidence suggests that the adipokine leptin may play a pathogenic role in sepsis. Mouse endotoxaemic models present with elevated leptin levels and exogenously added leptin increased mortality whereas human septic patients have elevated circulating levels of the soluble leptin receptor (Ob-Re). Evidence suggests that leptin can inhibit the regulation of the HPA axis. Thus, leptin may suppress the HPA axis, impairing its protective role in sepsis.We hypothesised that leptin would attenuate the HPA axis response to sepsis.We investigated the direct effects of an i.p. injection of 2 mg/kg leptin on the HPA axis response to intraperitoneally injected 25 μg/kg lipopolysaccharide (LPS) in the male Wistar rat. We found that LPS potently activated the HPA axis, as shown by significantly increased plasma stress hormones, ACTH and corticosterone, and increased plasma interleukin 1β (IL1β) levels, 2 h after administration. Pre-treatment with leptin, 2 h before LPS administration, did not influence the HPA axis response to LPS. In turn, LPS did not affect plasma leptin levels. Our findings suggest that leptin does not influence HPA function or IL1b secretion in a rat model of LPS-induced sepsis, and thus that leptin is unlikely to be involved in the acute-phase endocrine response to bacterial infection in rats.The section is funded by grants from the MRC, BBSRC, NIHR and an Integrative Mammalian Biology (IMB) Capacity Building Award, and by a FP7-HEALTH-2009-241592 EuroCHIP grant and is supported by the NIHR Imperial Biomedical Research Centre Funding Scheme. This work is supported by a BBSRC Doctoral Training-Strategic Skills Award grant (BB/F017340/1)

Computational design of custom therapeutic cells to correct failing human cardiomyocytes

Background: Myocardial delivery of non-excitable cells—namely human mesenchymal stem cells (hMSCs) and c-kit+ cardiac interstitial cells (hCICs)—remains a promising approach for treating the failing heart. Recent empirical studies attempt to improve such therapies by genetically engineering cells to express specific ion channels, or by creating hybrid cells with combined channel expression. This study uses a computational modeling approach to test the hypothesis that custom hypothetical cells can be rationally designed to restore a healthy phenotype when coupled to human heart failure (HF) cardiomyocytes.Methods: Candidate custom cells were simulated with a combination of ion channels from non-excitable cells and healthy human cardiomyocytes (hCMs). Using a genetic algorithm-based optimization approach, candidate cells were accepted if a root mean square error (RMSE) of less than 50% relative to healthy hCM was achieved for both action potential and calcium transient waveforms for the cell-treated HF cardiomyocyte, normalized to the untreated HF cardiomyocyte.Results: Custom cells expressing only non-excitable ion channels were inadequate to restore a healthy cardiac phenotype when coupled to either fibrotic or non-fibrotic HF cardiomyocytes. In contrast, custom cells also expressing cardiac ion channels led to acceptable restoration of a healthy cardiomyocyte phenotype when coupled to fibrotic, but not non-fibrotic, HF cardiomyocytes. Incorporating the cardiomyocyte inward rectifier K+ channel was critical to accomplishing this phenotypic rescue while also improving single-cell action potential metrics associated with arrhythmias, namely resting membrane potential and action potential duration. The computational approach also provided insight into the rescue mechanisms, whereby heterocellular coupling enhanced cardiomyocyte L-type calcium current and promoted calcium-induced calcium release. Finally, as a therapeutically translatable strategy, we simulated delivery of hMSCs and hCICs genetically engineered to express the cardiomyocyte inward rectifier K+ channel, which decreased action potential and calcium transient RMSEs by at least 24% relative to control hMSCs and hCICs, with more favorable single-cell arrhythmia metrics.Conclusion: Computational modeling facilitates exploration of customizable engineered cell therapies. Optimized cells expressing cardiac ion channels restored healthy action potential and calcium handling phenotypes in fibrotic HF cardiomyocytes and improved single-cell arrhythmia metrics, warranting further experimental validation studies of the proposed custom therapeutic cells