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Although the mechanism for activation of latent TGFβ1 and TGFβ3 is understood to involve the binding of the TGFβ propeptide (LAP) to both an integrin and an insoluble substrate, the activation of latent TGFβ2 has been unclear because the TGFβ2 LAP does not have the classical integrin binding sequence found in the other two TGFβ isoform LAPs. To assess the potential requirement for covalent linkage with a matrix or cell surface protein for the activation of latent TGFβ2, we generated mice in which the TGFβ2 Cys residue predicted to be involved in binding was mutated to Ser (Tgfb2C24S). We reasoned that, if covalent interaction with a second molecule is required for latent TGFβ2 activation, mutant mice should display a Tgfb2 null (Tgfb2−/−)-like phenotype. Tgfb2C24S mice closely phenocopy Tgfb2−/− mice with death in utero between E18 and P1 and with congenital heart and kidney defects similar to those described for Tgfb2−/− mice. The mutant latent TGFβ2 is secreted at levels similar to WT, yet TGFβ signaling monitored as nuclear pSmad2 is suppressed. We conclude that, like latent TGFβ1, latent TGFβ2 activation requires binding to an immobilized matrix or plasma membrane molecule.

Abstract Inter-organelle contact and communication between mitochondria and sarco/endoplasmic reticulum (SR/ER) maintain cellular homeostasis and are profoundly disturbed during tissue ischemia. We tested the hypothesis that the formin Diaphanous-1 (DIAPH1), which regulates actin dynamics, signal transduction and metabolic functions, contributes to these processes. We demonstrate that DIAPH1 interacts directly with Mitofusin-2 (MFN2) to shorten mitochondria-SR/ER distance, thereby enhancing mitochondria-ER contact in cells including cardiomyocytes, endothelial cells and macrophages. Solution structure studies affirm the interaction between the Diaphanous Inhibitory Domain and the cytosolic GTPase domain of MFN2. In male rodent and human cardiomyocytes, DIAPH1-MFN2 interaction regulates mitochondrial turnover, mitophagy, and oxidative stress. Introduction of synthetic linker construct, which shorten the mitochondria-SR/ER distance, mitigated the molecular and functional benefits of DIAPH1 silencing in ischemia. This work establishes fundamental roles for DIAPH1-MFN2 interaction in the regulation of mitochondria-SR/ER contact networks. We propose that targeting pathways that regulate DIAPH1-MFN2 interactions may facilitate recovery from tissue ischemia.

Abstract Synergistically improving T-cell responsiveness is promising for favorable therapeutic outcomes in immunologically cold tumors, yet current treatments often fail to induce a cascade of cancer-immunity cycle for effective antitumor immunity. Gasdermin-mediated pyroptosis is a newly discovered mechanism in cancer immunotherapy; however, cleavage in the N terminus is required to activate pyroptosis. Here, we report a single-agent mRNA nanomedicine-based strategy that utilizes mRNA lipid nanoparticles (LNPs) encoding only the N-terminus of gasdermin to trigger pyroptosis, eliciting robust antitumor immunity. In multiple female mouse models, we show that pyroptosis-triggering mRNA/LNPs turn cold tumors into hot ones and create a positive feedback loop to promote antitumor immunity. Additionally, mRNA/LNP-induced pyroptosis sensitizes tumors to anti-PD-1 immunotherapy, facilitating tumor growth inhibition. Antitumor activity extends beyond the treated lesions and suppresses the growth of distant tumors. We implement a strategy for inducing potent antitumor immunity, enhancing immunotherapy responses in immunologically cold tumors.

Abstract Combined with the spatial data processing capability of the Geographic Information Systems (GIS), the Pan Jiazheng method is extended from two-dimensional (2D) to three-dimensional (3D), and a 3D landslide surge height calculation method is proposed based on grid column units. First, the data related to the landslide are rasterized to form grid columns, and a force analysis model of 3D landslides is established. Combining the vertical strip method with Newton's laws of motion, dynamic equilibrium equations are established to solve the surge height. Moreover, a 3D landslide surge height calculation expansion module is developed in the GIS environment, and the results are compared with those of the 2D Pan Jiazheng method. Comparisons showed that the maximum surge height obtained by the proposed method is 24.6% larger than that based on the Pan Jiazheng method. Compared with the traditional 2D method, the 3D method proposed in this paper better represent the actual spatial state of the landslide and is more suitable for risk assessment.

For the limit equilibrium method based on the assumption model of sliding surface normal stress, the more reasonable the assumed sliding surface normal stress model is, the higher the accuracy of the calculation results will be, which is of great significance to improve the theoretical calculation accuracy. Combined with the powerful spatial data analysis ability of Geographic Information Systems (GIS), the expression of the normal stress distribution function of the slip surface is deduced by establishing a GIS-based three-dimensional slope stability analysis model and limit equilibrium equations. By analyzing the composition of the normal stress distribution on the sliding surface, the composition of the normal stress on the sliding surface is obtained, and then the assumed model of the normal stress distribution on the sliding surface is constructed. Finally, the assumed GIS model of the sliding surface normal stress distribution is verified by calculating the proportion. The model overcomes the problem that the stability factor of the slope will cause large errors in the three-dimensional state, and provides a theoretical calculation basis for the establishment of a three-dimensional slope limit equilibrium method based on the assumption of normal stress on the slip surface in GIS.

To overcome the detrimental effect of residual stress in welded joints, which affects the overall performance of the welded structure, this paper studies the magnitude and distribution of residual stress after welding and local post-weld heat treatment (PWHT). The coupled thermo-metallurgical-mechanical model for welding 6 mm thick Ti-6Al-4V (TC4) titanium alloy plates was established, the evolution of the SSPT and its effect on the residual stress were quantitatively analyzed, and a parametric analysis of local PWHT was performed. The results demonstrated that there was good agreement between the numerical results and the experimental data. Due to the cooling rate reaching 327 °C/s, the volume fraction of α、 in the fusion zone (FZ) reached 0.218 after welding and decreased by 90.83% after PWHT when the heating temperature was 700 °C. The peak value of the longitudinal residual stress can reach 686.4 MPa after welding with SSPT, which was 11.38% lower than that without SSPT, and it decreased by 65.6% after PWHT when the heating temperature was 900 °C. The research results demonstrate that SSPT has a significant effect on residual stress, and PWHT can obviously reduce the residual stress, which provides a certain reference for welding TC4 titanium alloy plates.

Research on the deformation and failure mechanism of rock slopes in high-altitude cold areas has obtained certain results, but based on the current theory and technology, it is difficult to comprehensively solve problems related to the cold-slope instability mechanisms and disaster prevention and control. As yet, no overall research system for high-altitude slope mining or criteria for slope stability have been established. In this paper, based on an extensive literature review, five measures of the deformation and failure of alpine rock slopes were presented, including the indoor rock mechanics test, simulation of physically similar slopes, multi-field multi-phase coupled numerical simulation, in-situ monitoring of deformation and damage, and the instability mechanism of rock slopes in high-altitude cold areas. After summarizing the research results related to the deformation and failure of alpine rock masses, existing problems were discussed and current research deficiencies were analyzed. The key problems that require urgent solutions in the research of the deformation and failure of alpine rock slopes were summarized. The first problem is the damage mechanism of a rock mass in a high-altitude cold area under mining disturbance conditions. The second problem is the aging characteristics and evaluation methods of the instability of a flow–solid–gas multi-phase multi-field coupled slope under freeze–thaw cycles. The future research direction and development trends in the deformation and failure of rock slopes in cold regions with high altitude were also analyzed. Research should be conducted on the following: (1) the damage degradation mechanism of rock masses with different stress paths coupled with freeze–thaw cycles, (2) the structural plane collapse mechanism and instability of rock slopes in high-altitude cold areas under the condition of blast mining, (3) the dynamic response and disaster occurrence law of jointed rock slopes in high-altitude cold areas under earthquake loading, (4) the mechanism of damage deterioration of jointed rock masses under multi-field and multi-phase coupling conditions, and (5) real-time safety monitoring and early-warning technology regarding the instability of multi-parameter cold resistance of mine slopes in high-altitude cold areas. These five research areas constitute the trends of future research.

The IFTTT style event-driven programming paradigm benefits normal users to build their own customized home automation Internet of things (IoT) system,meanwhile,it also brings serious safety and security risks.To handle this problem,Menshen was designed and implemented,an automated configuration,simulation and verification platform for event-driven home automation IoT system based on model checking.Users can easily set up their own smart home systems in Menshen,and conduct simulation and verification in a push-button style.Menshen could further demonstrate the error trace to help users to understand the behavior of the system and increase the safety and security of the system.An experiment with a large number of cases is carried out,and the results show that 86.7% cases are error-prone,and the verification only took 0.7 seconds in average.

Background/Aims: Autosomal dominant polycystic kidney disease (ADPKD) is the most common genetic form of kidney disease. High-throughput microarray analysis has been applied for elucidating key genes and pathways associated with ADPKD. Most genetic profiling data from ADPKD patients have been uploaded to public databases but not thoroughly analyzed. This study integrated 2 human microarray profile datasets to elucidate the potential pathways and protein-protein interactions (PPIs) involved in ADPKD via bioinformatics analysis in order to identify possible therapeutic targets. Methods: The kidney tissue microarray data of ADPKD patients and normal individuals were searched and obtained from NCBI Gene Expression Omnibus. Differentially expressed genes (DEGs) were identified, and enriched pathways and central node genes were elucidated using related websites and software according to bioinformatics analysis protocols. Seven DEGs were validated between polycystic kidney disease and control kidney samples by quantitative real-time polymerase chain reaction. Results: Two original human microarray datasets, GSE7869 and GSE35831, were integrated and thoroughly analyzed. In total, 6,422 and 1,152 DEGs were extracted from GSE7869 and GSE35831, respectively, and of these, 561 DEGs were consistent between the databases (291 upregulated genes and 270 downregulated genes). From 421 nodes, 34 central node genes were obtained from a PPI network complex of DEGs. Two significant modules were selected from the PPI network complex by using Cytotype MCODE. Most of the identified genes are involved in protein binding, extracellular region or space, platelet degranulation, mitochondrion, and metabolic pathways. Conclusions: The DEGs and related enriched pathways in ADPKD identified through this integrated bioinformatics analysis provide insights into the molecular mechanisms of ADPKD and potential therapeutic strategies. Specifically, abnormal decorin expression in different stages of ADPKD may represent a new therapeutic target in ADPKD, and regulation of metabolism and mitochondrial function in ADPKD may become a focus of future research.

Monogenic disorders (MGDs), which are caused by single gene mutations, have a serious effect on human health. Among these, β-thalassemia (β-thal) represents one of the most common hereditary hematological diseases caused by mutations in the human hemoglobin β (HBB) gene. The technologies of induced pluripotent stem cells (iPSCs) and genetic correction provide insights into the treatments for MGDs, including β-thal. However, traditional approaches for correcting mutations have a low efficiency and leave a residual footprint, which leads to some safety concerns in clinical applications. As a proof of concept, we utilized single-strand oligodeoxynucleotides (ssODNs), high-fidelity CRISPR/Cas9 nuclease, and small molecules to achieve a seamless correction of the β-41/42 (TCTT) deletion mutation in β thalassemia patient-specific iPSCs with remarkable efficiency. Additionally, off-target analysis and whole-exome sequencing results revealed that corrected cells exhibited a minimal mutational load and no off-target mutagenesis. When differentiated into hematopoietic progenitor cells (HPCs) and then further to erythroblasts, the genetically corrected cells expressed normal β-globin transcripts. Our studies provide the most efficient and safe approach for the genetic correction of the β-41/42 (TCTT) deletion in iPSCs for further potential cell therapy of β-thal, which represents a potential therapeutic avenue for the gene correction of MGD-associated mutants in patient-specific iPSCs. Keywords: β thalassemia, induced pluripotent stem cells, CRISPR/Cas9, ssODNs

Understanding coronary vessels development provides basis for regenerative strategies. Here, Soh et al. identify endothelin-1 as a key molecule driving long-term expansion of ISL1+bipotent vascular progenitors derived from human embryonic stem cells, and show that these cells can regenerate coronary vessels in mice.

With recent advances in stem cell technology, it is becoming efficient to differentiate human pluripotent stem cells (hPSCs) into cardiomyocytes, which can subsequently be used for myriad purposes, ranging from interrogating mechanisms of cardiovascular disease, developing novel cellular therapeutic approaches, as well as assessing the cardiac safety profile of compounds. However, the relative inability to acquire abundant pure and mature cardiomyocytes still hinders these applications. Recently, it was reported that glucose-depleted culture medium supplemented with lactate can facilitate purification of hPSC-derived cardiomyocytes. Here, we report that fatty acid as a lactate replacement has not only a similar purification effect but also improves the electrophysiological characteristics of hPSC-derived cardiomyocytes. Glucose-depleted culture medium supplemented with fatty acid and 3,3′,5-Triiodo-l-thyronine (T3) was used during enrichment of hPSC-derived cardiomyocytes. Compared to untreated control cells, the treated cardiomyocytes exhibited enhanced action potential (AP) maximum upstroke velocity (as shown by a significant increase in dV/dtmax), action potential amplitude, as well as AP duration at 50% (APD50) and 90% (APD90) of repolarization. The treated cardiomyocytes displayed higher sensitivity to isoproterenol, more organized sarcomeric structures, and lower proliferative activity. Expression profiling showed that various ion channel and cardiac-specific genes were elevated as well. Our results suggest that the use of fatty acid and T3 can facilitate purification and maturation of hPSC-derived cardiomyocytes.

Diabetic cardiomyopathy is a complication of type 2 diabetes, with known contributions of lifestyle and genetics. We develop environmentally and genetically driven in vitro models of the condition using human-induced-pluripotent-stem-cell-derived cardiomyocytes. First, we mimic diabetic clinical chemistry to induce a phenotypic surrogate of diabetic cardiomyopathy, observing structural and functional disarray. Next, we consider genetic effects by deriving cardiomyocytes from two diabetic patients with variable disease progression. The cardiomyopathic phenotype is recapitulated in the patient-specific cells basally, with a severity dependent on their original clinical status. These models are incorporated into successive levels of a screening platform, identifying drugs that preserve cardiomyocyte phenotype in vitro during diabetic stress. In this work, we present a patient-specific induced pluripotent stem cell (iPSC) model of a complex metabolic condition, showing the power of this technique for discovery and testing of therapeutic strategies for a disease with ever-increasing clinical significance.

Compared with traditional conductive fillers, carbon nanotubes (CNTs) have unique advantages, i.e., excellent mechanical properties, high electrical conductivity and thermal stability. Nanocomposites as piezoresistive films provide an interesting approach for the realization of large area strain sensors with high sensitivity and low manufacturing costs. A polymer-based nanocomposite with carbon nanomaterials as conductive filler can be deposited on a flexible substrate of choice and this leads to mechanically flexible layers. Such sensors allow the strain measurement for both integral measurement on a certain surface and local measurement at a certain position depending on the sensor geometry. Strain sensors based on carbon nanostructures can overcome several limitations of conventional strain sensors, e.g., sensitivity, adjustable measurement range and integral measurement on big surfaces. The novel technology allows realizing strain sensors which can be easily integrated even as buried layers in material systems. In this review paper, we discuss the dependence of strain sensitivity on different experimental parameters such as composition of the carbon nanomaterial/polymer layer, type of polymer, fabrication process and processing parameters. The insights about the relationship between film parameters and electromechanical properties can be used to improve the design and fabrication of CNT strain sensors.

A mouse model that mediates temporal, specific, and efficient myocardial deletion with Cre-LoxP technology will be a valuable tool to determine the function of genes during heart formation. Mhy6 encodes a cardiac muscle specific protein: alpha-myosin heavy chain. Here, we generated a new Myh6-MerCreMer (Myh6(MerCreMer/+)) inducible Cre knock-in mouse by inserting a MerCreMer cassette into the Myh6 start codon. By crossing knock-in mice with Rosa26 reporter lines, we found the Myh6(MerCreMer/+) mice mediate complete Cre-LoxP recombination in cardiomyocytes after tamoxifen induction. X-gal staining and immunohistochemistry analysis revealed that Myh6-driven Cre recombinase was specifically activated in cardiomyocytes at embryonic and adult stages. Furthermore, echocardiography showed that Myh6(MerCreMer/+) mice maintained normal cardiac structure and function before and after tamoxifen administration. These results suggest that the new Myh6(MerCreMer/+) mouse can serve as a robust tool to dissect the roles of genes in heart development and function. Additionally, myocardial progeny during heart development and after cardiac injury can be traced using this mouse line.