Mitochondrial connexin43 and mitochondrial KATP channels modulate triggered arrhythmias in mouse ventricular muscle.

Connexin43 (Cx43) exits as hemichannels in the inner mitochondrial membrane. We examined how mitochondrial Cx43 and mitochondrial KATP channels affect the occurrence of triggered arrhythmias. To generate cardiac-specific Cx43-deficient (cCx43-/-) mice, Cx43flox/flox mice were crossed with α-MHC (Myh6)-cre+/- mice. The resulting offspring, Cx43flox/flox/Myh6-cre+/- mice (cCx43-/- mice) and their littermates (cCx43+/+ mice), were used. Trabeculae were dissected from the right ventricles of mouse hearts. Cardiomyocytes were enzymatically isolated from the ventricles of mouse hearts. Force was measured with a strain gauge in trabeculae (22°C). To assess arrhythmia susceptibility, the minimal extracellular Ca2+ concentration ([Ca2+]o,min), at which arrhythmias were induced by electrical stimulation, was determined in trabeculae. ROS production was estimated with 2',7'-dichlorofluorescein (DCF), mitochondrial membrane potential with tetramethylrhodamine methyl ester (TMRM), and Ca2+ spark frequency with fluo-4 and confocal microscopy in cardiomyocytes. ROS production within the mitochondria was estimated with MitoSoxRed and mitochondrial Ca2+ with rhod-2 in trabeculae. Diazoxide was used to activate mitochondrial KATP. Most of cCx43-/- mice died suddenly within 8 weeks. Cx43 was present in the inner mitochondrial membrane in cCx43+/+ mice but not in cCx43-/- mice. In cCx43-/- mice, the [Ca2+]o,min was lower, and Ca2+ spark frequency, the slope of DCF fluorescence intensity, MitoSoxRed fluorescence, and rhod-2 fluorescence were higher. TMRM fluorescence was more decreased in cCx43-/- mice. Most of these changes were suppressed by diazoxide. In addition, in cCx43-/- mice, antioxidant peptide SS-31 and N-acetyl-L-cysteine increased the [Ca2+]o,min. These results suggest that Cx43 deficiency activates Ca2+ leak from the SR, probably due to depolarization of mitochondrial membrane potential, an increase in mitochondrial Ca2+, and an increase in ROS production, thereby causing triggered arrhythmias, and that Cx43 hemichannel deficiency may be compensated by activation of mitochondrial KATP channels in mouse hearts.

Improving the strength and toughness of macroscale double networks by exploiting Poisson's ratio mismatch.

We propose a new concept that utilizes the difference in Poisson's ratio between component materials as a strengthening mechanism that increases the effectiveness of the sacrificial bond toughening mechanism in macroscale double-network (Macro-DN) materials. These Macro-DN composites consist of a macroscopic skeleton imbedded within a soft elastic matrix. We varied the Poisson's ratio of the reinforcing skeleton by introducing auxetic or honeycomb functional structures that results in Poisson's ratio mismatch between the skeleton and matrix. During uniaxial tensile experiments, high strength and toughness were achieved due to two events: (1) multiple internal bond fractures of the skeleton (like sacrificial bonds in classic DN gels) and (2) significant, biaxial deformation of the matrix imposed by the functional skeleton. The Macro-DN composite with auxetic skeleton exhibits up to 4.2 times higher stiffness and 4.4 times higher yield force than the sum of the component materials. The significant improvement in mechanical performance is correlated to the large mismatch in Poisson's ratio between component materials, and the enhancement is especially noticeable in the low-stretch regime. The strengthening mechanism reported here based on Poisson's ratio mismatch can be widely used for soft materials regardless of chemical composition and will improve the mechanical properties of elastomer and hydrogel systems.

Macroscale Double Networks: Design Criteria for Optimizing Strength and Toughness.

The double network concept, based on the fracture of sacrificial bonds, has been revolutionary toward the creation of robust soft materials. Based on the essence of double network hydrogels, macroscale, three-dimensional printed rigid sacrificial networks are embedded within silicone rubber stretchable matrices. Preferential fracture of the sacrificial network results in a ∼60 time increase in stiffness and a ∼50% increase in the work of extension compared with the neat matrix. Maximizing yield strength while maintaining multistep internal fracture occurs when the strength of the sacrificial network approaches the strength of the matrix. Upon determining the optimal sacrificial network strength, the sacrificial bond section density can be increased to maximize energy dissipation and toughening efficiencies up to ∼70% of the maximum theoretical toughness are achieved. High toughness and dissipation are achieved because topological interlocking enables significant force transmission to the sacrificial network at smaller length scales than interfacial adhesion, allowing much higher sacrificial bond density. This method is general and can be used with a variety of materials systems, without requiring strong interfacial adhesion, contrasting traditional composite systems. Demonstrating that the double network concept can be used at length scales far beyond the molecular scale will have important implications toward the development of future structural materials.

Empirical refinements applicable to the recording of fish sounds in small tanks.

Many underwater bioacoustical recording experiments (e.g., fish sound production during courtship or agonistic encounters) are usually conducted in a controlled laboratory environment of small-sized tanks. The effects of reverberation, resonance, and tank size on the characteristics of sound recorded inside small tanks have never been fully addressed, although these factors are known to influence the recordings. In this work, 5-cycle tone bursts of 1-kHz sound were used as a test signal to investigate the sound recorded in a 170-l rectangular glass tank at various depths and distances from a transducer. The dominant frequency, sound-pressure level, and power spectrum recorded in small tanks were significantly distorted compared to the original tone bursts. Due to resonance, the dominant frequency varied with water depth, and power spectrum level of the projected frequency decreased exponentially with increased distance between the hydrophone and the sound source; however, the resonant component was nearly uniform throughout the tank. Based on the empirical findings and theoretical calculation, a working protocol is presented that minimizes distortion in fish sound recordings in small tanks. To validate this approach, sounds produced by the croaking gourami (Trichopsis vittata) during staged agonistic encounters were recorded according to the proposed protocol in an 1800-l circular tank and in a 37-l rectangular tank to compare differences in acoustic characteristics associated with tank size and recording position. The findings underscore pitfalls associated with recording fish sounds in small tanks. Herein, an empirical solution to correct these distortions is provided.