Isolation and Mechanical Measurements of Myofibrils from Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.

Summary: Tension production and contractile properties are poorly characterized aspects of excitation-contraction coupling of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Previous approaches have been limited due to the small size and structural immaturity of early-stage hiPSC-CMs. We developed a substrate nanopatterning approach to produce hiPSC-CMs in culture with adult-like dimensions, T-tubule-like structures, and aligned myofibrils. We then isolated myofibrils from hiPSC-CMs and measured the tension and kinetics of activation and relaxation using a custom-built apparatus with fast solution switching. The contractile properties and ultrastructure of myofibrils more closely resembled human fetal myofibrils of similar gestational age than adult preparations. We also demonstrated the ability to study the development of contractile dysfunction of myofibrils from a patient-derived hiPSC-CM cell line carrying the familial cardiomyopathy MYH7 mutation (E848G). These methods can bring new insights to understanding cardiomyocyte maturation and developmental mechanical dysfunction of hiPSC-CMs with cardiomyopathic mutations. : In this article, Pioner and colleagues reported contractile properties of isolated myofibrils from hiPSC-CMs with highly mature morphology. This approach permits quantitative assessment of maturation and contractile properties of hiPSC-CMs and can be used to study the development of contractile dysfunction in genetically based cardiac diseases. The authors present a patient-derived cell line carrying a novel familial cardiomyopathy MYH7 mutation (E848G)

A Whole Cell Assay to Measure Caspase-6 Activity by Detecting Cleavage of Lamin A/C

Caspase-6 is a cysteinyl protease implicated in neurodegenerative conditions including Alzheimer's and Huntington's disease making it an attractive target for therapeutic intervention. A greater understanding of the role of caspase-6 in disease has been hampered by a lack of suitable cellular assays capable of specifically detecting caspase-6 activity in an intact cell environment. This is mainly due to the use of commercially available peptide substrates and inhibitors which lack the required specificity to facilitate development of this type of assay. We report here a 384-well whole-cell chemiluminescent ELISA assay that monitors the proteolytic degradation of endogenously expressed lamin A/C during the early stages of caspase-dependent apoptosis. The specificity of lamin A/C proteolysis by caspase-6 was demonstrated against recombinant caspase family members and further confirmed in genetic deletion studies. In the assay, plasma membrane integrity remained intact as assessed by release of lactate dehydrogenase from the intracellular environment and the exclusion of cell impermeable peptide inhibitors, despite the induction of an apoptotic state. The method described here is a robust tool to support drug discovery efforts targeting caspase-6 and is the first reported to specifically monitor endogenous caspase-6 activity in a cellular context

Non-unitary Evolution of Quantum Logics

In this work we present a dynamical approach to quantum logics. By changing the standard formalism of quantum mechanics to allow non-Hermitian operators as generators of time evolution, we address the question of how can logics evolve in time. In this way, we describe formally how a non-Boolean algebra may become a Boolean one under certain conditions. We present some simple models which illustrate this transition and develop a new quantum logical formalism based in complex spectral resolutions, a notion that we introduce in order to cope with the temporal aspect of the logical structure of quantum theory

Rapid transgenerational acclimation of a tropical reef fish to climate change

Understanding the capacity of species to acclimate and adapt to expected temperature increases is critical for making predictions about the biological impacts of global warming, yet it is one of the least certain aspects of climate change science. Tropical species are considered to be especially sensitive to climate change because they live close to their thermal maximum and exhibit limited capacity for acclimation. Here, we demonstrate that a tropical reef fish is highly sensitive to small increases in water temperature, but can rapidly acclimate over multiple generations. Acute exposure to elevated temperatures (+1.5 °C and +3.0 °C) predicted to occur this century caused a 15% and 30% respective decrease in individual's maximum ability to perform aerobic activities such as swimming or foraging, known as aerobic scope. However, complete compensation in aerobic scope occurred when both parents and offspring were reared throughout their lives at elevated temperature. Such acclimation could reduce the impact of warming temperatures and allow populations to persist across their current range. This study reveals the importance of transgenerational acclimation as a mechanism for coping with rapid climate change and highlights that single generation studies risk underestimating the potential of species to cope. © 2011 Macmillan Publishers Limited. All rights reserved