CRL4 antagonizes SCFFbxo7-mediated turnover of cereblon and BK channel to regulate learning and memory

Intellectual disability (ID), one of the most common human developmental disorders, can be caused by genetic mutations in Cullin 4B (Cul4B) and cereblon (CRBN). CRBN is a substrate receptor for the Cul4A/B-DDB1 ubiquitin ligase (CRL4) and can target voltage- and calcium-activated BK channel for ER retention. Here we report that ID-associated CRL4CRBNmutations abolish the interaction of the BK channel with CRL4, and redirect the BK channel to the SCFFbxo7ubiquitin ligase for proteasomal degradation. Glioma cell lines harbouring CRBN mutations record density-dependent decrease of BK currents, which can be restored by blocking Cullin ubiquitin ligase activity. Importantly, mice with neuron-specific deletion of DDB1 or CRBN express reduced BK protein levels in the brain, and exhibit similar impairment in learning and memory, a deficit that can be partially rescued by activating the BK channel. Our results reveal a competitive targeting of the BK channel by two ubiquitin ligases to achieve exquisite control of its stability, and support changes in neuronal excitability as a common pathogenic mechanism underlying CRL4CRBN–associated ID

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Structural Performance of Composite Shear Walls under Compression

In order to research the effect of different layout forms of steel plate on the axial compression behavior of a steel plate-concrete composite shear wall, this paper presents the experimental results and analysis of the axial compression behavior of a composite shear wall, with different layout forms of steel plate. A total of three tests were carried out, composed of two composite walls with built-in steel plate, and one composite wall with two skins of steel plate. The gross dimensions of the three specimens were 1206 mm × 2006 mm × 300 mm. Experimental results show that the composite wall with two skins of steel plate has an optimal ability of elastic-plastic deformation, and the maximum axial compressive bearing capacity among the three specimens. Using the energy method, the critical local buckling stresses of steel plate were calculated, and compared with the yield stresses. According to different confined actions of concrete, concrete constitutive models were proposed, and the axial compressive strengths of confined concrete were calculated. Considering the local buckling of steel plate and confined concrete, the calculation formula of the axial compression of the composite wall was put forward, and the calculated results were in good agreement with the test results. Therefore, the different layout forms of steel plate have a great influence on its buckling, and on the concrete inhibition effect, which can affect the axial compressive bearing capacity of the composite wall

Oxidative Stress Induced by Metal Ions in Bioleaching of LiCoO2 by an Acidophilic Microbial Consortium

© Copyright © 2020 Liu, Liu, Wu, Zhang, Gu, Zhu and Tan. An acidophilic microbial consortium (AMC) was used to investigate the fundamental mechanism behind the adverse effects of pulp density increase in the bioleaching of waste lithium ion batteries (WLIBs). Results showed that there existed the effect of metal-ion stress on the bio-oxidative activity of AMC. The Li+ and Co2+ accumulated in the leachate were the direct cause for the decrease in lithium and cobalt recovery yields under a high pulp density. In a simulated bioleaching system with 4.0% (w ⋅v–1) LiCoO2, the intracellular reactive oxygen species (ROS) content in AMC increased from 0.82 to 6.02 within 24 h, which was almost three times higher than that of the control (2.04). After the supplementation of 0.30 g⋅L–1 of exogenous glutathione (GSH), the bacterial intracellular ROS content decreased by 40% within 24 h and the activities of intracellular ROS scavenging enzymes, including glutathione peroxidase (GSH-Px) and catalase (CAT), were 1.4- and 2.0-folds higher in comparison with the control within 24 h. In the biofilms formed on pyrite in the bioleaching of WLIBs, it was found that metal-ion stress had a great influence on the 3-D structure and the amount of biomass of the biofilms. After the exogenous addition of GSH, the structure and the amount of biomass of the biofilms were restored to some extent. Eventually, through ROS regulation by the exogenous addition of GSH, very high metal recovery yields of 98.1% Li and 96.3% Co were obtained at 5.0% pulp density

Robust Design Optimization of the Cogging Torque for a PMSM Based on Manufacturing Uncertainties Analysis and Approximate Modeling

A permanent magnet synchronous motor (PMSM) is a crucial device for power conversion in an energy system. The cogging torque of the PMSM is a crucial output characteristic, the robustness of which affects the operational reliability of the energy system. Therefore, the robust design optimization (RDO) of cogging torque has aroused widespread concern. There are several challenges in designing a robust cogging torque PMSM. In particular, some design parameters contain repetitive units, and the finite element analysis (FEA) method is time-consuming. State-of-the-art RDO methods usually treat these uncertainties from repetitive units as the same parameter, which neglects the fluctuation of the manufacturing process and cannot obtain a robust solution for the cogging torque of the motor efficiently and accurately. In order to solve this issue, an approximate modeling method based on manufacturing uncertainties analysis for RDO is proposed in this paper. First, the peak-to-peak value of cogging torque (Tcpp) is used to characterize the cogging torque, which is decoupled to an ideal component and fluctuation component produced by the center values and manufacturing tolerances of design parameters. The design of experiments (DoE) and simulation of the two components are carried out. Then, these two components are approximated separately, and the approximate model of Tcpp is obtained by adding the two components. Finally, the proposed approximate model is embedded into the RDO algorithm, and the PMSM design scheme for good Tcpp robustness is obtained. The effectiveness of the proposed method is verified through a case study of the PMSM

Extracellular Electron Transfer Is a Bottleneck in the Microbiologically Influenced Corrosion of C1018 Carbon Steel by the Biofilm of Sulfate-Reducing Bacterium Desulfovibrio vulgaris.

Carbon steels are widely used in the oil and gas industry from downhole tubing to transport trunk lines. Microbes form biofilms, some of which cause the so-called microbiologically influenced corrosion (MIC) of carbon steels. MIC by sulfate reducing bacteria (SRB) is often a leading cause in MIC failures. Electrogenic SRB sessile cells harvest extracellular electrons from elemental iron oxidation for energy production in their metabolism. A previous study suggested that electron mediators riboflavin and flavin adenine dinucleotide (FAD) both accelerated the MIC of 304 stainless steel by the Desulfovibrio vulgaris biofilm that is a corrosive SRB biofilm. Compared with stainless steels, carbon steels are usually far more prone to SRB attacks because SRB biofilms form much denser biofilms on carbon steel surfaces with a sessile cell density that is two orders of magnitude higher. In this work, C1018 carbon steel coupons were used in tests of MIC by D. vulgaris with and without an electron mediator. Experimental weight loss and pit depth data conclusively confirmed that both riboflavin and FAD were able to accelerate D. vulgaris attack against the carbon steel considerably. It has important implications in MIC failure analysis and MIC mitigation in the oil and gas industry

Integrating serum pharmacochemistry and network pharmacology to reveal the active constituents and mechanism of Corydalis Rhizoma in treating Alzheimer's disease

BackgroundAlzheimer's disease (AD) is a multifactorial neurodegenerative condition. The search for multi-target traditional Chinese medicines or ingredients for treating AD has attracted much attention. Corydalis rhizome (CR) is a traditional Chinese medicine. Its main components are alkaloids, which have therapeutic effects that can potentially be used for treating AD. However, no systematic study has been conducted to explore the anti-AD efficacy of CR, as well as its active compounds and mechanisms of action.ObjectiveThe present study aimed to clarify CR's active constituents and its pharmacological mechanisms in treating AD.MethodsA D-galactose & scopolamine hydrobromide-induced AD mouse model was used and CR was administered orally. The prototypical alkaloid components were identified in the serum. The core components, key targets, and possible mechanisms of action of these alkaloids were revealed through network pharmacology. Molecular docking of the key target was performed. Finally, the mechanism was validated by lipopolysaccharide (LPS)-induced activation of BV2 microglia.ResultsThe results showed that CR improved anxiety-like behavior, spatial and non-spatial recognition, and memory capacity in AD mice. It also achieved synergistic AD treatment by modulating neurotransmitter levels, anti-neuroinflammation, and anti-oxidative stress. The core components that enhance CR's efficacy in treating AD are protoberberine-type alkaloids. The CR may induce the polarization of LPS-activated BV2 microglia from phenotype M1 to M2. This is partially achieved by modulating the IL-6/JAK2/STAT3 signaling pathway, which could be the mechanism by which CR treats AD through anti-inflammation.ConclusionThe present study provided a theoretical and experimental basis for the clinical application of CR in treating AD. It also provides information that aids the secondary development, and precise clinical use of CR

Planktonic <i>D</i>. <i>vulgaris</i> cell counts after 7 days of incubation with and without electron mediators.

<p>Planktonic <i>D</i>. <i>vulgaris</i> cell counts after 7 days of incubation with and without electron mediators.</p

Adaptive bidirectional extracellular electron transfer during accelerated microbiologically influenced corrosion of stainless steel

Microbiologically influenced corrosion is a major source of degradation of metals. Here, extracellular electron transfer is studied during pitting corrosion of stainless steel in the presence of an electroactive bacterium and a riboflavin electron shuttle, revealing bidirectional electron transfer