Pathogenic Connexin-31 Forms Constitutively Active Hemichannels to Promote Necrotic Cell Death

Mutations in Connexin-31 (Cx31) are associated with multiple human diseases including erythrokeratodermia variabilis (EKV). The molecular action of Cx31 pathogenic mutants remains largely elusive. We report here that expression of EKV pathogenic mutant Cx31R42P induces cell death with necrotic characteristics. Inhibition of hemichannel activity by a connexin hemichannel inhibitor or high extracellular calcium suppresses Cx31R42P-induced cell death. Expression of Cx31R42P induces ER stress resulting in reactive oxygen species (ROS) production, in turn, to regulate gating of Cx31R42P hemichannels and Cx31R42P induced cell death. Moreover, Cx31R42P hemichannels play an important role in mediating ATP release from the cell. In contrast, no hemichannel activity was detected with cells expressing wildtype Cx31. Together, the results suggest that Cx31R42P forms constitutively active hemichannels to promote necrotic cell death. The Cx31R42P active hemichannels are likely resulted by an ER stress mediated ROS overproduction. The study identifies a mechanism of EKV pathogenesis induced by a Cx31 mutant and provides a new avenue for potential treatment strategy of the disease

Molecular cloning and characterization of CIDE-3, a novel member of the cell-death-inducing DNA-fragmentation-factor (DFF45)-like effector family.

DNA fragmentation is one of the critical steps in apoptosis, which is induced by DNA fragmentation factor (DFF). DFF is composed of two subunits, a 40 kDa caspase-activated nuclease (DFF40) and a 45 kDa inhibitor (DFF45). Recently a novel family of cell-death-inducing DFF45-like effectors (CIDEs) has been identified. Among CIDEs, two from human (CIDE-A and CIDE-B) and three from mouse (CIDE-A, CIDE-B and FSP27) have been reported. In this study human CIDE-3, a novel member of CIDEs, was identified upon sequence analysis of a previously unidentified cDNA that encoded a protein of 238 amino acids. It was shown to be a human homologue of mouse FSP27, and shared homology with the CIDE-N and CIDE-C domains of CIDEs. Apoptosis-inducing activity was clearly shown by DNA-fragmentation assay of the nuclear DNA of CIDE-3 transfected 293T cells. The expression pattern of CIDE-3 was different from that of CIDE-B. As shown by Northern-blot analysis, CIDE-3 was expressed mainly in human small intestine, heart, colon and stomach, while CIDE-B showed strong expression in liver and small intestine and at a lower level in colon, kidney and spleen. Green-fluorescent-protein-tagged CIDE-3 was revealed in some cytosolic corpuscles. Alternative splicing of the CIDE-3 gene was also identified by reverse transcription PCR, revealing that two transcripts, CIDE-3 and CIDE-3alpha, were present in HepG2 and A375 cells. CIDE-3 comprised a full-length open reading frame with 238 amino acids; in CIDE-3alpha exon 3 was deleted and it encoded a protein of 164 amino acids. Interestingly the CIDE-3alpha isoform still kept the apoptosis-inducing activity and showed the same pattern of subcellular localization as CIDE-3. Consistent with its chromosome localization at 3p25, a region associated with high frequency loss of heterozygosity in many tumours, CIDE-3 may play an important role in prevention of tumorigenesis

Evolution of Microstructures and Mechanical Properties of Nb-V Alloyed Ultra-High Strength Hot Stamping Steel in Austenitizing Process

Clarifying the influence of Nb and V microalloying on the ultra-high strength hot stamping steel (UHSHSS) and exploring appropriate process parameters are the basis for effectively regulating properties of the final product. In this study, the effects of different austenitizing temperatures and holding times on the phase transitions, grain sizes and mechanical properties of 22MnB5NbV with Nb and V alloyed are studied by using JMatPro thermodynamic calculations and experiments. By comparing with 22MnB5 without Nb and V alloyed, the effects of Nb and V elements on quenching microstructures, grain sizes and mechanical properties of UHSHSS are analyzed. The suitable austenitizing process parameters of 22MnB5NbV have been obtained. The results show that the grain size of Nb-V-alloyed UHSHSS grows with the increase in the austenitizing temperature and holding time. The 22MnB5NbV steel can be completely austenitized while the austenitizing temperatures ≥870 °C and holding time ≥3 min. Combined with the actual production process, the best austenitizing temperature and holding time are 930 °C and 3 min. Under these conditions, the average grain size is 7.7 μm, the tensile strength, elongation and strength-ductility product are 1570.8 MPa, 6.6% and 10.4 GPa·%, respectively. With the addition of Nb and V elements, the nanoscale precipitates lead to the refinement of the quenched structure and grain size, and the comprehensive properties of UHSHSS have been effectively promoted, in which the elongation and strong-plastic products are increased by ~0.6% and ~1.2 GPa·%, respectively

ResAttn-recon: Residual self-attention based cortical surface reconstruction

Introduction: The accurate cerebral cortex surface reconstruction is crucial for the study of neurodegenerative diseases. Existing voxelwise segmentation-based approaches like FreeSurfer and FastSurfer are limited by the partial volume effect, meaning that reconstruction details highly rely on the resolution of the input volume. In the computer version area, the signed distance function has become an efficient method for 3D shape representation, the inherent continuous nature makes it easy to capture the fine details of the target object at an arbitrary resolution. Additionally, as one of the most valuable breakthroughs in deep learning research, attention is a powerful mechanism developed to enhance the performance of the encoder-decoder architecture.Methods: To further improve the reconstruction accuracy of the cortical surface, we proposed ResAttn-Recon, a residual self-attention based encoder-decoder framework. In this framework, we also developed a lightweight decoder network with skip connections. Furthermore, a truncated and weighted L1 loss function are proposed to accelerate network convergence, compared to simply applying the L1 loss function.Results: The intersection over union curve in the training process achieved a steeper slope and a higher peak (0.948 vs. 0.920) with a truncated L1 loss. Thus, the average symmetric surface distance (AD) for the inner and outer surfaces is 0.253 ± 0.051 and the average Hausdorff distance (HD) is 0.629 ± 0.186, which is lower than that of DeepCSR, whose absolute distance equals 0.283 ± 0.059 and Hausdorff distance equals 0.746 ± 0.245.Discussion: In conclusion, the proposed residual self-attention-based framework can be a promising approach for improving the cortical surface reconstruction performance

The LIS1/NDE1 Complex Is Essential for FGF Signaling by Regulating FGF Receptor Intracellular Trafficking

Summary: Intracellular transport of membranous organelles and protein complexes to various destinations is fundamental to signaling transduction and cellular function. The cytoplasmic dynein motor and its regulatory proteins LIS1 and NDE1 are required for transporting a variety of cellular cargos along the microtubule network. In this study, we show that deletion of Lis1 in developing lung endoderm and limb mesenchymal cells causes agenesis of the lungs and limbs. In both mutants, there is increased cell death and decreased fibroblast growth factor (FGF) signaling activity. Mechanistically, LIS1 and its interacting protein NDE1/NDEL1 are associated with FGF receptor-containing vesicles and regulate FGF receptor intracellular trafficking and degradation. Notably, FGF signaling promotes NDE1 tyrosine phosphorylation, which leads to dissociation of LIS1/NDE1 complex. Thus, our studies identify the LIS1/NDE1 complex as an important FGF signaling regulator and provide insights into the bidirectional regulation of cell signaling and transport machinery for endocytosis. : Liu et al. show that Lis1 is required for mouse embryonic lung and limb formation. The LIS1/NDE1/dynein complex regulates FGF receptor trafficking and stability. FGF signaling promotes NDE1 tyrosine phosphorylation, which leads to dissociation of the LIS1/NDE1 complex, suggesting a bidirectional regulation of cell signaling and transport machinery. Keywords: LIS1, NDE1, dynein, FGF signaling, development, endocytosi

A 3D-Printed Ferromagnetic Liquid Crystal Elastomer with Programmed Dual-Anisotropy and Multi-Responsiveness

Liquid crystal elastomers (LCE) and magnetic soft materials are promising active materials in many emerging fields, such as soft robotics. Despite the high demand for developing active materials that combine the advantages of LCE and magnetic actuation, the lack of independent programming of the LCE nematic order and magnetization in a single material still hinders the desired multi-responsiveness. In this study, a ferromagnetic LCE (magLCE) ink with nematic order and magnetization is developed that can be independently programmed to be anisotropic, referred to as "dual anisotropy", via a customized 3D-printing platform. The magLCE ink is fabricated by dispersing ferromagnetic microparticles in the LCE matrix, and a 3D-printing platform is created by integrating a magnet with 3-DoF motion into an extrusion-based 3D printer. In addition to magnetic fields, magLCEs can also be actuated by heating sources (either environmental heating or photo-heating of the embedded ferromagnetic microparticles) with a high energy density and tunable actuation temperature. A programmed magLCE strip robot is demonstrated with enhanced adaptability to complex environments (different terrains, magnetic fields, and temperatures) using a multi-actuation strategy. The magLCE also has potential applications in mechanical memory, as demonstrated by the multistable mechanical metastructure array with remote writability and stable memory. A ferromagnetic liquid crystal elastomer (magLCE) ink whose nematic order and magnetization can be independently programmed to be anisotropic via a customized 3D-printing platform is reported. The 3D-printed magLCE provides designable multimodal shape morphing under different stimuli, which is promising for wireless soft robots and intelligent devices.imag

Identification and analysis of key hypoxia- and immune-related genes in hypertrophic cardiomyopathy

Abstract Background Hypertrophic cardiomyopathy (HCM), an autosomal dominant genetic disease, is the main cause of sudden death in adolescents and athletes globally. Hypoxia and immune factors have been revealed to be related to the pathology of HCM. There is growing evidence of a role for hypoxia and inflammation as triggers and enhancers in the pathology in HCM. However, the role of hypoxia- and immune-related genes in HCM have not been reported. Methods Firstly, we obtained four HCM-related datasets from the Gene Expression Omnibus (GEO) database for differential expression analysis. Immune cells significantly expressed in normal samples and HCM were then screened by a microenvironmental cell population counter (MCP-counter) algorithm. Next, hypoxia- and immune-related genes were screened by the LASSO + support vector machine recursive feature elimination (SVM-RFE) and weighted gene co-expression network analysis (WGCNA). Single-gene enrichment analysis and expression validation of key genes were then performed. Finally, we constructed a competing endogenous RNA (ceRNA) network of key genes. Results In this study, 35 differentially expressed hypoxia genes were found. By using LASSO + SVM-RFE analysis, 10 more targets with differentially expressed hypoxia genes were identified. The MCP-count algorithm yielded five differentially expressed immune cells, and after assessing them for WGCNA characteristics, 612 immune genes were discovered. When hypoxia and immune genes were combined for cross-tabulation analysis, three hypoxia- and immune-related genes (ATP2A2, DDAH1, and OMA1) were identified. Conclusion Based on hypoxia characteristic genes, three key genes were identified. These were also significantly related to immune activation, which proves a theoretical basis and reference value for studying the relationship between HCM and hypoxia and immunity. Graphical Abstrac