Focal calcium monitoring with targeted nanosensors at the cytosolic side of endoplasmic reticulum

Ca²⁺ distribution is spatially and temporally non-uniform inside cells due to cellular compartmentalization. However, Ca²⁺ sensing with small organic dyes, such as fura-2 and fluo-4, has been practically applied at a single cell level where the averaged signal from freely diffusing dye molecules is acquired. In this study, we aimed to target azide-functionalized fura-2 (N₃-fura-2) to a specific site of subcellular compartments to realize focal Ca²⁺ sensing. Using scAVD (single-chain avidin)–biotin interaction and a copper-free click reaction system, we linked N₃-fura-2 to specifically-targeted scAVD protein fused with a red fluorescent protein mCherry, so that Ca²⁺ sensors conjugated with four N₃-fura-2 dyes with dibenzocyclooctyne (DBCO)-PEG4-biotin as a linker were generated at subcellular compartments in living cells. In cytoplasm, N₃-fura-2 showed a prolonged retention period after binding to scAVD. Furthermore, the reacted N₃-fura-2 was retained inside cells even after free dyes were washed out by methanol fixation. When scAVD was overexpressed on endoplasmic reticulum (ER) membranes, N₃-fura-2 was accumulated on ER membranes. Upon histamine stimulation, which increases cytosolic Ca²⁺ concentration, ER-localized N₃-fura-2 successfully sensed the Ca²⁺ level changes at the cytosolic side of ER membrane. Our study demonstrated specific targeting of N₃-fura-2 to subcellular compartments and the ability of sensing focal Ca²⁺ level changes with the specifically targeted Ca²⁺ sensors.     

Acceleration of the Rate of Silver Nanoparticle Formation Using Microbubbles in a Sonochemical Process

The effect of the addition of microbubbles on the formation of silver nanoparticles produced in an ultrasonic radiation-mediated process was investigated. The addition of microbubbles with an area-based median diameter of 26.0?µm and a bubble volume density of 0.18?cm³/L significantly increased the silver nanoparticle formation rate during the sonochemical process. The size distribution of the silver nanoparticles was largely unaffected by the addition of the microbubbles. The influence of changes in the microbubble volume density on the formation of the silver nanoparticles was also investigated; it was confirmed that the rate of formation of the silver nanoparticles increased moderately with increasing volume density. The gradient in absorption spectra was approximately 30 times larger for the case when microbubbles (median diameter: 28.3?µm) were added at 0.74?cm³/L, compared with the case without microbubbles. The results suggested that the microbubbles provided reaction sites similar to cavitation bubbles in the ultrasonic reaction.     

Analysis and modeling of char particle combustion with heat and multicomponent mass transfer

A char combustion model suitable for a large-scale boiler/gasifier simulation, which considers the variation of physical quantities in the radial direction of char particles, is developed and examined. The structural evolution within particles is formulated using the basic concept of the random pore model while simultaneously considering particle shrinkage. To reduce the computational cost, a new approximate analytical boundary condition is applied to the particle surface, which is approximately derived from the Stefan–Maxwell equations. The boundary condition showed reasonably good agreement with direct numerical integration with a fine grid resolution by the finite difference method under arbitrary conditions. The model was applied to combustion in a drop tube furnace and showed qualitatively good agreement with experiments, including for the burnout behavior in the late stages. It is revealed that the profiles of the oxygen mole fraction, conversion, and combustion rate have considerably different characteristics in small and large particles. This means that a model that considers one total conversion for each particle is insufficient to describe the state of particles. Since our char combustion model requires only one fitting parameter, which is determined from information on the internal geometry of char particles, it is useful for performing numerical simulations.     

Aerobic Exercise Ameliorates Cancer Cachexia-Induced Muscle Wasting through Adiponectin Signaling

Cachexia is a multifactorial syndrome characterized by muscle loss that cannot be reversed by conventional nutritional support. To uncover the molecular basis underlying the onset of cancer cachectic muscle wasting and establish an effective intervention against muscle loss, we used a cancer cachectic mouse model and examined the effects of aerobic exercise. Aerobic exercise successfully suppressed muscle atrophy and activated adiponectin signaling. Next, a cellular model for cancer cachectic muscle atrophy using C2C12 myotubes was prepared by treating myotubes with a conditioned medium from a culture of colon-26 cancer cells. Treatment of the atrophic myotubes with recombinant adiponectin was protective against the thinning of cells through the increased production of p-mTOR and suppression of LC3-II. Altogether, these findings suggest that the activation of adiponectin signaling could be part of the molecular mechanisms by which aerobic exercise ameliorates cancer cachexia-induced muscle wasting.     

Microencapsulation of bioactive compounds from mulberry (Morus alba L.) leaf extracts by protein–polysaccharide interactions

Mulberry leaf extracts were generated using four concentrations of ethanol (50%, 60%, 70%, and 95% v/v). A 60% ethanolic mulberry leaf extract (60E) yielded a high total phenolic content (TPC) and antioxidant activity using 1, 1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging and a ferric reducing antioxidant power (FRAP) assay. Coating materials were derived using a combination of soy protein isolates (SPI) and low methoxyl (LM) pectin in a 1:1 ratio. The effect of various parameters on microencapsulation, such as pH (3.5, 4.0, and 4.5) and the concentration of coating materials (2.5, 5.0 and 7.5% w/v), was studied. Microcapsules produced using 60E as a core material at pH 4.0 with 7.5% of coating material showed a high encapsulation yield, encapsulation efficiency, TPC and antioxidant activity.     

The Critical Energy Release Rate of Welded Joints Between Fiber-Reinforced Thermoplastics and Metals When Thermal Residual Stress is Considered

Thermal effects on welded joints between fiber-reinforced thermoplastics (FRTPs) and metals have been investigated theoretically and experimentally. Because FRTPs use thermoplastics as the matrix resin, they have advantages over fiber-reinforced thermoset plastics (FRPs or FRSPs), including the ability to be welded. When dissimilar materials are welded together, however, thermal stress occurs due to the different thermal expansions of the materials and affects the energy release rate of the joint. Therefore, a method for evaluating the true energy release rate, including the effect of thermal stress, is necessary for strength evaluation tests. Although several theories that compensate for the thermal stress and evaluate the true energy release rate have already been proposed, they require parameters that are difficult to measure. Therefore, it is difficult to apply them in experimental investigations. In this article, a theoretical method with easily measurable parameters is proposed to calculate the energy release rate of welded double cantilever beam (DCB) joints. The effect of the thermal stress on the critical energy release rate is discussed in terms of the experimental results of a welded DCB specimen composed of a FRTP and an aluminum alloy.     

Stabilizing effect of the cyclodextrins additive to spray-dried particles of curcumin/polyvinylpyrrolidone on the supersaturated state of curcumin

Although curcumin is considered to have various therapeutic effects, its use as a functional food or supplement is restricted owing to its low water solubility and bioavailability. To increase the solubility of curcumin in water, the use of polyvinylpyrrolidone (PVP) and vinylpyrrolidone-vinyl acetate copolymers with a pyrrolidone skeleton was noted to be promising. In particular, the bi-component formulations of curcumin/PVP prepared through spray drying exhibited an amorphous state in powder X-ray diffraction observations and temporally increased the apparent solubility of curcumin to over 5000 times that of untreated curcumin; nevertheless, after 24 h, the solubility decreased owing to the unstable supersaturated state of curcumin. The addition of α-cyclodextrin (α-CyD) in the bi-component curcumin/PVP formulation helped maintain the supersaturated state of curcumin, whereas the addition of β- and γ-CyD led to the collapse of the supersaturated state. The addition of α-CyD can likely help inhibit the nucleation and crystal growth of curcumin, through the interaction among the solubilized units of curcumin/PVP and α-CyD.