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Calmodulin transduces [Ca2+] information regulating the rhythmic Ca2+ cycling between the sarcoplasmic reticulum and cytoplasm during contraction and relaxation in cardiac and skeletal muscle. However, the structural dynamics by which calmodulin modulates the sarcoplasmic reticulum Ca2+ release channel, the ryanodine receptor, at physiologically relevant [Ca2+] is unknown. Using fluorescence lifetime FRET, we resolve different structural states of calmodulin and Ca2+-driven shifts in the conformation of calmodulin bound to ryanodine receptor. Skeletal and cardiac ryanodine receptor isoforms show different calmodulin-ryanodine receptor conformations, as well as binding and structural kinetics with 0.2-ms resolution, which reflect different functional roles of calmodulin. These FRET methods provide insight into the physiological calmodulin-ryanodine receptor structural states, revealing additional distinct structural states that complement cryo-EM models that are based on less physiological conditions. This technology will drive future studies on pathological calmodulin-ryanodine receptor interactions and dynamics with other important ryanodine receptor bound modulators.

The N-terminal region (NTR) of ryanodine receptor (RyR) channels is critical for the regulation of Ca2+ release during excitation-contraction (EC) coupling in muscle. The NTR hosts numerous mutations linked to skeletal (RyR1) and cardiac (RyR2) myopathies, highlighting its potential as a therapeutic target. Here, we constructed two biosensors by labeling the mouse RyR2 NTR at domains A, B, and C with FRET pairs. Using fluorescence lifetime (FLT) detection of intramolecular FRET signal, we developed high-throughput screening (HTS) assays with these biosensors to identify small-molecule RyR modulators. We then screened a small validation library and identified several hits. Hits with saturable FRET dose-response profiles and previously unreported effects on RyR were further tested using [3H]ryanodine binding to isolated sarcoplasmic reticulum vesicles to determine effects on intact RyR opening in its natural membrane. We identified three novel inhibitors of both RyR1 and RyR2 and two RyR1-selective inhibitors effective at nanomolar Ca2+. Two of these hits activated RyR1 only at micromolar Ca2+, highlighting them as potential enhancers of excitation-contraction coupling. To determine whether such hits can inhibit RyR leak in muscle, we further focused on one, an FDA-approved natural antibiotic, fusidic acid (FA). In skinned skeletal myofibers and permeabilized cardiomyocytes, FA inhibited RyR leak with no detrimental effect on skeletal myofiber excitation-contraction coupling. However, in intact cardiomyocytes, FA induced arrhythmogenic Ca2+ transients, a cautionary observation for a compound with an otherwise solid safety record. These results indicate that HTS campaigns using the NTR biosensor can identify compounds with therapeutic potential.

The preparation of flawless and defect-free mixed matrix membranes (MMMs) comprising metal-organic framework (MOF) and polymer is often difficult owing to the poor MOF/polymer interface compatibility. Herein, we present the synthesis of an important family of pillared-layered MOFs with polymerizable moieties based on the parent structure [Zn2L2P]n [L = vinyl containing benzenedicarboxylic acid linkers; P = 4,4'-bipyridine (bipy)]. The crystalline structures of polymerizable MOFs were analyzed using single-crystal X-ray crystallography. The presence of reactive double bonds in MOFs was verified by the successful thiol-ene click reaction with sulfhydryl compounds. The subsequent copolymerization of polymerizable MOFs with organic monomers produced mixed matrix membranes with enhanced MOF/polymer interfacial adhesion that enabled good separation efficiency of CO2 from flue gas. This strategy provides a stimulating platform to the preparation of highly efficient MMMs that are capable of mitigating energy consumption and environment issues.

In EV/HEV application, the maximum internal temperature allowed for film capacitor should be less than 105 ℃, which leaves especial limited temperature margin as the capacitor works in high ambient temperature of 85 ℃. However, temperature rising of capacitor is strongly related to the load condition. And it is difficult to define the temperature allowance of capacitor accurately in the early design. For this purpose, a consolidated electrothermal simulation method based Foster thermal network model is presented in this paper to predict the transient temperature inside the capacitor. To set up a complete simulation model, the parameters of five-order Foster resistor-capacitor thermal network model are extracted by the method of capacitor temperature rise experiment. The temperature rising of film capacitor in the conditions of NEDC (New European Driving Cycle) & USA highway driving cycle is calculated by the complete simulation model. Simulation results show that a good correspondence is presented between capacitor temperature rising curve vs ripple current under driving cycle, and the maximum temperature rise can be clearly assessed by this temperature prediction method.

In the field of cement, if the formation cannot be given sufficient pressure to maintain stability during construction, pressure control failure may occur, leading to the leakage of liquids and gasses from the formation to the wellbore. In addition, irregular wellbore diameter and casing eccentricity are important factors that are easily overlooked and affect the prediction of ECD (Equivalent Circulation Density) calculation. This results in major accidents and ecological disasters, further impacting the global environment. This study focuses on a well in the eastern oilfields of China, and based on a rheological experiment of high temperature and high pressure, an irregular eccentric wellbore model is established according to the measured wellbore diameter and eccentricity data to calculate the ECD of the whole cementing process. Then, a data set is constructed and analyzed using the random forest method to quantitatively evaluate influencing factors such as displacement, rheology, density, and eccentricity on the bottomhole and wellbore ECD. Results find that the density of cement slurry and drilling fluid has the most significant impact on the maximum ECD, with the impact reaching 0.3142 and 0.2902, respectively, and the main factors that affect the minimum ECD are the density and rheological changes in the drilling fluid, reaching 0.7014 and 0.2846. These research findings will contribute to the precise control of wellbore pressure during cementing operations, further ensuring the safety of cementing operations, and laying a technical foundation for the automation and intelligentization of subsequent cementing operations. [ABSTRACT FROM AUTHOR]

Cordyceps militaris solid medium polysaccharides (CMMPs) were extracted using an ultrasound-assisted enzyme method, and the process conditions were optimized via response surface methodology (RSM). The CMMPs were separated into four components named CMMP−1, CMMP−2, CMMP−3 and CMMP−4 using ethanol fractional precipitation, and their monosaccharide composition and structural properties were analyzed by molecular weight analysis, Fourier-transform infrared spectroscopy (FT–IR), scanning electron microscopy (SEM), Congo red test, ultraviolet–visible spectroscopy (UV-vis), atomic force microscopy (AFM), and thermogravimetric analysis (TGA). RSM could predict the yield of the CMMP (R2 = 0.9928), and the polysaccharide yield was 15.43% under the selected conditions of 3.1% cellulase enzyme addition, a liquid–solid ratio of 42:1, an extraction temperature of 61 °C, and an extraction time of 60 min. Glucose and galactose were the main constituents of the four fractional precipitated polysaccharides. Furthermore, four components exhibited antioxidant activity, and CMMP−1 demonstrated stronger antioxidant activity in vitro. This study demonstrates the possibility of developing a natural antioxidant food from Cordyceps militaris solid medium. [ABSTRACT FROM AUTHOR]

This article was designed to explore the effects and mechanisms of miR-185-5p on the replication of hepatitis C virus (HCV). Quantitative reverse transcription PCR (qRT-PCR) was performed for detecting the abundance of miR-185-5p and HCV RNA in HCV-infected primary hepatocytes and Huh7.5 cells. Dual-luciferase reporter gene assay was used for exploring the interaction between miR-185-5p and GALNT8. Western blot analyzed protein expression of GALNT8, NS3, and NS5A. miR-185-5p was remarkably downregulated in HCV-infected primary hepatocytes and Huh7.5 cells. miR-185-5p upregulation inhibited HCV RNA expression, while its inhibition promoted HCV replication. miR-185-5p induced accumulation of NS3 and NS5A in the cells. Dual-luciferase reporter gene assay verified the targeted relationship between miR-185-5p and GALNT8. In addition, the effects of overexpressing or knocking down miR-185-5p on HCV replication could be correspondingly eliminated by the overexpression or knockdown of GALNT8. miR-185-5p may target GALNT8 in JFH1-infected Huh7.5 cells and then inhibit HCV replication. miR-185-5p may be a potential target for treating HCV.

A tremendous amount of research has witnessed the exploration of two-dimensional (2D) materials with intrinsic ferromagnetism and diverse physical properties. However, the low Curie temperature and deficient magnetic anisotropy hinder their practical applications in nanoscale spintronics. Based on first-principles calculations, we propose a new family of 2D ternary transition metal tellurides, CrX2Te4 (X = Al, Ga, and In), with both structural and magnetic stabilities at room temperature. Our calculations demonstrate that the 2D CrX2Te4 crystal exhibits the intrinsic 100% spin-polarized half-metallic feature with spin-up metallic and spin-down semi-conducting properties. With the remarkable magnetic moment of 4 μB per Cr atom, both 2D CrAl2Te4 and CrGa2Te4 crystals perform robust ferromagnetism with the out-of-plane magnetic anisotropy, while the 2D CrIn2Te4 crystal prefers the in-plane easy magnetization axis. The Monte Carlo simulation based on the 2D Heisenberg model shows that the critical Curie temperatures of the 2D CrAl2Te4, CrGa2Te4, and CrIn2Te4 crystals could reach 466, 431, and 536 K, respectively. Moreover, the magnetic exchange strength and magnetic anisotropy could be further enhanced by the in-plane biaxial strain. The novel electronic and magnetic features promote 2D CrX2Te4 (X = Al, Ga, and In) crystals as a new family of two-dimensional intrinsic ferromagnetic materials for next-generation advanced spintronics. [ABSTRACT FROM AUTHOR]

An increasing amount of evidence indicates that Baicalin (Bai, a natural glycosyloxyflavone compound) exhibits an antiviral effect against avian viruses. However, it remains unclear if the antiviral effect of Bai against infectious bronchitis virus (IBV) is exerted indirectly by modulating respiratory tract microbiota and/or their metabolites. In this study, we investigated the protection efficacy of Bai in protecting cell cultures and broilers from IBV infection and assessed modulation of respiratory tract microbiota and metabolites during infection. Bai was administered orally to broilers by being mixed in with drinking water for seven days. Ultimately, broilers were challenged with live IBV. The results showed that Bai treatment reduced respiratory tract symptoms, improved weight gain, slowed histopathological damage, reduced virus loads and decreased pro-inflammation cytokines production. Western blot analysis demonstrated that Bai treatment significantly inhibited Toll-like receptor 7 (TLR7), myeloid differentiation factor 88 (MyD88) and nuclear factor kappa-B (NF-κB) expression both in cell culture and cells of the trachea. Bai treatment reversed respiratory tract microbiota dysbiosis, as shown by 16S rDNA sequencing in the group of broilers inoculated with IBV. Indeed, we observed a decrease in Proteobacteria abundance and an increase in Firmicutes abundance. Metabolomics results suggest that the pentose phosphate pathway, amino acid and nicotinamide metabolism are linked to the protection conferred by Bai against IBV infection. In conclusion, these results indicated that further assessment of anti-IBV strategies based on Bai would likely result in the development of antiviral molecule(s) which can be administered by being mixed with feed or water. [ABSTRACT FROM AUTHOR]

This document is an editorial for a special issue of the journal Micromachines on flexible sensors and actuators for biomedicine. Flexible sensors and actuators are used in wearable devices, implantable medical devices, and micro-robotics for biomedical monitoring, diagnosis, and therapies. The special issue includes research and reviews on biochemical sensors, self-powered sensors driven by nanogenerators, and soft robotics for colonoscopes. The biochemical sensors detect and measure specific chemical substances or biomolecules and have applications in various fields, including biomedicine. The self-powered sensors use nanogenerators to convert mechanical energy into electricity, providing an alternative power supply for micro-devices or sensors. Soft actuators are capable of producing motion or deformation under various stimuli and are used in biomedical applications such as drug delivery and vascular occlusion therapy. The special issue explores the working mechanisms, application scenarios, and potential challenges of these flexible sensors and actuators. [Extracted from the article]